Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity studies were conducted through oral route in rats, dogs and monkeys, through dermal route in rats and through subcutaneous route in monkeys. These studies were conducted with the dissociation products MEA and LAS as well as with the read-across substance C10-14LAS.

Considering the entire dataset, the highest NOAEL for systemic toxicity below the lowest LOAEL (115 mg/kg bw/day from the 26 weeks dietary study with C10-14 LAS in rats, Yoneyama, 1972, based on histopathological changes in liver and kidney) is 85 mg/kw bw/day from the 9 months drinking water study in rats with C10-14LAS (Yoneyama, 1976). In this drinking water study, the LOAEL was slightly higher at 145 mg/kg bw/day and based on liver and kidney enzyme changes. The consistency in the adverse effects seen in liver and kidneys in these studies (i.e. Yoneyama, 1976 and Yoneyama, 1972) supports the selection of 85 mg/kg bw/day as the appropriate NOAEL. Further, the selection of 85 mg/kg bw/day as the appropriate NOAEL is also supported by the higher NOAEL of 150 mg/kg bw/day observed in the 26 weeks oral gavage study in rats with C10 -13LAS (Ito, 1978b). Therefore, based on the entire data set, the point of departure taken forward for risk assessment purposes is the NOAEL of 85 mg/kg bw/day.

The registered substance is an MEA-LAS salt in a 1:1 molar ratio. Based on the molecular weight this corresponds to an MEA percent weight of 16% (w/w). The relevant NOAEL of LAS for risk assessment is 85 mg/kg bw/day while the repeat dose toxicity NOAEL for MEA is higher (300 mg/kg bw/day). In other words, the effects in an MEA-LAS repeat dose study are expected to be driven first by LAS, and at higher level also by MEA. At dose levels of 85 -100 mg/kg bw/day MEA-LAS (i.e., in the range of the NOAEL for LAS), the amount dosed of MEA would be 13.6 -16 mg/kg bw/day, which is well below the anticipated NOAEL for MEA (300 mg/kg bw/day). A repeat dose toxicity study on MEA-LAS is expected to deliver doses of MEA around one order of magnitude lower than those needed to observe MEA-effects.

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1972
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Principles of method if other than guideline:
The purpose of this study is to determine the sub chronic toxicity of C10 - C14 linear alkylbenzene sulfonic acid sodium salt (LAS) in Wistar SLC rats. Male and female rats were maintained on diets of 0, 0.07, 0.2, 0.6 and 1.8% LAS (equivalent to 0, 40, 115, 340, 1030 mg/kg bw/day) for 26 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, blood was collected for estimation of hematological and clinical chemistry parameters. Urinalysis was also performed 2 weeks before necropsy. All the surviving animals were humanely euthanized and gross necropsy, organs weights and histopathology were performed.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: Wistar SLC
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female were obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals
- Age at study initiation: Approximately 4 weeks
- Weight at study initiation: 80 - 90 g (male rats), 65 - 80 g (female rats)
- Fasting period before study: No
- Housing: 5 animals were housed per stainless steel cage.
- Diet: FII food (from Funabashi Farm); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 55 - 65%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: From: May 30, 1972 To: Nov. 27, 1972
Route of administration:
oral: feed
Details on route of administration:
C10 - C14 LAS was administered to animals by mixing 0.07, 0.2, 0.6 and 1.8% with pulverized FII feed, the powders were shaped into solid form before being freely provided for feeding along with tap water. As control, solid FII feed was provided to the animals.
Vehicle:
other: C10 - C14 LAS was administered in diet
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
26 weeks
Frequency of treatment:
Continuous in diet (ad libitum)
Dose / conc.:
0.07 other: %
Remarks:
in diet
Dose / conc.:
0.2 other: %
Remarks:
in diet
Dose / conc.:
0.6 other: %
Remarks:
in diet
Dose / conc.:
1.8 other: %
Remarks:
in diet
Dose / conc.:
40 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.07% dose level)
Dose / conc.:
115 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.2% dose level)
Dose / conc.:
340 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.6% dose level)
Dose / conc.:
1 030 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 1.8% dose level)
No. of animals per sex per dose:
10 animals/sex/dose group
Control animals:
yes, plain diet
Positive control:
No
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for examinations: Weekly

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: Weekly

OPHTHALMOSCOPIC EXAMINATION: No

HEMATOLOGY: Yes
- Time schedule for collection of blood: After 26 weeks, blood was collected from each animal prior to autopsy (for hemoglobin, hematocrit, red blood cells, white blood fractions and platelets) and during euthanasia (erythrocyte membrane resistance)
- Anesthetic used for blood collection: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red blood cells, white blood fractions and platelets, erythrocyte membrane resistance and blood coagulation time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 26 weeks, blood was collected from each animal after euthanasia.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Total proteins, GOT activity, ALP activity, LAP activity, urea nitrogen, A/G ratio and bilirubin

URINALYSIS: Yes
- Time schedule: 2 weeks prior to autopsy, urine isolated and collected from waste was analyzed.
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Proteins, glucose, ketone bodies, occult blood, bilirubin and urobilinogen

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Gross findings were obtained after euthanizing animals and macroscopic findings were observed.

ORGAN WEIGHTS: Brain, pituitary, thyroid, thymus, heart, lungs, liver, kidneys, spleen, adrenal gland, prostate, testes, uterus, ovaries and appendix

HISTOPATHOLOGY: Yes
Collected tissues along with stomach, large and small intestines, pancreas, epididymis, skin and bones) were weighed, examined and subjected to microscopy through H.E., PAS, lipid and silver staining.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Severe diarrhea was observed in the high concentration group (1.8%).
Mortality:
mortality observed, treatment-related
Description (incidence):
In 24th week, 1 male animal from the 1.8% dose group died.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was significant inhibition of weight gain in both males and females of 1.8% dose group.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption was low in the 1.8% dose group relative to the control.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
Water consumption was slightly low in animals of 1.8% dose group relative to the control
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
In females of 1.8% dose group, a significant decrease in hematocrit and hemoglobin and significant increase in the erythrocyte membrane resistance was observed.. For both males and female species, there was increase in white blood cell count at 1.8% dose.

Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
There was dose dependent significant increase in ALP levels (at mid and high dose) and significant decrease in total protein (at high dose).
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
1.8% dose group: Significant increase in relative weights of liver, suprarenal gland, testis, brain and caecum were observed in males and relative weights of heart, thyroid, liver, uterus, brain and caecum of females.
0.6% dose group: Significant increase in caecum relative weights in males and hypophysis in females.
0.2% dose group: Caecum relative weights were significantly increased in males and kidney relative weights were significantly decreased in females.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Color of the liver in 2 males and 5 females from the 1.8% group was lighter than normal and seemed to have a yellowish white color mixed to it. In addition, lung tumors were also seen in one female rat from the 0.2% group.

Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
The main findings were concerned with kidneys, liver and intestinal tract.
Kidney damage was seen at and above 0.2% dose groups. Mild hepatocyte changes were observed in animals of 1.8% dose group and microscopic changes in intestinal tract were observed in female rats of the 1.8 and 0.6% dose groups.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
CLINICAL SIGNS:
- Severe diarrhea was observed in animals in the high concentration groups (1.8%) immediately after starting the study, and although the diarrhea gradually became milder after about 1 week, the animal s did not recover fully by the end of the study. The animals moved slowly, and their lower abdomen was dirty.

MORTALITY: During week 24, 1 male animal from 1.8% dose group died.
BODY WEIGHT AND WEIGHT CHANGES: There was significant inhibition of weight gain in both males and females of 1.8% dose group, while the 0.6% dose groups showed slight but not significant trends to suppress weight gain.

FOOD CONSUMPTION AND COMPOUND INTAKE: Food consumption was low in the 1.8% dose group relative to the control.

WATER CONSUMPTION AND COMPOUND INTAKE: Water consumption was slightly low in the
1.8% dose group.

HEMATOLOGY:
- A significant decrease in hematocrit and hemoglobin, a significant increase in the erythrocyte membrane resistance was observed among female animals in the 1.8% dose groups.
- Male and female rats in 1.8% dose group showed increase in white blood cell count, increase in neutrophil fraction out of the white blood cells and a decrease in lymphocytes.

CLINICAL BIOCHEMISTRY: There was dose dependent increase in ALP levels (at 0.6 and 1.8% dose groups) and decrease in total protein (at 1.8% dose group)

URINALYSIS: No effects were observed.

ORGAN WEIGHT:
- In highest concentration group (1.8%), significant increase in relative weights of liver, suprarenal gland, testis, brain and caecum were observed in males and relative weights of heart, thyroid, liver, uterus, brain and caecum of females.In 0.6% dose group, there was significant increase in caecum relative weights in males and hypophysis in females.

In 0.2% dose group, caecum relative weights were significantly increased in males and kidney relative weights were significantly decreased in females.

GROSS PATHOLOGY
- The color of the liver in 2 males and 5 females from the 1.8% dose group was lighter than normal and seemed to have a yellowish white color mixed to it.
- In addition, lung tumors were seen in one female rat from the 0.2% dose group.

- Autopsy of one male rat from the LAS-1.8% dose group that died during the study had significant s welling of the abdomen. This was due to the swelling of the stomach and small intestines. The contents from the stomach to the first part of the jejunum was a blackish purple liquid. From there on, the ileum was filled with a semi-transparent, viscous, gelatinous substance.

HISTOPATHOLOGY

The main findings were concerned with kidneys, liver and intestinal tract. The histological findings concerning the kidneys had differences in the extent and frequency of changes between the dosage and between male and female rats.

Kidneys:
- Chronic appearance of glomeruli indicating moderate level of atrophy (male/females of 0.6% and 1.8% treated groups)
- Glomeruli contributed to overall swelling through dilation of vessel lumen and interstitial swelling (significant in male/female rats of 0.2 and 0.07% dose groups. They were not very significant in the 1.8 and 0.6% dose groups)
- The glomerular interstitum had thawn mildly but chronically (present in control group but was significant for male/female rats in 1.8 and 0.6% groups)
- Cloudy swelling/vacuolar degeneration of proximal renal tubular epithelial cells were seen (mildly present in control group and slightly significant in LAS treated groups).
- Small, yellowish brown blobs the size of nuclei was observed in the proximal renal tubular epithelial cells (significantly in male/females of 1.8% dose group)
- Blue lead color exhibited on the lumen of the Henle loop through H.E. staining, while substances giving a strong positive PAS response were forming layers in circular forms or were present in small pieces
-Lumen dilation of the paroximal renal tube and flattening of epithelial cells (significantly in males of 0.2% dose group, no effect observed in females and control groups)
- The renal tubule had collapsed (significantly in male/females of 1.8% dose group)
- Swelling of surrounding connective tissue was observed in the interstitium (observed in control group and significantly in males of LAS-0.2% dose group and both male and female rats of 0.6% dose group)

- Various changes to the kidneys as seen in 0.2% to 0.07% dose groups differ in their extent and frequency of incidence, but these changes were also seen in the control group, so they were not considered changes that lead to permanent damages like collapse of renal tubule or cyst formation.

Liver: Disappearance of basophilic substances in the hepatocytes and eosin thickening of hepatocytes were observed mildly in male and female rats of the 1.8% dose group.

Intestinal tract: Decrease in height of epithelial cells in the colonic mucosa and disappearance of vacuoles from goblet cells were observed mildly in female rats of the 1.8 and 0.6% dose groups.

Further details on are provided in "Any other information on results inc.tables"
Key result
Dose descriptor:
NOAEL
Effect level:
40 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: Suppression of weight gain, increase in appendix weight, alteration in hematological and clinical biochemistry parameters, and/ or tissue damages to the large intestines, liver and kidneys in dose levels at ≥ 115 mg/kg bw/day (0.2% diet).
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
115 mg/kg bw/day (nominal)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1:  Body weight gain, food, detergent, water intake and feces excretion in males (Yoneyama et al., 1972)

Group  No. of Rats Initial body wt (g) (mean±S.E) Final body wt (g) (mean±S.E) Body weight gain (1) (%) (mean±S.E) Food (g/rat/day) Detergent water (g/rat/day) Feces (g/rat/day)
mg/rat/day mg/kg/day
Control 10 85±0.871 387±9.509 452.1±9.841 14.17 - - 16.9 2.31
1.80% 10-9 85.2±0.553 290.00±5.206 341.09±5.136** 9.84 177.12 814.8 13.29 1.78
0.60% 10 86.40±1.543 365.80±8.517 426.23±8.893 13.84 83.04 322.3 16.28 2.23
0.20% 10 84.50±0.957 383.40±6.715 453.92±7.603 13.7 27.4 101.5 18.18 2.28
0.07% 10 80.9±1.923 387.10±5.930 480.74±13.113 14.48 10.14 37.2 16.91 2.24

Table 2:  Body weight gain, food, detergent, water intake and feces excretion in females (Yoneyama et al., 1972)

Group  No. of Rats Initial body wt (g) (mean±S.E) Final body wt (g) (mean±S.E) Body weight gain (%) (1) (mean±S.E) Food (g/rat/day) Detergent water (g/rat/day) Feces (g/rat/day)
mg/rat/day mg/kg/day
Control 10 71.80±0.512 202.50±2.325 282.24±4.422 9.35 - - 17.48 1.53
1.80% 10 74.30±0.943 158.80±3.411 214.02±5.288** 7.01 126.18 913 12.14 1.23
0.60% 10 74.10±0.673 195.80±4.149 264.25±5.207* 9.04 54.24 332.2 13.99 1.53
0.20% 10 74.30±1.116 201.10±3.334 271.05±5.314 8.9 17.8 108.7 17.69 1.39
0.07% 10 76.00±2.683 203.20±7.172 269.35±10.255 9.35 6.55 39.4 19.56 1.63

where (1) refers to Final BW/Initial BW-1) into100

*P<0.05

**P<0.01

Table 3 : Hematology-I in males and females rats (Yoneyama et al., 1972)

Group No. of Rats RBC*104/mm3  Ht (%) Hb (g/dl) MCH (rr) MCV (µ8) MCC%
Male
Control 10 910±13 53.1±0.4 19.8±0.3 21.8±0.4 58.4±1.0 37.4±0.6
1.80% 9 924±271) 54.6±0.4 20.6±0.5 22.4±0.8 59.3±1.3 37.7±0.8
0.60% 10 890±14 54.0±0.4 19.8±0.1 22.3±0.3 60.7±0.9 36.7±0.1
0.20% 10 914±25 53.3±0.3 19.5±0.1 21.5±0.6 58.6±1.6 36.7±0.2
0.07% 10 925±20 53.3±0.4 19.6±0.1 21.2±0.5 57.8±1.2 36.7±0.2
Female
Control 10 838±22 52.1±0.5 19.9±0.5 23.9±0.9 62.5±1.6 38.2±1.0
1.80% 10 785±14 49.5±0.3** 18.0±0.1** 23.0±0.4 63.2±0.9 36.4±0.1
0.60% 10 795±17 50.9±0.3 19.0±0.1 24.0±0.5 64.2±1.1 37.4±0.2
0.20% 10 778±16 51.1±0.2 19.1±0.1 24.7±0.4 65.9±1.3 37.5±0.2
0.07% 10 806±18 50.6±0.4 18.9±0.1 23.6±0.6 63.0±1.5 37.4±0.3

Table 4 : Hematology-II in males and females rats (Yoneyama et al., 1972)

Group No. of Rats Thrombocyte*104(A) Osmotic fragility (A) Whole blood clotting time (min)
Beginning Ending
Male
Control 10 70.5±9 (8) 213.6±9.4 (9) 1.06±0.13 2.12±0.26
1.80% 9 70.2±4.9 (8) 199.9±6.6 (9) 1.14±0.11 2.32±0.27
0.60% 10 71.6±8.4 211.4±7.9 1.42±0.23 2.71±0.29
0.20% 10 64.4±5.5(9) 228.1±6.5(9) 1.34±0.19 2.50±0.35
0.07% 10 80.1±13.2 225.1±10.5 1.40±0.19 2.52±0.33
Female
Control 10 52.0±3.8 (9) 240.4±7.5 (8) 1.29±0.29 2.95±0.30
1.80% 10 58.4±4.5 (9) 181.8±6.5(8)* 1.37±0.14 2.57±0.19
0.60% 10 54.8±3.3 (9) 226.0±5.9 (9) 1.24±0.17 2.81±0.22
0.20% 10 59.8±5.9 (9) 245.4±9.3(9) 1.26±0.19 2.92±0.22
0.07% 10 63.3±5.3 (8) 248.4±10.2 1.22±0.21 3.30±0.41

*P<0.05

**P<0.01

(A): No of rats

1) (mean±SE)

Table 5: Percent differential of Leucocytes (Yoneyama et al., 1972)

Group No. of Rats Neutro
Lympho Mono Eosino
Male
Control 10 9.8±0. 88 (1) 89.08±0.954 0.28±0.102 0.83±0.203
1.80% 9 12.09±0.698* 87.22±0.624 0.09±0.061 0.58±0.164
0.60% 10 10.77±1.017 87.82±0.860 0.26±0.094 0.60±0.158
0.20% 10 10.34±0.521 88.22±0.625 0.40±0.102 0.62±0.128
0.07% 10 9.41±1.074 89.35 ± 0.998 0.50± 0.088 0.73±0.125
Female
Control 10 8.24±0.56 90.85±0.685 0.18±0.073 0.72±0.150
1.80% 10 8.67±0.747 90.61 ± 0.762 0.28±0.076 0.42±0.108
0.60% 10 10.21±1 .054 88.49±1.065 0.32±0.071 0.96±0.143
0.20% 10 8.75±0.754 90.15±0.781 0.38±0.096 0.70±0.146
0.07% 10 8.65±0.752 90.39 ± 0.771 0.40±0.099 0.54±0.108

Table 6: Counts differential of leucocytes.102/mm3 (Yoneyama et al., 1972)

Group No. of Rats WBC Neutro Lympho Mono Eosino
Male
Control 10 88.12±3.686 (1) 8.49±0.678 78.65±3.779 0.24±0.087 0.67±0.148
1.80% 9 96.16±5.726 11.78±1.113 83.76±4.805 0.08 ±0.057 0.54±0.142
0.60% 10 91.47±5.430 9.96±1.154 80.27±4.686 0.25±0.088 0.58±0.142
0.20% 10 90.55±6.435 9.61±1.166 79.64±5.196 0.36 ±0.089 0.60±0.155
0.07% 10 96.15±3.329 9.08±1.077 85.85±2.913 0.50±0.093 0.71±0.135
Female
Control 10 92.00±3.802 7.59±0.579 83.57±3.517 0.16±0.066 0.67±0.139
1.80% 10 109.30±3.831** 9.34±0.664 95.03 ± 7.208 0.31 ± 0.085 0.47±0.120
0.60% 10 87.90±3.631 8.87±0.847 77.89±3.679 0.26±0.058 0.85±0.133
0.20% 10 83.15±4.350 7.38±0.637 76.67 ± 5.063 0.32±0.078 0.61 ± 0.138
0.07% 10 94.40±5.110 8.36±1.176 85.13±4.180 0.37±0.091 0.51 ± 0.101

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 7: Serum Chemistry (Yoneyama et al., 1972)

Group No. of Rats ALP
K-A Unit/dl
LAP
G-R Unit
GOT
karmen unit/ml
UN
mg/dl
A/G ratio Total protein
g/dl
Bililubin
                (Indirect)
mg/dl
(Direct)
mg/dl
Total
mg/dl
Male      
Control 10 24.07±1.1091) 185.81± 9.586 132.75± 6.357 16.92±1.102 1.42±0.082 7.36±0.071 0.20±0.050 0.11±0.036 0.29±0.044
1.80% 9 44.96±4.01** 179.80± 9.74 139.12± 9.397 16.30±0.634 1.59±0.129 6.55±0.191** 0.40±0.250 0.12±0.027 0.48±0.268
0.60% 10 29.22±1.093** 180.54± 7.897 133.50± 5.559 15.76±0.611 1.46±0.096 7.20±0.084 0.30±0.201 0.32±0.211 0.56±0.415
0.20% 10 24.34±1.173 181.23± 9.637 140.25±14.732 13.04±1.224* 1.47±0.091 7.20±0.059 0.41±0.231 0.31±0.188 0.68±0.422
0.07% 10 25.48±0.816 178.99± 9.4271 144.60±11.501 15.36±1.221 1.38±0.082 7.26±0.118 0.35±0.201 0.34±0.197 0.64±0.401
Female      
Control 10 24.41±1.380 198.54± 8.899 156.30±12.716 17.52±1.733 1.53±0.062 7.80±0.122 0.12±0.050 0.14±0.054 0.23±0.054
1.80% 10 61.41±6.124** 208.72± 9.097 154.60±8.297 19.00±1.043 1.76±0.121 6.42±0.127** 0.12±0.025 (9) 0.12±0.033 (9) 0.25±0.054 (9)
0.60% 10 31.80±1.599** 187.58± 10.642 139.50±3.024 17.15±2.020 1.70±0.097** 7.22±0.186* 0.14±0.036 0.12±0.038 0.27±0.062
0.20% 10 27.23±1.049 193.65± 7.686 146.55±9.144 14.14±1.404 1.48±0.164 7.38±0.184 0.20±0.044 (9) 0.11±0.023 (9) 0.31±0.052 (9)
0.07% 10 25.77±1.237 192.23± 8.056 128.25±5.432 15.09±1.245 1.72±0.067 7.68±0.163 0.15±0.059 (9) 0.10±0.018 (9) 0.24±0.06 (9)

where:

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 8 : Urinalysis in female and male rats (Yoneyama et al., 1972)

Group No. of Rats pH Protein Glucose Keton Blood Bilirubin Urobilin
    8 7 6 5 - ± + ++ +++ - + - ± + - ± + - + 0.1* 1
Male
Control 10 1 4 5 - - - 7 2 1 10 - 7 3 - 10 - - 10 - 10 -
1.80% 9 - - 9 - - - 2 3 4 9 - 2 7 - 9 - - 9 - 9 -
0.60% 10 - 5 5 - - 2 5 2 1 10 - 5 5 - 10 - - 10 - 10 -
0.20% 10 - 2 8 - - 4 3 3   10 - 6 4 - 10 - - 10 - 10 -
0.07% 10 - 4 6 - 1 - 6 1 2 10 - 8 2 - 10 - - 10 - 10 -
Female
Control 10 2 2 6 - 2 2 5 1 - 10 - 5 5 - 10 - - 10 - 10 -
1.80% 10 1 - 9 - 3 3 4 - - 10 - 4 5 - 10 - - 10 - 9 -
0.60% 10 - 5 5 - 2 3 2 3 - 10 - 7 3 - 10 - - 10 - 10 -
0.20% 10 - 3 7 - 2 2 5 1 - 10 - 4 6 - 10 - - 10 - 10 -
0.07% 10 - 3 7 - 3 3 4 - - 10 - 7 3 - 10 - - 10 - 9 -

*Ehrlich units

Table 9: Organ Weights (Male-1) (Yoneyama et al., 1972)

Group  No. of rats Heart
[Actual weight (mg)/weight (mg)/100 g]
Spleen
[Actual weight (mg)/weight (mg)/100 g]
Thymus
[Actual weight (mg)/weight (mg)/100 g]
Liver
[Actual weight (g)/weight (g)/100 g]
Lung
[Actual weight (mg)/weight (mg)/100 g]
Kidneys
[Actual weight (mg)/weight (mg)/100 g]
Hypophysis
[Actual weight (mg)/weight (mg)/100 g]
  R L
Control 10 930±13/241± 31) 614±14/158±2 129±5.7/33±0.9 12.0±0.38/3.1±0.04 1211±44/314±13 1105±33/285± 6 1126±33/219± 6 10.0±0.73/2.6±0.21
1.80% 9 694±11**/239±3 460±11**/158±2 102±6.3**/35±1.7 10.3±0.28**/3.5±0.08** 971±35**/324±13 836±19**/288±3 852±22**/294±5 8.8±0.42/3.0±0.15
0.60% 10 878±18*/235± 4 572±15*/153± 4 122±8.8*/32± 2.3 12.0±0.31/3.2±0.07 1080±21/289±6 1045±31/280±9 1049±34/281±9 9.4±0.30/2.5±0.08
0.20% 10 918±16/239± 2 644±27/168±7 130±9.0/33±2.1 12.3±0.42/3.2±0.12 1253±63/326±14 1083±34/282±7 1071±28/279±4 8.9±0.48/2.3±0.13
0.07% 10 932±16/240± 2 641±12/165± 2 129±10.8/33± 2.5 12.0±0.31/3.1±0.05 1285±47/332±12 1074±25/277±5 1083±52/279±12 9.3±0.50/2.4±0.13

Table 10: Organ Weights (Male-2) (Yoneyama et al., 1972)

Group  No. of rats Thyroid glands
[Actual weight (mg)/weight (mg)/100 g]
Supranal gland
[Actual weight (mg)/weight (mg)/100 g]
Testis
[Actual weight (mg)/weight (mg)/100 g]
Prostate glands
[Actual weight (mg)/weight (mg)/100 g]
Brain
[Actual weight (mg)/weight (mg)/100 g]
Caecum
[Actual weight (mg)/weight (mg)/100 g]
  R L R L
Control 10 14.4±0.4211)/3.7±0.16 18.0±0.61/4.6±0.18 19.3±0.44/5.0±0.17 1592±20/412± 7 1622±21/420± 6 252±16/65±4.8 1983±12/515±11 837±34/216± 8
1.80% 9 11.5±0.76**/3.9±0.24 15.5±0.44**/5.3±0.17* 17.1±0.65*/5.9±0.27* 1513±23*/522±7** 1501±36**/518±13** 231±21/79±7.0   1960±22/677±11 ** 1484±30**/512±12**
0.60% 10 13.2±0.69/3.5±0.22 17.5±0.76/4.6±0.20 18.2±0.69/4.8±0.19 1576±27/422± 11 1612±36/432±11 231±21/61±5.7 1969±18*/528±12 939±41/251±11*
0.20% 10 13.3±0.57/3.4±0.11 17.3±0.61/4.5±0.17 18.6±0.84/4.8±0.23 1564±22/408± 5 1621±20/423± 4 298±21/78±5.9 2020±11/528±8 971±35*/253 ± 7**
0.07% 10 13.6±0.54/3.5±0.13 16.8±0.85/4.3± 0.22 18.7±0.44/4.8± 0.11 1643±14/425± 6 1676±15/434± 7 318±21*/82±6.1 1918±106/494±25 938±38/242± 9

where,

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 11: Organ Weights (Female-1) (Yoneyama et al., 1972)

Group  No. of rats Heart
[Actual weight (mg)/weight (mg)/100 g]
Spleen
[Actual weight (mg)/weight (mg)/100 g]
Thymus
[Actual weight (mg)/weight (mg)/100 g]
Liver
[Actual weight (g)/weight (g)/100 g]
Lung
[Actual weight (mg)/weight (mg)/100 g]
Kidneys
[Actual weight (mg)/weight (mg)/100 g]
Hypophysis
[Actual weight (mg)/weight (mg)/100 g]
R L
Control 10 596±1001)/265± 6 405± 5/200±4 134±7.7/66±3.9 6.5±0.12/3.2±0.08 822±19/409±11 720±22/ 359±12 738±18/395±11 13.9±0.76/6.8±0.39
1.80% 10 426± 11**/269±7* 294± 10**/185± 6 102±6.8**/64±4.1 6.3±0.24/3.9±0.12** 673±20**/425±16 546±16**/345±13 563±24**/357±21 12.3±1.08/7.7±.63
0.60% 10 559± 11*/286± 5 387 ± 7/198±5 125±5.8/63±2.2 6.5±0.22/3.3±0.10 793±14/406±8 664±16/340± 9 715±32/ 365±14 16.2±0.94/8.3±0.46*
0.20% 10 574±13/258± 5 405 ±8/201 ± 4 131±4.6/65±2.6 6.5±0.18/3.2±0.07 802±26/398±10 654 ±20*/325± 7 665±28*/330±11* 15.0±1.10/7.4±0.53
0.07% 10 596±16/294± 3 408±13/201± 4 132±2.8/65±2.3 6.7±0.30/3.3±0.05 801±19/396±11 717±23/355±13 738±33/364±13 16.3±0.94/8.0±0.40

Table 12: Organ Weights (Female-2) (Yoneyama et al., 1972)

Group  No. of rats Thyroid glands
[Actual weight (mg)/weight (mg)/100 g]
Supranal gland
[Actual weight (mg)/weight (mg)/100 g]
Ovary
[Actual weight (mg)/weight (mg)/100 g]
Uterus
[Actual weight (mg)/weight (mg)/100 g]
Brain
[Actual weight (mg)/weight (mg)/100 g]
Caecum
[Actual weight (mg)/weight (mg)/100 g]
  R L R L
Control 10 11.2±0.3211)/ 5.5±0.17 21.9±0.99/10.8±0.64 23.2±1.24/11.4±0.94 23.3±1.0/11.5±0.5 24.4±1.1/12.0±0.5 638±27/315±13 1832±16/906±14 681±18/337±11
1.80% 10 10.9±0.73/6.9±0.53* 15.6±0.81**/9.8±0.52 16.8±0.84**/10.6±0.55 17.6±1.4**/11.1±0.9 18.2±1.8**/11.5±1.1 292±18**/185±12** 1780±15*/1126±28 929 ± 26**/586±14**
0.60% 10 13.9±1.30/7.1±0.70 22.9±0.56/11.7±0.36 24.0±0.61/12.3±0.44 22.8±1.2/11.7±0.7 24.5±1.0/12.6±0.6 618±32/315±15 1840±14/943±20 697±21/356± 7
0.20% 10 12.2±0.96/6.0±0.46 21.7±1.11/10.7±0.49 22.2±0.92/11.0±0.43 23.6±1.0/11.7±0.4 23.3±1.2/11.5±0.5 594±31/296±17 1837±18/915±13 688±26/343±14
0.07% 10 12.9±0.93/6.4±0.55 22.6±0.95/11.1±0.35 24.1±0.98/11.8±0.33 23.0±1.9/11.2±0.6 24.6±1.5/12.0±0.5 592±18/292± 8 1827±18/908±30 753±25*/373±13

where,

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 13: Summary of microscopic  changes in kindey (Yoneyama et al., 1972)

Microscopic findings # Male Female
C LAS C LAS
0.07% 0.2% 0.6% 1.8% 0.07% 0.2% 0.6% 1.8%
Swelling in glomerulus S E
M O
S L
0
2 (10)
2
0
5 (10)
4
1
6 (10)
3
0
7 (10)
2
0
5 (9)
4
0
2 (10)
3
1
5 (10)
4
1
5 (10)
4
0
7 (10)
3
0
5 (10)
5
Histolysis in glomerulus S E
M O
S L
0
2
2
0
7
2
1
5
4
1
5
4
1
5
3
0
3
1
0
5
5
0
9
4
1
6
3
1
5
4
Atrophy in glomerulus S E
M O
S L
0
1
1
0
5
5
1
6
2
2
7
1
3
6
0
0
1
1
0
4
5
0
6
3
1
7
2
4
5
1
Cloudy swelling in epithelium of proximal tubules S E
M O
S L
0
0
4
0
1
5
0
6
2
0
8
2
0
7
2
0
0
3
0
0
4
0
3
3
0
3
3
0
4
5
Vacuolation in epithelium of proximal tubules S E
M O
S L
0
0
4
0
6
4
0
3
7
0
4
6
0
5
4
0
1
5
0
7
3
0
6
4
0
7
3
0
7
3
Lipofuscin in epithelium of proximal tubules S E
M O
S L
0
0
0
0
0
0
0
0
5
0
0
5
2
7
0
0
0
0
0
0
0
0
0
1
0
0
4
5
5
0
PAS-positive materies in loop of Henle S E
M O
S L
          0
0
4
0
3
7
0
3
6
0
3
6
0
0
3
Dilatation in lumina of distal tubules S E
M O
S L
0
0
2
0
4
6
3
3
4
2
4
4
0
5
4
0
0
0
0
0
5
0
3
3
0
5
5
0
3
5
Decrease in cell height of epithelium in distal tubules S E
M O
S L
0
0
1
0
0
6
2
3
5
2
3
4
0
4
5
0
0
0
0
0
3
0
0
5
0
3
5
0
3
5
Destruction in urinary tubules S E
M O
S L
0
0
0
0
0
0
0
0
2
0
2
3
0
3
4
0
0
0
0
0
2
0
0
3
0
0
5
0
4
5
edema in stroma S E
M O
S L
0
1
2
1
5
4
1
6
3
1
3
5
0
4
5
0
1
1
1
4
5
1
5
4
1
6
3
0
2
7
Swelling in perivascular connective tissues S E
M O
S L
0
1
1
1
4
4
1
3
6
0
5
5
0
3
4
0
1
1
0
4
5
0
5
5
0
7
3
0
1
6
Proliferation in interstitial connective tissues S E
M O
S L
0
0
1
0
0
5
0
0
5
0
1
4
0
2
6
0
0
2
0
0
2
0
0
5
0
0
6
0
0
6

where, SE: Severe

SL: Slight

MO: Moderate

(A) No. of rats

Table 14:  Summary of microscopic changes in liver and large intestine (Yoneyama et al., 1972)

Microscopic findings # Male Female
C LAS C LAS
0.07% 0.2% 0.6% 1.8% 0.07% 0.2% 0.6% 1.8%
Decrease in basoplam in liver cells S E
M O
S L
0
0 (10)
0
0
0 (10)
0
0
0 (10)
1
0
0 (10)
1
0
0 (9)
6
0
0 (10)
0
0
0 (10)
0
0
0 (10)
0
0
0 (10)
3
0
1 (10)
6
Increase in eosinophilia in liver cells S E
M O
S L
0
0
1
0
0
0
0
0
0
0
0
0
0
0
6
0
0
1
0
0
0
0
0
0
0
0
2
0
2
7
Swelling in liver cells S E
M O
S L
0
0
0
0
0
0
0
0
0
0
3
3
0
3
3
0
0
0
0
0
0
0
1
3
0
1
4
0
2
6
Rarefaction in cytoplasm of liver cells S E
M O
S L
0
0
3
0
0
8
0
1
6
0
1
6
0
1
6
0
0
2
0
0
6
0
1
7
0
1
7
0
1
7
Decrease in cell height of epithelium in large instestine S E
M O
S L
0
0
0
0
0
0
0
0
2
0
0
4
0
0
5
0
0
0
0
0
0
0
0
3
0
1
4
0
1
4

where, SE: Severe

SL: Slight

MO: Moderate

(A) No. of rats

Conclusions:
Administration of linear alkylbenzene sulfonic acid sodium salt (LAS) to Wistar SLC rats at dose levels of 0, 0.07, 0.2, 0.6 and 1.8% (equivalent to 0, 40, 115, 340, 1030 mg/kg bw/day) for 26 weeks revealed an NOAEL of 0.07% (40 mg/kg bw/day), based on tissue damage in caecum, kidney and liver, hematological and/or clinical chemistry parameters changes at higher dose levels (115 mg/kg bw/day and above).
Executive summary:

A sub-chronic repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in Wistar SLC rats. The test substance was administered to groups of 10 male and female rats/dose in the diet for 26 weeks at doses of 0, 40, 115, 340 and 1030 mg/kg/day. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, haematological, clinical chemistry and urine parameters were analysed. Gross pathological examinations were observed after necropsy and the major organs were subjected to histopathological examinations. Significant diarrhoea, suppressed growth, increased caecal weight and degeneration of renal tubes were observed in the highest dose group. Similar but less severe signs were seen in other doses with the exception of the lowest dose of 40 mg/kg/day, which showed no adverse effects related to exposure to the LAS. Under the study conditions, systemic toxicity NOAEL was determined to be 40 mg/kg/day, based on tissue damage in caecum, kidney and liver, haematological as well as clinical chemistry parameters changes at higher dose levels (Yoneyama, 1972). 

Endpoint:
sub-chronic toxicity: oral
Remarks:
other: two-generation reproductive toxicity study
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Qualifier:
according to
Guideline:
other: OECD Guideline 416 (Two-generation reproduction toxicity study)
Deviations:
yes
Remarks:
Food consumption was not determined between days 14 and 21 after parturition
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
other: Crl:WI (Han)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services GmbH, Germany
- Age at study initiation: (P) 16 days
- Weight at study initiation: (P) Males: 162.1 (142.5 – 186.5) g ; Females: 126.2 (110.6 – 145.1) g;
- Fasting period before study: none
- Housing: housed individually in type DK III stainless steel wire mesh cages
- Diet: ground Kliba maintenance diet mouse/rat “GLP” meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerland ad libitum
- Water: ad libitum
- Acclimation period: 16 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12


Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
The test substance (ethanolamine hydrochloride, EAH) was weighed and thoroughly mixed with a small amount of food. Then corresponding amounts of food, depending on the dose group, were added to this premix in order to obtain the desired concentrations. Mixing was carried out for about 10 minutes in a laboratory mixer. Test diets were prepared at intervals, which guaranteed that the test substance in the diet remained stable throughout the feeding period.

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability of EAH in the diet over 32 days at room temperature was investigated analytically before the beginning of the study. Homogeneity and concentration control analyses were carried out at the beginning and toward the end of the premating periods. At least one analysis of test substance preparations for female animals was carried out during the gestation and lactation periods.

The analyses were carried out at the Analytical Chemistry Laboratory of Experimental Toxicology and Ecology of BASF SE, Ludwigshafen, Germany.
Duration of treatment / exposure:
semichronic duration (> 75 days)
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
100, 300 and 1000 mg/kg bw/day
Basis:
nominal in diet
No. of animals per sex per dose:
25
Control animals:
yes, plain diet
Observations and examinations performed and frequency:
For parental animals:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily on working days and once daily on weekends

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: body weights of F0 and F1 parents were determined once weekly; during gestation and lactation F0 and F1 females were weighed on days 0, 7, 14 and 20 of gestation, and on days 1, 4, 7, 14 and 21 after birth.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Time schedule: once weekly (over a period of at least 6 days each) and weekly during gestation (days 0-7, 7-14, 14-20 post coitum; p.c.) and lactation periods (days 1-4, 4-7, 7-14 post partum; p.p.).

OTHER:
The F1 and F2 pups were sexed on the day of birth (day 0 p.p.) and weighed on days 1, 4, 7, 14, and 21 p.p. Their viability was recorded. At necropsy, all pups were examined macroscopically (including weight determinations of brain, spleen and thymus in one pup/sex/litter).

Serum concentrations of the test substance:
Blood samples were taken from all F0 and F1 parental animals of each sex and test group during week 10 of premating treatment and the plasma was analyzed for the concentration of Ethanolamine hydrochloride

Estrous cycle data were evaluated for F0 and F1 generation females over a three week period prior to mating until evidence of mating occurred. Moreover, the estrous stage of each female was determined on the day of scheduled sacrifice.

Parameters examined in [all/P/F1] male parental generations:
motility, sperm head count, morphology
Sacrifice and pathology:
For parental animals:
All F0 and F1 parental animals were sacrificed by decapitation under Isoflurane anesthesia. The exsanguinated animals were necropsied and assessed by gross pathology, special attention was given to the reproductive organs. The liver, kidneys, adrenal glands, testes, epididmides. Cauda epididymis, prostate, seminal vesicles, ovaries, uterus, spleen, brain, pituitary gland and thyroid glands (with parathyroids) were weighed and the vagina, cervix uterie, uterus, ovaries, oviducts, left testis, left epididymis, seminal vesicles, coagulation glands, prostate, pituitary gland, adrenal glands, liver, kidneys, spleen, brain, thyroids (with parathyroids)and all gross lesions were fixed in an appropriate fixative, histologically processed and examined by light microscopy. From both ovaries (”ovary 1” and “ovary 2”) of F1 female animals (control and top dose), five sections were taken from the proximal and the distal part of the ovaries, at least 100 µm apart from the inner third of the ovary. All ovarian sections were prepared and evaluated for numbers of primordial and growing follicles.
As soon as possible after termination, one portion of the liver (lobus medialis) of each 10 dams per group was sampled to be analyzed for choline concentration.
Statistics:
See below
Details on results:
Clinical examinations revealed no test substance-related adverse effects for F0 and F1 parental animals of low (100 mg/kg bw/dayand mid (300 mg/kg bw/daylevels. The test compound did adversely affect food consumption of the high dose F0 females (1000 mg/kg bw/day) during lactation. Also, the body weight gain and, for F0 generation, body weights of the high-dose dams were statistically significantly less compared to the control group during gestation, which was likely be secondary to an increased post-implantation loss in these
animals.

Estrous cycle data and sexual organ weights and morphology were comparable between the F0 and F1 dams of all test groups and the corresponding controls and ranged within the historical control data of the test facility.
In the top-dose F0 and F1 males the test substance administration led to a decrease of absolute and relative organ weights of cauda epididymidis and epididymides. Furthermore, prostate weight and the number of homogenization resistant caudal epididymal sperm was slightly, but significantly decreased in the F0 males. These findings were considered to be treatment-related effects, whereas histomorphological correlates were missing.

A statistically significant increase of absolute and relative kidney weights was noted in male and female F1 animals of the mid (300 mg/kg bw/day) and top-dose (1000 mg/kg bw/day) groups. Because no histomorphological correlate was detected, the treatment-related weight increase was considered to be of no toxicological concern. As compared to control animals, the kidneys of low-, mid-, and top-dose male and female animals revealed a low incidence of basophilic tubules in a slightly higher number of animals. The severity (minimal to slight) was comparable between controls and treated animals and a clear dose-response relationship was missing. Thus this finding was considered to have no toxicological relevance.
Dose descriptor:
NOAEL
Remarks:
parental
Effect level:
300 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: reduced food consumption and/or body weight gain, as well as organ weight changes unaccompanied by histopathological findings
Critical effects observed:
not specified

Test substance stability:

The stability of test substance in rat diet was demonstrated for a period of 32 days at room temperature in a different batch of comparable quality, which was not used for the study. The homogeneity of the mixtures was verified. The concentration control analyses of the samples taken revealed that the values were within a range of 90-110% of the nominal concentration in all analyses at all time points, with the exception of one concentration in the feed of the high-dose group (88%).

Plasma concentrations of 2 -aminoethanol were below 3 mg/kg for all control animals, <3 - 4 mg/kg for the low dose animals, 8 - 11 mg/kg for the mid dose animals and 60 – 81 mg/kg for the high dose animals.

Toxicokinetic dataof 2 -aminoethanol (calculated as 2 -aminoethanol hydrochloride)fromthis two-generation reproduction toxicity studyshow a dose dependency of the plasma levels of 2 -aminoethanol in the experimental animals and there with prove the bioavailability of 2 -aminoethanol hydrochloride in principle.

 

Tables

Mean test substance intake (mg/kg bw/d; minimum value / maximum value)

 

Test group 01
(100 mg/kg bw/d)

Test group 02
(300 mg/kg bw/d)

Test group 03
(1000 mg/kg bw/d)

F0 males

94.3 (72.4 / 102.5)

283.2 (218.4 / 309.4)

943.3 (716.7 / 1032.6)

F0 females (premating)

96.7 (80.5 / 100.7)

289.6 (241.2 / 304.9)

964.4 (792.4 / 1017.8)

F0 females
(F1 litter)
- gestation period
- lactation period*



103.5 (92.6 / 111.6)
99.2 (81.6 / 120.2)



315.2 (284.8 / 337.9)
306.7 (249.7 / 370.3)



1043.2 (989.4 / 1084.7)
866.0 (668.6 / 1053.9)

* = Days 1–14 p.p. only

Absolute organ weights (P-generation)

Compared to the controls (= 100%), the following values (in %) were significantly changed (printed in bold):

 

Male animals

Female animals

Group

01

100 mg/kg bw/d

02

300 mg/kg bw/d

03

1000 mg/kg bw/d

01

100 mg/kg bw/d

02

300 mg/kg bw/d

03

1000 mg/kg bw/d

Brain

99%

100%

97%*

 

 

 

Cauda epididymis

99%

102%

88%**

 

 

 

Epididymides

100%

101%

92%**

 

 

 

Prostate

92%

99%

86%**

 

 

 

Spleen

 

 

 

105%*

107%

97%

 

*: p≤0.05; **: p≤0.01

 

All other mean absolute weight parameters did not show significant differences compared to the control groups.

 

The decrease of absolute weights of cauda epididymis, epididymides, and prostate in male top-dose animals (1000 mg/kg bw/d) were considered as treatment-related effects.

 

The decrease of brain weights in top-dose males (1000 mg/kg bw/d) as well as the increase of spleen weights in low-dose females (100 mg/kg bw/d) was considered as incidental and not treatment-related due to a missing dose-response relationship.

Absolute organ weights (F1 generation)

Compared to the controls (= 100%), the following values (in %)were significantly changed (printed in bold):

 

 

Male animals

Female animals

Group

11

100 mg/kg bw/d

12

300 mg/kg bw/d

13

1000 mg/kg bw day

11

100 mg/kg bw/d

12

300 mg/kg bw/d

13

1000 mg/kg bw/d

Cauda epididymis

96%

99%

88%**

 

 

 

Epididymides

100%

101%

91%**

 

 

 

Kidneys

99%

106%*

111%**

103%

106%**

115%**

Spleen

99%

103%

92%*

 

 

 

Thyroid glands

106%

99%

109%*

110%

118%**

111%*

 

*: p≤0.05; **: p≤0.01

All other mean absolute weight parameters did not show significant differences compared to the control groups.

The decrease of absolute weights of cauda epididymis and epididymides in male top-dose animals (1000 mg/kg bw/d) were considered to be treatment-related.

 

The increase of absolute kidney weights of male and female animals in mid- (300 mg/kg bw/d) and top-dose (1000 mg/kg bw/d) groups, respectively, was statistically significant. Because no histomorphological correlate was detected, a treatment-related weight increase was less likely.

 

The decrease of spleen weights in top-dose males as well as the increase of thyroid glands in top-dose males and mid- and top-dose females, respectively, is considered incidental and not treatment-related due to a missing dose-response relationship.

Executive summary:

Effects of repeated exposure to monoethanolamine (MEA) were evaluated in the context of a two-generation reproductive toxicity study according to OECD Guideline 416 in rats. The test substance was administered to 25 rats/sex/dose via diet at doses of 0, 100, 300 and 1,000 mg/kg/day (nominal) through two successive generations. Clinical examination revealed no test substance-related adverse effects in F0 and F1 parental animals of low and mid doses. The test substance adversely affected food consumption at the high dose F0 females during lactation. Also, the body weight gain and, for F0 generation, body weights of the high-dose dams were statistically significantly less compared to the control group during gestation, which was likely be secondary to an increased post-implantation loss in these animals. Regarding the gross pathological examinations, the decrease of absolute weights of cauda epididymis, epididymis, and prostate in F0 and F1 male top-dose animals were considered as treatment-related effects. There were no other treatment-related systemic effects in any of the generations. Under the study conditions, the systemic toxicity NOAEL was determined to be 300 mg/kg/day (ACC and CEFIC, 2009). 

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Reason / purpose:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
This range finding repeated dose oral toxicity test was performed to establish suitable dose levels of test substance for a repeated dose oral toxicity study in rats. Test substance administered in the food to groups of ten rats for 90 days. The animals were examined for any abnormalities, if any and LD50 was calculated.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
not specified
Details on species / strain selection:
not specified
Sex:
not specified
Details on test animals and environmental conditions:
No details on test animals were provided.
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: not specified

- DIET PREPARATION
No details were provided regarding the preparation of diet.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily
Dose / conc.:
160 mg/kg bw/day (nominal)
Dose / conc.:
2 670 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10
Control animals:
not specified
Details on study design:
No details on study design are available in the source.

Positive control:
no
Observations and examinations performed and frequency:
No details are provided regarding it.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Other examinations:
No details are provided regarding it.

Clinical signs:
not specified
Mortality:
mortality observed, treatment-related
Description (incidence):
Death of animals was observed at 1.28 g/kg bw daily dose.
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Altered kidney or liver weight was observed at minimum daily dose of 0.64 g/kg bw.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Microscopic lesions were observed at 1.28 g/kg bw daily dose.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Details on results:
Death of animals was observed at 1.28 g/kg bw daily dose. No reduced growth and appetite were observed in animals, however, altered kidney or liver weight was found at minimum daily dose of 0.64 g/kg bw. Microscopic lesions and death were also observed at least daily dose of 1.28 g/kg bw.
Key result
Dose descriptor:
NOAEL
Effect level:
320 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: Based on mortality, altered liver or kidney weights and microscopic lesions at higher dose levels.
Conclusions:
In a range finding toxicity study, administration of monoethanolamine to rats in diet at dose levels of 160-2670 mg/kg bw/day or 90 days resulted in a NOAEL of 320 mg/kg bw/day (based on mortality, altered liver or kidney weights and microscopic lesions at higher dose levels). The LD50 value of test substance was 2.74 (95% CL: 2.39-3.15) g/kg bw.
Executive summary:

A range finding repeated dose toxicity test was performed with monoethanolamine (MEA) in rats. The test substance was administered via diet to groups of ten rats at doses in the range of 160-2670 mg/kg bw/ day for 90 days. No reduced growth and food consumption were observed in animals. Altered kidney and liver weight were observed at dose of 640 mg/kg bw/day. Microscopic lesions and mortality were also observed at 1280 mg/kg bw/day. Under the study conditions, the systemic toxicity NOAEL was established at 320 mg/kg bw/day (Smyth, 1951).

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Principles of method if other than guideline:
The purpose of this study is to determine the sub-acute toxicity of C10-13 sodium linear alkyl benzene sulfonate (LAS-Na) in CRJ-SD rats. LAS-Na was orally administered at 0 (distilled water), 125, 250 and 500 mg/kg bw to male and females rats for 29 and 30 days, respectively. General symptoms of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. After collection of blood samples, the animals were humanely euthanized, gross necropsy was performed and organs weights were determined.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: CRJ-SD rats
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc.
- Age at study initiation: 5 weeks
- Weight at study initiation: Not reported
- Fasting period before study: Not reported
- Housing: 3 male and 4 female animals per cage.
- Diet: Solid feed (CE-2, CLEA Japan, Inc.); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: After 1 week pre-housing, the animals were used in the experiment.

ENVIRONMENTAL CONDITIONS
- Temperature: 23 ± 2°C
- Humidity: 55 ± 5%
- Air changes: Not reported
- Photoperiod: Not reported

EXPERIMENTAL START DATE: Not reported
Route of administration:
oral: gavage
Details on route of administration:
The test substance was administered orally through a metallic gastric sonde.
Vehicle:
other: Distilled water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance was suspended in distilled water prior to dosing.

- DOSE VOLUME: 5 mL/kg bw

- Concentration in vehicle: 0, 250, 500, and 1000 mg/mL respectively for administration of doses of 0, 125, 250 and 500 mg/kg bw.
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
29 days for males and 30 days for females
Duration of treatment / exposure:
29 days for males and 30 days for females
Frequency of treatment:
Once daily
Dose / conc.:
125 mg/kg bw/day (actual dose received)
Remarks:
In distilled water (corresponding to 250 mg/mL)
Dose / conc.:
250 mg/kg bw/day (actual dose received)
Remarks:
In distilled water (corresponding to 500 mg/mL)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Remarks:
In distilled water (corresponding to 1000 mg/mL)
No. of animals per sex per dose:
Test group: 12 animals/sex/dose group
Control group: 15 male/group and 16 female/group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels for this study were selected based on the results of acute toxicity study conducted with LAS-Na in rats. 1/3 of the LD50 levels of LAS-Na, i.e. 500 mg/kg bw, was selected as the highest dose. The other two doses were set at common ratios of 2, whereby the medium and low doses were 250 (1/6 LD50) and 125 (1/12 LD50) mg/kg bw, respectively.
Positive control:
No
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Daily

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Time schedule for examinations: Twice/week

FOOD EFFICIENCY: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected was collected from the carotid artery using a cannula.
- Anesthetic used for blood collection: Yes, light ether anesthesia was used.
- Animals fasted: Not reported
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red and white blood cell count were measured using an auto-analyzer SMA-4A.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected from each animal and centrifuged (at 3000 rpm for 15 minutes) to obtain the serum samples for analysis.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: s-GOT, s-GPT, alkaline phosphatase, total protein, albumin, glucose, cholesterol, bilirubin, creatinine, urea nitrogen, inorganic phosphates and ions (Na+, K+, Ca2+, Mg2+ and Cl-)

URINALYSIS: Yes
- Time schedule: Not reported
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: pH, proteins, sugar, ketone bodies and occult blood

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

ORGAN WEIGHTS: The wet weight of liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary and the brain were measured. Based on this, the relative weights were calculated.

HISTOPATHOLOGY: No
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
At 500 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 125 and 250 mg/kg bw, however, the animals showed recovery at low and mid doses.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Male animals of all dose groups and female animals of high dose group (i.e. 500 mg/kg bw) exhibit body weight suppression.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Animals showed a dose dependent decline in the feed consumption.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Decreased feed efficacy was observed for male animals at 500 mg/kg bw.
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Effects at 125 mg/kg/bw: Decreased total proteins (only females), albumin (only females), Ca2+ levels (only males), and increased cholesterol (only males)
Effects at 250 mg/kg bw: Decreased s-GPT (both males and females), total proteins (only females), albumin (only females), Na+ (only males), K+ (only males), Ca2+ (both males and females), and Mg2+ (only males)
Effects at 500 mg/kg bw: Decreased s-GOT (only females), total proteins (only females), albumin (only females), glucose (only females), Ca2+ (both males and females), and increased alkaline phosphatase (only males) and urea nitrogen (only females).
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
At 500 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
CLINICAL SIGNS: At 500 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 125 and 250 mg/kg bw, however, the animals showed recovery at low and mid doses.

MORTALITY: No mortality was observed at any dose level.

BODY WEIGHT AND WEIGHT CHANGES: Body weight suppression was observed in animals (both males and females) belonging to the high dose group (i.e. 500 mg/kg bw).

FOOD CONSUMPTION AND COMPOUND INTAKE: Animals showed a dose dependent decline in the feed consumption. Decreased feed efficacy was observed for male animals at 500 mg/kg bw.

CLINICAL BIOCHEMISTRY: Few significant effects were observed in the biochemical parameters mainly at the mid and high dose level.

URINALYSIS: Results of urinalysis (pH, proteins, sugar, ketone bodies and occult blood) in the test animals were comparable to the control group.

ORGAN WEIGHT: The relative liver weight of female animals was significantly increased at 500 mg/kg bw (p<0.01). Apart from this, increase in relative weight of adrenal glands (p<0.05 for right side; p<0.01 for left side), testes (p<0.05) and brain (p<0.05) was also observed in male animals at 500 mg/kg bw. Decrease in the relative weight of liver (only females; at 125 mg/kg bw, p<0.05), right side kidney (only females; at 250 and 500 mg/kg bw, p<0.05), spleen (only males; at 500 mg/kg bw, p<0.05), heart (both males and females at 500 mg/kg bw, p<0.01; males at 250 mg/kg bw, p<0.05; females at 125 mg/kg bw, p<0.05) and thymus (only females; at 500 mg/kg bw, p<0.01) was also observed. Although, some significant effects were observed at the low and mid dose yet they were not dose-dependent.

GROSS PATHOLOGY: At 500 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group.
Key result
Dose descriptor:
NOAEL
Effect level:
125 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
organ weights and organ / body weight ratios
Remarks on result:
other: Based on reduced body weight and changes in organ weight (without histopathological correlation), clinical chemistry. The LOAEL was 250 mg/kg bw/day.

Table 1: Effect of LAS on average feed efficiency of rats after 1 month (po) exposure (Ito et al., 1978)

Sex Doses 2 weeks 4 weeks
Total intake (g) Net gain (g) Efficacy (%) Total intake (g) Net gain (g) Efficacy (%)
Males 0 346.5 108.8 31.4 747.6 199.1 25.6
125 322.7 97.5 30.2 692.3 183.2 25.4
250 316.4 92.8 29.4 690.2 177.5 24.6
500 284.2 76.5 27 633.5 152.4 22.7
Females 0 274.4 60.4 21.9 573.3 114.9 18.4
125 236.6 53.7 22.6 513.1 105.4 19.5
250 246.4 55.3 21.9 534.8 110.9 19.4
500 231.7 49.6 21.4 486.5 97.8 18

Table 2: Effect of LAS on hematological parameters of rats after 1 month (po) exposure (Ito et al., 1978)

Sex Doses RBC (106/mm3) Hb (g/dL) Ht (%) WBC (103/mm3)
Males 0 7.5 ± 0.11 16.5 ± 0.17 48.8 ± 0.63 16.5 ± 1.04
125 7.4 ± 0.08 16.0 ± 0.13 47.4 ± 0.49 17.4 ± 1.12
250 7.4 ± 0.17 15.9 ± 0.36 47.4 ± 1.13 17.8 ± 1.30
500 7.5 ± 0.07 16.0 ± 0.12 48.1 ± 0.34 18.8 ± 1.41
Females 0 6.6 ± 0.15 15.2 ± 0.18 44.0 ± 0.65 12.1 ± 0.49
125 6.6 ± 0.15 15.2 ± 0.15 44.4 ± 0.56 12.0 ± 0.87
250 6.6 ± 0.11 15.2 ± 0.13 44.5 ± 0.51 13.5 ± 0.91
500 6.8 ± 0.06 15.2 ± 0.10 44.2 ± 0.41 14.5 ± 1.31

Mean ± SE

* Significantly different from control (p<0.05)

** Significantly different from control (p<0.01)

Table 3a: Effect of LAS on biochemical parameters of rats after 1 month (po) exposure (Ito et al., 1978)

Sex Dose (mg/kg) sGOT sGPT ALP TP Alb Glu Chol
mu/mL mu/mL mu/mL mg% mg% mg% mg%
Males 0 137.0± 5.4 44.5 ± 1.6 294.8 ± 11.6 5.4 ± 0.08 3.3 ± 0.05 228.7 ± 5.8 55.4 ± 1.27
125 134.3 ± 8.4 39.5 ± 1.9 280.2 ± 17.5 5.1 ± 0.14 3.2 ± 0.08  228.9 ± 9.0 60.8* ± 2.38
250 133.3 ± 7.3 39.3 ± 1.5* 311.8 ± 26.6 5.1 ± 0.27 3.1 ± 0.16 225.3 ± 11.5 54.5 ± 2.48
500 145.4 ± 7.3 45.0 ± 2.1 374.0* ± 30.0 5.2 ± 0.07 3.3 ± 0.04 218.3 ± 3.5 53.4 ± 1.25
Females 0 130.3 ± 6.5 48.5 ± 1.1 184.9 ± 8.2  6.1 ± 0.08 3.8 ± 0.05 224.1 ± 5.7 67.7 ± 2.24
125 141.0 ± 7.3 44.4 ± 1.1  205.2 ± 10.0 5.8** ± 0.7 3.7* ± 0.05 231.7 ± 8.4 63.9 ± 1.89
250 134.5 ± 7.1 39.9 ± 0.8* 182.5 ± 11.5 5.7* ± 0.16 3.5** ± 0.09 216.2 ± 7.1 64.9 ± 2.81
500 162.5* ± 10.7 44.1 ± 2.0 222.7 ± 17.9 5.7** ± 0.08 3.6** ± 0.05 207.8* ± 5.5 62.3 ± 2.30

Mean ± SE

* Significantly different from control (p<0.05)

** Significantly different from control (p<0.01)

Table 3b: Effect of LAS on biochemical parameters of rats after 1 month (po) exposure (Ito et al., 1978)

Sex Dose (mg/kg) T-Bili Creat BUN InP Na K Ca Mg Cl
mg% mg% mg% mg% meg/L meg/L mg/dL mg/dL meg/L
Males 0 0.9 ± 0.03 0.5 ± 0.01 17.4 ± 0.99 6.8 ± 0.14 147.6 ± 1.7 5.0 ± 0.10 5.1 ± 0.07 2.5 ± 0.04 102.5 ± 0.62
125 0.8 ± 0.04 0.5 ± 0.02 17.1 ± 1.32 6.8 ± 0.16 142.3 ± 2.0 4.8 ± 0.14 4.6* ± 0.14 2.5 ± 0.10 103.7 ± 0.65
250 0.8 ± 0.04 0.4 ±0.03 17.0 ± 0.09 6.4 ± 0.26 135.8* ± 3.1 4.5** ± 0.11 4.6** ± 0.15 2.2** ± 0.05 104.6 ± 0.77
500 0.9 ± 0.02 0.5 ± 0.02 19.3 ± 1.17 6.8 ± 0.10 141.5 ± 2.9 5.0 ± 0.17 4.8* ± 0.12 2.4 ± 0.06 104.4 ± 1.22
Females 0 0.9 ± 0.09 0.5 ± 0.02 17.9 ± 0.82 6.2 ± 0.14 143 ± 1.3 4.5 ± 0.09 5.2 ± 0.08 2.5 ± 0.07 105.4 ± 0.43
125 0.8 ± 0.02 0.5 ± 0.01 17.9 ± 1.51 6.3 ± 0.23 143.4 ± 1.9 4.5 ± 0.08 5.0 ± 0.11 2.5 ± 0.07 106.8 ± 0.66
250 0.8 ± 0.05 0.5 ± 0.01 18.7 ± 0.99 6.3 ± 0.22 145.3 ± 1.5 4.4 ± 0.15 4.9* ± 0.06 2.5 ± 0.06 106.8 ± 0.54
500 0.9 ± 0.06 0.6 ± 0.02 24.0* ± 1.41 6.1 ± 0.21 142.6 ± 1.2 4.5 ± 0.08 4.9** ± 0.08 2.4 ± 50.0 106.6 ± 0.42

Mean ± SE

* Significantly different from control (p<0.05)

** Significantly different from control (p<0.01)

Table 4a: Effect of LAS on organ weight of rats after 1 month (po) exposure (Ito et al., 1978)

Sex Dose (mg/kg) Body weight (g) Liver Kidney Spleen Adrenal gland
Right Left Right Left
wt (g) % wt (g) % wt (g) % wt (g) % wt (mg) % wt (mg) %
Males 0 370.7± 6.0 14.60 ± 0.39 3.94 ± 0.08 1.38 ± 0.03 0.37 ± 0.01 1.38 ± 0.04 0.37 ± 0.01 0.70 ± 0.03 0.19 ± 0.01 25.6 ± 1.2 6.91 ± 0.32 25.8 ± 1.0 6.97 ± 0.26
125 355.1 ± 7.5 13.72±0.33 3.87 ± 0.06 1.36 ± 0.04 0.39 ± 0.01 1.33 ± 0.04 0.38 ± 0.01 0.68 ± 0.03 0.19 ± 0.01 24.4 ± 1.8 6.88 ± 0.48 25.3 ± 1.1 7.16 ± 0.37
250 349.8* ± 7.6 13.65 ± 0.42 3.90 ± 0.09 1.31 ± 0.03 0.38 ± 0.01 1.29 ± 0.03 0.37 ± 0.01 0.62 ± 0.03 0.18 ± 0.01 24.7 ± 1.4 7.06 ± 0.34 25.4 ± 1.5 7.24 ±0.35
500 324.4** ±9.0 13.37 ± 0.57 4.11 ± 0.11 1.21** ± 0.04 0.37 ± 0.01 1.20** ± 0.04 0.37 ± 0.01 0.53** ± 0.02 0.16* ± 0.01 26.2 ± 1.0 8.15* ± 0.40 28.5* ± 0.6 8.88** ± 0.36
Females 0 262.6± 8.0 10.84 ± 0.51 4.11 ± 0.08 1.09 ± 0.04 0.41 ± 0.01 1.08 ± 0.05 0.41 ± 0.2 0.55 ± 0.02 0.21 ± 0.01 34.0 ± 1.2 13.00 ± 0.38 35.8 ± 1.3 13.63 ± 0.30
125 258.0 ± 3.5 10.03 ± 0.22 3.89* ± 0.05 0.98* ± 0.03 0.38* ± 0.01 0.95* ± 0.02 0.37* ± 0.01 0.57 ± 0.02 0.22 ± 0.01 31.7 ± 1.1 12.27 ± 0.37 32.1 ± 1.5 12.43 ± 0.54
250 261.5 ± 4.5 10.56 ± 0.30 4.04 ± 0.08 1.00 ± 0.03 0.38* ± 0.01 0.96 ± 0.03 0.37* ± 0.01 0.56 ± 0.03 0.22 ± 0.01 31.0 ± 1.8 11.94 ± 0.84 33.4 ± 1.7 12.83 ± 0.76
500 249.7* ± 3.1 11.58 ± 0.21 4.64** ± 0.08 0.97** ± 0.02 0.39* ± 0.01 0.95* ± 0.02 0.38 ± 0.01 0.51 ± 0.01 0.21 ± 0.01 32.0 ± 1.9  12.79 ± 0.69 32.3 ± 2.0 12.89 ± 0.75

Mean ± SE

* Significantly different from control (p<0.05)

** Significantly different from control (p<0.01)

%: X10-3

Table 4b: Effect of LAS on organ weights of rats after 1 month (po) exposure (Ito et al., 1978)

Sex Dose (mg/kg) Lung Heart Thymus Testis/Ovarium Pituitary Brain
Right Left
wt (g) % wt (g) % wt (g) % wt (mg) % wt (mg) % wt (mg) % wt (g) %
Males 0 1.23± 0.02 0.33± 0.01 1.27 ± 0.05 0.34 ± 0.01 0.52 ± 0.02 0.14 ± 0.01 1.53 ± 0.03 0.41 ± 0.01 1.51 ± 0.03 0.41 ± 0.01 10.4 ± 0.4 2.81 ± 0.11 1.84 ± 0.03 0.50 ± 0.01
125 1.23 ± 0.03 0.35 ± 0.01 1.18 ± 0.03 0.33 ± 0.01 0.56 ± 0.05 0.16 ± 0.01 1.48 ± 0.03 0.42 ± 0.01 1.51 ± 0.03 0.43 ± 0.01 11.1 ± 1.1 3.14 ± 0.32 1.83 ± 0.03 0.52 ± 0.01
250 1.20 ± 0.03 0.35 ± 0.01 1.41 ± 0.03 0.33* ± 0.01 0.48 ± 0.02 0.14 ± 0.01 1.49 ± 0.04 0.43 ± 0.01 1.49 ± 0.03 0.43 ± 0.01 10.9 ± 0.7 3.10 ± 0.16 1.82 ± 0.02 0.52 ± 0.01
500 1.13* ± 0.03 0.35 ± 0.01 0.97** ± 0.03 0.30** ± 0.01 0.41** ± 0.03 0.13 ± 0.01 1.49 ± 0.04 0.46* ± 0.02 1.49 ± 0.04 0.46* ± 0.02 9.6 ± 0.4 2.99 ± 0.16 1.77* ± 0.02 0.55* ± 0.02
Females 0 1.07 ± 0.03 0.41 ± 0.01 0.96 ± 0.03 0.37 ± 0.01 0.59 ± 0.02 0.23 ± 0.01 44.8 ± 2.9 17.04 ± 0.95 47.8 ± 3.5 18.13 ± 1.07 11.8 ± 0.4 4.53 ± 0.18 1.80 ± 0.02 0.69 ± 0.02
125 1.10 ± 0.05 0.43 ± 0.02 0.88** ± 0.01 0.34* ± 0.01 0.54 ± 0.02 0.21 ± 0.01 46.2 ± 2.5 17.94 ± 1.04 43.7 ± 1.8 16.93 ± 0.64 12.3 ± 0.8 4.79 ± 0.30 1.78 ± 0.02 0.69 ± 0.01
250 1.11 ± 0.03 0.42 ± 0.01 0.92 ± 0.02 0.35 ± 0.01 0.55 ± 0.03 0.21 ± 0.01 46.5 ± 2.8 17.73 ± 0.94  47.6 ± 2.8 18.12 ± 0.91 11.8 ± 0.8 4.53 ± 0.30 1.76 ± 0.02 0.68 ± 0.01
500 1.01 ± 0.02 0.41 ± 0.01 0.79** ± 0.03 0.32** ±0.01 0.44** ± 0.04 0.18** ± 0.02 46.3 ± 3.5 18.51 ± 1.29 46.2 ± 2.7 18.44 ± 0.97 11.8 ± 0.9 4.71 ± 0.34 1.75* ± 0.01 0.70 ± 0.01

Mean ± SE

* Significantly different from control (p<0.05)

** Significantly different from control (p<0.01)

%: X10-3

Conclusions:
Oral administration of C10-13 sodium linear alkylbenzene sulphonate (LAS-Na) to CRJ-SD rats at 0, 125, 250 and 500 mg/kg bw for 1 month resulted in a NOAEL of 250 mg/kg bw, based on reduced body weight, changes in organ weight (without histopathological correlation) and clinical chemistry at the mid and high doses. The LOAEL was 500 mg/kg bw.
Executive summary:

A short-term repeated dose toxicity study was conducted with C10-13 LAS, sodium salt in CRJ-SD rats. The test substance was administered via oral gavage to groups of male and female rats for 28 days at dose levels of 0, 125, 250 and 500 mg/kg/day. Clinical signs of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters, following which the animals were humanely euthanized, gross necropsy was performed, the organs weights were determined and histopathological examination of the major organs were performed. Body weight gain was suppressed, some serum biochemical measures were different from the controls, and some organ weights were either decreased (spleen, heart, thymus) or increased (liver) at the high dose. No mortalities or histopathological abnormalities were observed. No adverse effects were up to 125 mg/kg bw/day mg/kg/day. Under the study conditions, the systemic toxicity NOAEL was determined to be 125 mg/kg/day based on reduced body weight, changes in organ weight (without histopathological correlation) and clinical chemistry at the mid and high doses (Ito, 1978a). 

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Principles of method if other than guideline:
The purpose of this study is to determine the sub chronic toxicity of LAS in Wistar JCL rats, focusing on the liver and kidneys. Male and female rats were maintained on either test diets (0, 0.6 and 1.8%) or drinking water (0, 0.07 and 0.2%) for 9 months. 0.6 and 1.8% rats in drinking water study exhibited severe weight loss so LAS administration was stopped after 2 weeks. Mortality, clinical observations and body weights were recorded during the study. All the surviving animals were humanely euthanized at end of 9 months and gross necropsy, hematological, serum biochemical tests, enzyme tests on the liver and kidneys were performed and organs weights were measured. No histopathology was performed.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: Wistar JCL
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female rats were obtained from Shizuoka Agricultural Cooperative Association for
Laboratory Animals
- Age at study initiation: 4-weeks old
- Weight at study initiation: 100 - 124 g (male rats), 82 - 100 g (female rats)
- Fasting period before study: No
- Housing: 5 animals were housed per cage.
- Diet: CE-2 food (from CLEA Japan); ad libitum
- Water: ad libitum
- Acclimation period: 1 week before the start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 50 - 60%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: Not reported
Route of administration:
other: oral: drinking water and feed
Details on route of administration:
LAS was administered to animals by mixing 0.6 and 1.8% in CE-2 food (CLEA Japan) and dissolving to 0.07 and 0.2% in drinking water.
Vehicle:
other: Test substance was administered either in diet or drinking water
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
9 months
Frequency of treatment:
Continuous in diet or drinking water (ad libitum)
Dose / conc.:
0.6 other: %
Remarks:
in diet (corresponding to 300 mg/kg bw/day)
Dose / conc.:
1.8 other: %
Remarks:
in diet (corresponding to 900 mg/kg bw/day)
Dose / conc.:
0.07 other: %
Remarks:
in drinking water (corresponding to 85 mg/kg bw/day)
Dose / conc.:
0.2 other: %
Remarks:
in drinking water (corresponding to 145 mg/kg bw/day)
No. of animals per sex per dose:
Feeding study (mixed in diet): 8 animals/sex/dose
Drinking water study: 9 animals/sex/dose
Control animals:
yes, concurrent vehicle
yes, plain diet
Positive control:
No
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Compound intake: Yes
- Time schedule for examinations: Weekly

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: Weekly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 9 months, blood was collected from each animal after euthanizing.
- Anesthetic used for blood collection: No
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters: White blood cells (WBC), red blood cells (RBC), Hemoglobin (Hgb), Hematocrit (Hct), mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 9 months, blood was collected from each animal after euthanizing.
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters: Glutamate oxaloacetate transaminase (GOT), glutamate pyruvic transaminase (GPT), glucose content, urea nitrogen, total cholesterol, albumin, alkaline phosphatase (ALP) and cholinesterase

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER:
- Liver enzyme tests: Glucose 6-phosphatase (G6Pase), lactase dehydrogenase (LDH) and G6P-DH activity
- Kidney enzyme tests: G6Pase, LDH, GPT, GOT, ALP, acid phosphatase (ACP), Na, K-ATPase, and malate dehydrogenase (MDH)
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (However, no details in study report in mentioned)
Gross findings were obtained after euthanizing animals and organs were removed for organ weight measurements.

ORGAN WEIGHTS: Brain, heart, lungs, liver, spleen, kidneys, adrenal gland, testes, ovary, uterus, and appendix
HISTOPATHOLOGY: No
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Both female and male rats (drinking water study) exhibited a slight redness at the tips of their facial fur, and coarse fur all over their bodies.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was significant decrease in body weight gain in males and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
A significant reduction in WBC was observed in 0.6% (diet) male rats and in MCV and MCH was observed in 1.8% (diet) female rats compared to controls.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
There were significant alteration in cholesterol [decrease; all doses, except female rats of 0.07% dose group (drinking water)], GPT [0.6% dose group (diet) females)], GOT [1.8% dose group (diet) males], albumin [1.8% (diet) males] , ALP levels [male and female rats fed with 1.8% LAS-diet] and cholinesterase levels [in male rats fed with 1.8% LAS-diet].
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In males of 1.8% (diet), the absolute liver and testes weight was significantly decreased and the relative testes and liver weight was significantly increased and both absolute and relative spleen weight were also significantly reduced.
In females of 1.8% (diet), absolute and relative liver and caecum weight was significantly increased.
There was also significant decrease in absolute and relative heart weight in females of 0.6% (diet) and 0.07% (drinking water) dose groups.
Gross pathological findings:
not specified
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Liver enzyme tests:
Dietary study: G6Pase activity was reduced in 1.8% dose group males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group males and females, LDH activity was reduced in 0.6, 1.8% dose group. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group and GOT activity was reduced in 1.8% dose group animals.
Drinking water study: LDH activity was reduced in 0.2% dose group males. GOT and GPT activities were clearly reduced in males and GOT activity was increased in 0.07 and 0.2% dose group animals.

Renal enzyme tests:
Dietary study: A significant difference was also observed in G6Pase activity. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals
Drinking water study: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group. A significant difference was also observed in G6Pase activity.
Details on results:
CLINICAL SIGNS: Both female and male rats consuming LAS-containing water exhibited a slight redness at the tips of their facial fur, and coarse fur all over their bodies.

BODY WEIGHT AND WEIGHT CHANGES: There was significant decrease in body weight gain in male and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.

WATER CONSUMPTION AND COMPOUND INTAKE: Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.

HAEMATOLOGICAL FINDINGS: A significant reduction in WBC was observed in 0.6% (diet) male rats compared to controls, and a significant reduction in MCV and MCH was observed in 1.8% (diet) female rats compared to controls.

CLINICAL BIOCHEMISTRY: Except female rats of 0.07% dose group (drinking water), a significant reduction or a reduction in cholesterol was observed in male and female rats of all dose groups compared to controls. GPT was significantly reduced in 0.6% dose group (diet) females, and reduced in females of other treatment groups. GOT was significantly reduced in 1.8% dose group (diet) males, and reduced in both females and males of other treatment groups. Albumin was significantly reduced in 1.8% (diet) males and reduced in males of 0.2% dose group (drinking water) and females of 1.8% dose group (diet). There was significant increase in ALP levels in male and female rats fed with 1.8% LAS-diet and cholinesterase levels in male rats fed with 1.8% LAS-diet.

ORGAN WEIGHT: In males of 1.8% (diet), the absolute liver and testes weight was significantly decreased and the relative testes and liver weight was significantly increased and both absolute and relative spleen weight were also significantly reduced. In females of 1.8% (diet), absolute and relative liver and caecum weight was significantly increased. There was also significant decrease in absolute and relative heart weight in females of 0.6% (diet) and 0.07% (drinking water) dose groups.

LIVER ENZYME TESTS: G6Pase activity was reduced in 1.8% dose group (diet) males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group (diet) males and females, where the percentage reduction was greater in 1.8% dose group (diet) animals. LDH activity was clearly reduced in 0.6, 1.8% dose group (diet), and 0.2% dose group (drinking water) males, but reduced in only 1.8% dose group (diet) females. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group (diet) animals, and GOT activity was increased in 0.07 and 0.2% dose group (drinking water) animals but reduced in 1.8% dose group (diet) animals.

RENAL ENZYME TESTS: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group (drinking water) animals, and also reduced in other male treatment groups. A significant difference was also observed in G6Pase activity, where the reduction observed was associated with an increase in amount consumed. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals, and G6Pase and LDH activity were also reduced in other treatment groups.
Key result
Dose descriptor:
LOAEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
act. ingr.
Remarks:
(dietery study)
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
water consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
other: Liver and kidney enzyme levels
Remarks on result:
other: Adverse effects were observed at all dose levels
Key result
Dose descriptor:
NOAEL
Effect level:
85 mg/kg bw/day (nominal)
Based on:
act. ingr.
Remarks:
(drinking water study)
Sex:
male/female
Basis for effect level:
clinical biochemistry
other: Liver and kidney enzyme levels
Remarks on result:
other: Based on significant decreases in the activities of glutamate-oxalate transaminase and lactate dehydrogenase in males and significant decrease in renal Na,K-ATPase in males and females at 145 mg/kg bw/day (0.2% drinking water).
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
145 mg/kg bw/day (nominal)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
145 mg/kg bw/day (nominal)
System:
hepatobiliary
Organ:
liver
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Body weight gain, food, water and sample consumption of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  No. of rats  Initial body weight (g) Final Body weight (g) Body weight gain (%) Food (g/rat/day) Water (g/rat/day) Sample (g/kg BW/day)
Male Control 8 113.1± 1.156 421.6 ± 8.542 372.6 ± 5.889 15 23 -
0.6 F 8 110.6 ± 0.925 413.5 ± 7.356 373.8 ± 6.380 16 23 0.234
1.8 F 8 109.2 ± 2.218 341.1 ± 8.008  312.5 ± 6.518** 14 33 0.747
0.07 W  9 110.6 ± 1.572 411.0 ± 5.255 371.9 ± 6.908  17 29 0.051
0.2 W 9 108.3 ± 1.900 385.1 ± 5.453 356.1 ± 6.346 ** 15 33 0.148
Female Control 8 92.3 ± 1.592 212.6 ± 4.508 230.5 ± 5.420 10 18 -
0.6 F 8 92.1 ± 1.663 216.1 ± 7.705 234.3 ± 5.753 10 17 0.287
1.8 F 8 90.6 ± 1.463 184.8 ± 4.278 204.3 ± 5.791 ** 9 18 0.969
0.07 W  9 92.7 ± 1.770 206.0 ± 3.555 222.4 ± 4.478 11 20 0.082
0.2 W 9 91.6 ± 1.893 209.3 ± 5.383 228.4 ± 3.857 12 18 0.173

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 2: Blood components of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  WBC ( x 103) RBC ( x 106) HGB (g) HCT (%) MCV (µ3) MCH (µµg) MCHC (%)
Male Control 5.5±0.414 5.7 ± 0.142 10.9 ± 0.195 28.0 ± 0.663 49.0 ± 0.189 19.5 ± 0.247 39.5 ± 0.504
0.6 F 4.2 ± 0.221 * 5.9 ± 0.159 11.2 ± 0.291 29.2 ± 1.059 49.6 ± 0.778 19.4 ± 0.436 38.9 ± 0.851
1.8 F 4.9 ± 0.461 5.8 ± 0.076 10.7 ± 0.118 27.8 ± 0.561 49.2 ± 0.250  19.1 ± 0.268 38.5 ± 0.499
0.07 W  4.7 ± 0.209 5.9 ± 0.072 11.3 ± 0.094 29.1 ± 0.317 49.2 ± 0.147 19.1 ± 0.210  38.6 ± 0.447
0.2 W 4.9 ± 0.304 6.0 ± 0.119 11.3 ± 0.141 29.3 ± 0.566 48.8 ± 0.200 19.1 ± 0.238 38.7 ± 0.493
Female Control 2.6 ± 0.297 5.6 ± 0.163 11.0 ± 0.243 28.8 ± 0.859 51.7 ± 0.164 20.0 ± 0.364  38.4 ± 0.689
0.6 F 2.5 ±0.150 5.7 ± 0.265 11.2 ± 0.485 29.3 ± 1.400 51.2 ± 0.313 19.8 ± 0.356 38.5 ± 0.699
1.8 F 3.7 ± 0.556 5.6 ± 0.294 10.5 ± 0.599 28.2 ± 1.546 50.0 ± 0.378 ** 18.9 ± 0.193* 37.5 ± 0.294
0.07 W  2.4 ± 0.213 6.0 ± 0.302 11.5 ± 0.556 30.5 ± 1.551 51.5 ± 0.176  19.5 ± 0.159 37.7 ± 0.299
0.2 W 2.7 ± 0.328 5.6 ± 0.100 11.0 ± 0.176 28.9 ± 0.624 52.0 ± 0.463 20.0 ± 0.349 38.3 ± 0.601

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 3: Biochemical values in serum of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  GOT (K.U/mL) GPT (K.U/mL) GLU (mg/dL) UN (mg/dL) CHO (mg/dL) ALB (g/dL) ALP ((KA-U/dL) ChE (ΔpH)
Male Control 151.3 ± 7.285   107.5 ± 8.177 163.0 ± 11.488 21.0 ± 0.333 77.5 ± 3.417  4.87 ± 0.055 41.3 ± 1.308 0.166 ± 0.010
0.6 F 133.2 ± 5.573 102.3 ± 8.485 132.2 ± 4.139* 21.0 ± 0.684 71.6 ± 2.314 47.2 ± 0.075 44.7 ± 2.782  0.187 ± 0.013
1.8 F 126.7 ± 6.439* 93.0 ± 3.665 144.7 ± 10.099 23.1 ± 0.490** 53.0 ± 1.282** 44.7 ± 0.068** 82.3 ± 3.731** 0.281 ± 0.007 **
0.07 W  142.8 ± 10.111 108.5 ± 12.381 131.6 ± 3.088* 22.8 ± 0.647* 68.2 ± 2.527* 47.5 ± 0.059 44.0 ± 1.239 0.131 ± 0.015
0.2 W 143.8 ± 9.041 103.3 ± 9.513 165.1 ± 14.043 21.0 ± 1.462 62.5 ± 2.534** 43.2 ± 0.251 34.4 ± 3.193 0.171 ± 0.008
Female Control 142.3 ± 14.359 104.2 ± 12.474 135.5 ± 5.246 23.2 ± 0.793 95.8 ± 4.778 47.1 ± 0.265 45.0 ± 5.728 0.963 ± 0.073
0.6 F 116.1 ± 5.244 68.7 ± 2.670* 126.4 ± 3.657 21.5 ± 0.602 86.7 ± 4.230 4.85 ± 0.095 40.7 ± 1.539 1.056 ± 0.023
1.8 F 133.6 ± 4.007 88.1 ± 4.362 130.8 ± 3.945 26.7 ± 1.935 67.8 ± 7.472** 42.8 ± 0.097 73.8 ± 11.808* 0.861 ± 0.053
0.07 W  121.2 ± 7.003 74.2 ± 13.529 127.2 ± 2.035 21.9 ± 0.872 95.0 ± 3.755 49.0 ± 0.187 45.6 ± 12.960 0.881 ± 0.171
0.2 W 128.5 ± 12.209 85.7 ± 9.362 136.1 ± 10.070 21.7 ± 0.950 77.2 ± 8.826 46.8 ± 0.155 44.0 ± 5.756 1.053 ± 0.042

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 4:  Organ weight of male and female rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  Brain  Heart Lung Liver Spleen Kidney (R) Kidney (L) Adrenal (R) Adrenal (L) Testis(R) Testis(L) Ovary (R) Ovary (L) Caecum Uterus
g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g
Male Control 1.987 ± 0.011 0.463 ± 0.008 1.026 ± 0.018 0.239 ± 0.005 1.263 ± 0.033 0.294 ± 0.008 12.7 ± 0.341 2.9 ± 0.039 0.698 ± 0.014 0.162 ± 0.003 1.162 ± 0.40 0.270 ± 0.008 1.184 ± 0.041 0.275 ± 0.007 18.3 ± 1.117 4.2 ± 0.217 18.1 ± 0.934  4.2 ± 0.225 1.591 ± 0.023 0.370 ± 0.006 1.643 ± 0.027 0.382 ± 0.005 - - - - 1.238 ± 0.068 0.281 ± 0.015 - -
0.6 F 1.982 ± 0.011 0.473 ± 0.007 0.971 ± 0.022* 0.231 ± 0.004 1.269 ± 0.042 0.302 ± 0.005 12.9 ± 0.310 3.0 ± 0.049 0.682 ± 0.019 0.162 ± 0.002 1.210 ± 0.026 0.290 ± 0.006 1.214 ± 0.032 0.289 ± 0.007 19.6 ± 1.900 4.7 ± 0.435 17.6 ± 1.602 4.2 ± 0.416 1.563 ± 0.019 0.373 ± 0.004 1.585 ± 0.027 0.378 ± 0.007 - - - - 1.303 ± 0.057 0.305 ± 0.010 - -
1.8 F 1.827 ± 0.131 0.521 ± 0.037 0.825 ± 0.026* 0.235 ± 0.003 1.040 ± 0.025** 0.295 ± 0.002 11.5 ± 0.352* 3.2 ± 0.043** 0.501 ± 0.163** 0.143 ± 0.003** 0.977 ± 0.022** 0.279 ± 0.007 0.982 ± 0.017** 0.281 ± 0.007 17.5 ± 1.370 5.0 ± 0.481 17.0 ± 0.654 4.8 ± 0.146 1.455 ± 0.027** 0.415 ± 0.004** 1.491 ± 0.033** 0.425 ± 0.004** - - - - 2.205 ± 0.141 ** 0.647 ± 0.032** - -
0.07 W  1.995 ± 0.016 0.471 ± 0.006 0.981 ± 0.031 0.232 ± 0.008 1.287 ± 0.021 0.304 ± 0.007 13.2 ± 0.258 3.0 ± 0.052 0.682 ± 0.009 0.161 ± 0.002 1.221 ± 0.018 0.288 ± 0.005 1.254 ± 0.012 0.296 ± 0.005* 18.6 ± 0.680  4.4 ± 0.156 19.2 ± 1.024 4.5 ± 0.262 1.606 ± 0.022 0.379 ± 0.006 1.566 ± 0.056 0.370 ± 0.015 - - - - 1.338 ± 0.040 0.314 ± 0.009 - -
0.2 W 1.881 ± 0.108 0.478 ± 0.028 0.975 ± 0.183 0.247 ± 0.003 1.154 ± 0.037* 0.293 ± 0.007 11.6 ± 0.184 2.9 ± 0.042 0.595 ± 0.013** 0.150 ± 0.002 1.124 ± 0.025 0.286 ± 0.007 1.154 ± 0.031 0.293 ± 0.008 18.4 ± 0.914  4.6 ± 0.232 17.5 ± 1.309 4.4 ± 0.354 1.483 ± 0.059 0.377 ± 0.016 1.589 ± 0.013 0.404 ± 0.005 - - - - 1.210 ± 0.035 0.309 ± 0.008 - -
Female Control 1.853 ± 0.023 0.846 ± 0.016 0.647 ± 0.013 0.294 ± 0.003 0.870 ± 0.026 0.397 ± 0.012 6.2 ± 0.150 2.8 ± 0.051 0.441 ± 0.016 0.201 ± 0.006 0.650 ± 0.021 0.296 ± 0.006 0.650 ± 0.021 0.296 ± 0.008 24.8 ± 1.381  11.2 ± 0.443 25.7 ± 1.385 11.7 ± 0.487 - - - - 29.3 ± 2.738 13.4 ± 1.226 31.2 ± 4.007 14.2 ± 1.857 0.828 ± 0.010 0.377 ± 0.005 0.687 ± 0.037 0.313 ± 0.017
0.6 F 1.859 ± 0.011 0.845 ± 0.027 0.595 ± 0.019* 0.269 ± 0.006** 0.810 ± 0.032 0.367 ± 0.015 6.6 ± 0.355 2.9 ± 0.061 0.425 ± 0.010 0.193 ± 0.009 0.648 ± 0.023 0.292 ± 0.004 0.655 ± 0.026 0.296 ± 0.008 25.8 ± 1.563 11.7 ± 0.820 24.8 ± 1.641 11.3 ± 0.863 - - - - 23.3 ± 3.504 10.8 ± 1.740 32.8 ± 3.120 14.9 ± 1.380 0.859 ± 0.064  0.381 ± 0.015 0.660 ± 0.033 0.301 ± 0.020
1.8 F 1.788 ± 0.031 0.954 ± 0.015** 0.455 ± 0.008 0.242 ± 0.002 0.761 ± 0.028 0.405 ± 0.009 7.1 ± 0.195** 3.8 ± 0.082** 0.351 ± 0.012 ** 0.187 ± 0.004 0.580 ± 0.015 0.309 ± 0.003 0.586 ± 0.021* 0.312 ± 0.007 19.6 ± 1.981* 10.3 ± 0.924 21.3 ± 1.487* 11.3 ± 0.609 - - - - 25.3 ± 2.853 13.4 ± 1.417 31.5 ± 2.322 16.7 ± 1.165 1.436 ± 0.080** 0.775 ± 0.039** 0.588 ± 0.066 0.312 ± 0.034
0.07 W  1.854 ± 0.019 0.844 ± 0.016 0.611 ± 0.011* 0.277 ± 0.004** 0.801 ± 0.027 0.365 ± 0.014 6.3 ± 0.139 2.8 ± 0.063 0.424 ± 0.007 0.193 ± 0.004 0.669 ± 0.017 0.304 ± 0.006 0.677 ± 0.010 0.308 ± 0.005 22.0 ± 1.619 10.0 ± 0.720 21.0 ± 1.763 9.8 ± 0.790 - - - - 30.0 ± 1.490 13.6 ± 0.780 31.7 ± 2.300 12.2 ± 1.200 0.785 ± 0.051 0.358 ± 0.024 0.729 ± 0.051 0.331 ± 0.022
0.2 W 1.691 ± 0.112 0.810 ± 0.053 0.593 ± 0.015 0.287 ± 0.004 0.817 ± 0.040 0.393 ± 0.021 6.1 ± 0.220 2.9 ± 0.071 0.428 ± 0.014 0.205 ± 0.006 0.651 ± 0.015 0.312 ± 0.006 0.670 ± 0.020 0.321 ± 0.008* 23.2 ± 1.495 11.1 ± 0.677 24.3 ± 1.154 11.6 ± 0.431 - - - - 28.4 ± 1.633 13.6 ± 0.821 28.6 ± 1.900 13.7 ± 0.830 0.800 ± 0.045 0.379 ± 0.026 0.644 ± 0.032 0.308 ± 0.014

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 5: Liver enzyme activities of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  G6 Pase (µmoles Pi/mg protein /hr G6P-DH (x 10-3)ΔE/mg protein / min LDH ΔE/mg protein / min GPT (x 102) KU/mg protein  GOT (x 102) KU/mg protein 

Male

Control 3.77 ± 0.22 60.3 ± 9.9  11.33 ± 0.53 4.06 ± 0.345 5.80 ± 0.525
0.6 F 3.82 ± 0.50 40.9 ± 9.1 ** 10.28 ± 0.84* 3.94 ± 0.484 5.49 ± 0.810
1.8 F 2.60 ± 0.015** 36.0 ± 7.7** 9.26 ± 0.99** 3.09 ± 0.225** 4.95 ± 0.510**
0.07 W  3.57 ± 0.36 68.0 ± 15.1 10.92 ± 0.91 3.89 ± 0.360 5.61 ± 0.410
0.2 W 3.71 ± 0.35 56.8 ± 14.9 9.28 ± 0.71** 4.10 ± 0.420 6.16 ± 0.545
Female Control 2.73 ± 0.57  143.6 ± 30.0 70.6 ± 11.6 2.71 ± 0.405 4.91 ± 0.635
0.6 F 2.67 ± 0.51 100.7 ± 39.9* 63.4 ± 14.2 2.60 ± 0.505 4.30 ± 0.560
1.8 F 2.17 ± 0.15* 58.5 ± 12.2* 62.2 ± 4.7 2.37 ± 0.225 4.05 ± 0.450 **
0.07 W  2.61 ± 0.20 138.3 ± 41.0 65.8 ± 8.5 2.57 ± 2.290 5.61 ± 0.410*
0.2 W 2.76 ± 0.28 120.9 ± 27.6 66.5 ± 11.3 2.89 ± 0.195

6.16 ± 0.545**

Values: mean ± S.D.

*p<0.05

**p<0.01

Table 6: Kidney enzyme activities of rats on administration of LAS for 9 months  (Yoneyama et. al., 1976)

Sex  Groups  ALP (µmoles/mg/hr) ACP (µmoles/mg/hr) G6Pase (µmoles/mg/hr) Na, K-A Tpase (µmoles/mg/hr) GPT (KU/mL) GOT (KU/mL) LDH (ΔE/mg/min) MDH (ΔE/mg/min)
Male Control 16.37 ± 1.984 1.80 ± 0.188 3.97 ± 0.590 3.94 ± 0.613 23.99 ± 5.300 192.11 ± 35.741 0.339 ± 0.043 2.308 ± 0.0447
0.6 F 14.44 ± 2.046 1.72 ± 0.141 3.92 ± 0.618 3.63 ± 0.640 23.16 ± 5.885 199.95 ± 22.869 0.334 ± 0.050 2.325 ± 0.575
1.8 F 15.74 ± 1.933 1.78 ± 0.179 3.47 ± 0.732 3.32 ± 1.311 21.11 ± 4.690 194.18 ± 13.668 0.358 ± 0.070 2.354 ± 0.554
0.07 W  14.73 ± 2.387 1.85 ± 0.194 4.26 ± 0.547 3.52 ± 1.170  23.85 ± 4.640 207.39 ± 21.308 0.347 ± 0.098 2.617 ± 0.628
0.2 W 15.03 ± 1.169 1.66 ± 0.109 4.03 ± 0.428 3.09 ± 0.224* 24.44 ± 3.535 202.94 ± 15.708 0.362 ± 0.058 2.679 ± 0.740 
Female Control 15.94 ± 3.369 2.02 ± 0.182 3.96 ± 0.545 4.29 ± 1.027 28.27 ± 4.407 188.33 ± 21.189 0.350 ± 0.035 2.387 ± 0.601
0.6 F 17.11 ± 2.919 2.03 ± 0.179 3.63 ± 0.521 4.29 ± 0.656 30.26 ± 5.736 196.48 ± 21.273 0.338 ± 0.044 2.231 ± 0.433
1.8 F 16.86 ± 2.369 1.85 ± 0.195 3.13 ± 0.426** 3.19 ± 0.613* 32.60 ± 3.737 184.89 ± 11.276 0.291 ± 0.025** 2.181 ± 0.404
0.07 W  16.24 ± 2.381 1.98 ± 0.218 3.72 ± 0.443 4.14 ± 1.452 27.19 ± 3.319 198.09 ± 29.803 0.322 ± 0.035 2.052 ± 0.306
0.2 W 15.02 ± 3.244 1.90 ± 0.136 3.45 ± 0.651 4.59 ± 1.154 27.03 ± 4.430 187.36 ± 20.235 0.316 ± 0.052 2.070 ± 0.362 

Values: mean ± S.D.

*p<0.05

**p<0.01

Conclusions:
Administration of LAS to Wistar JCL rats by test diets at dose levels of 0, 0.6 and 1.8% for 9 months (focusing on the liver and kidneys) revealed an LOAEL of 0.6% (300 mg/kg bw/day in diet), based on adverse effects at all dose levels.
Administration of LAS to Wistar JCL rats in drinking water at dose levels of 0, 0.07 and 0.2% for 9 months (focusing on the liver and kidneys) revealed an NOAEL of 0.07% (85 mg/kg bw/day in drinking water), based on adverse effects at all dose levels, based on significant decreases in the activities of glutamate-oxalate transaminase and lactate dehydrogenase in males and significant decrease in renal Na,K-ATPase in males and females at 145 mg/kg bw/day (0.2% drinking water).
Executive summary:

A sub-chronic (9 months) repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in male and female Wistar JCL rats. The test substance was administered at the following dose levels: through diet at 0, 0.6 and 1.8% (equivalent to 0, 300 and 900 mg/kg bw/day) to 8 animals/sex/dose and via drinking water at 0, 0.07 and 0.2% (equivalent to 0, 85 and 145 mg/kg bw/day to 9 animals/sex/dose. Rats at the 0.6 and 1.8% dose group exhibited severe weight loss so LAS administration via diet was stopped after 2 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. Terminal blood collection was done for estimation of haematological and clinical chemistry parameters. Gross findings were observed after euthanizing animals and organs were removed for organ weight measurements. No histopathology was performed. Body weight gain was reduced at the highest dose group only. Significant decreases in transaminase activity and renal Na,K-ATPase were seen at 145 mg/kg bw/day. No significant haematological or organ weight changes were noted. Based on significant decrease in the liver and renal enzyme levels at 145 mg/kg bw/day, the systemic toxicity NOAEL was established at 85 mg/kg bw/day (Yoneyama, 1976).  

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1964
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Reason / purpose:
reference to same study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
(two dose levels are used in the study against the guideline which recommends at least three dose levels should be used.)
GLP compliance:
no
Remarks:
(pre-GLP)
Limit test:
no
Species:
rat
Strain:
other: FDRL strain (Wistar-derived)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Not specified
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: All animals were at its weaning stage
- Weight at study initiation: Not specified
- Fasting period before study: Not specified
- Housing: Animals were housed individually in raised-bottom, wire-mesh cages
- Diet: Purina laboratory chow, ralston purina co., St. Louis, Missouri; ad libitum
- Water: ad libitum
- Acclimation period: Not specified

No information was provided for food and water quality and environmental conditions.
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
- DIET PREPARATION
No details were provided regarding the preparation of diet. Since rats consume approximately 2.4 times as much diet in proportion to body weight when they are first weaned as when they are adult, lower concentrations were incorporated in their diets initially, and increments in concentration made at biweekly intervals so that the intake of the test materials was relatively constant throughout. By the 11th week on test, when the animals had attained maturity, the levels in the diet were equivalent to 1000 and 5000 ppm of LAS.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
12 weeks
Frequency of treatment:
daily
Dose / conc.:
0.05 other: g/kg bw/day
Dose / conc.:
0.25 other: g/kg bw/day
No. of animals per sex per dose:
15
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Not specified
- Rationale for animal assignment (if not random): Not specified
- Rationale for selecting satellite groups: No satellite group was included in the study.
- Post-exposure recovery period in satellite groups: not specified
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly
- Body weights were recorded for approximately half the rats of each sex.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Food consumption was recorded for approximately half the rats of each sex.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: Not examined

OPHTHALMOSCOPIC EXAMINATION: Not examined

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During 6th and 12th week of test
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: Half animals from each group (not subjected to body weights and food consumption analysis)
- Parameters: Hemoglobin, hematocrit, total and differential leukocyte counts were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: During 6th and 12th week on test
- Animals fasted: Not specified
- How many animals: Half animals from each group (not subjected to body weights and food consumption analysis)
- Parameters: Blood glucose and urea nitrogen were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: During 6th and 12th week on test
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Half animals from each group (not subjected to body weights and food consumption analysis)
- Parameters: Albumin, pH, glucose, and microscopic examination of the sediment were examined.

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
Following the 12-week clinical examinations, all surviving rats were euthanized by over dosage with sodium pentobarbital, and examined at necropsy for evidence of gross pathologic changes. The liver, kidneys, spleen, heart, adrenals, pituitary, and cecum of each rat were weighed. Representative specimens of all organs and tissues from each animal were preserved in formalin.

Microscopic examinations were made of hematoxylin-eosin stained sections of the liver, spleen, stomach, small intestine, large intestine, pancreas, kidney, bladder, adrenal, gonads, thyroid, pituitary, thymus, salivary gland, lymph node, heart, lung, femoral marrow, aorta, muscle, spinal cord, and brain of 5 rats of each sex in the control and 250 mg/kg bw/day groups. Microscopic examinations were also made of the liver, spleen, stomach, kidneys, and cecum of all animals in the 50 mg/kg bw/day groups.
Clinical signs:
no effects observed
Mortality:
not specified
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
Occasional signs of wetting over the posterior ventral region of were noted during the first 14 days for females fed with 0.25 g/kg bw/day.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The t values for relative liver weight in the females in the 0.25 g/kg bw/day LAS group indicated a statistical significant increase in weight of this organ, compared to the controls. The weights of other organs did not show significant differences from the controls.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The frequent finding at gross necropsy was hemorrhagic areas in the lungs, characteristic of the common type of pneumonitis seen in rats. The scattered incidence of fluid-containing kidneys of rats in some groups was also noticed. However, both of these findings were not associated with the test materials or dosage levels and was unrelated to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
50 other: mg/kg bw/day
Based on:
act. ingr.
Sex:
female
Basis for effect level:
organ weights and organ / body weight ratios
Remarks on result:
other: There was significant increase in relative liver weights in females fed with 250 mg/kg bw/day
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
250 other: mg/kg bw/day
System:
gastrointestinal tract
Organ:
liver
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Weight gain, efficiency of food utilization and sample intake of rats fed alkyl benzene for 12 weeksa. (Oser and Morgareidge, 1965)

Dosageb sex Mean weight gain (g) Efficiency of food utilization (g/100 g)c sample intake (g/kg/day)
Controls M 313 18.8 None
  F 169 13
0.05 g/kg/day M 315 18.6 0.05
  F 164 12.6
0.25 g/kg/day M 314 18.8 0.256
  F 163 12.6

a: The values given are the means of 7 or 8 animals of each sex.

b: These are nominal dosages. The actual dosages, calculated from food consumption, are given

as “sample intake” in the last column.

c: As grams of weight gained per 100 g of food eaten.

Table 2: Organ weights of rats fed alkyl benzene sulfonates in the diet for 12 weeks (Oser and Morgareidge, 1965)

Organ Sex Organ weighta(%)
Controls 0.05 g LAS/kg/day 0.25 g LAS/kg/day
Liver M 3.58 3.81 3.79
F 3.86 3.77 4.34b
Kidneys M 0.7 0.75 0.73
F 0.79 0.8 0.78
Spleen M 0.21 0.25 0.22
F 0.31 0.28 0.3
Heart M 0.36 0.33 0.34
F 0.39 0.4 0.37
Adrenals M 0.013 0.012 0.012
F 0.027 0.029 0.027
Pituitary M 0.004 0.004 0.004
F 0.007 0.006 0.007
Cecum M 0.47 0.49 0.5
F 0.58 0.57 0.58

a: as percentage of bodyweight.

b: Different from corresponding controls of same sex, P < 0.01.

Table3: Gross pathologic findings in rat fed alkyl benzene sulfonates in the diet for 12 weeksa (Oser and Morgareidge, 1965)

Organ and findings Control 0.05 g LAS/kg/day 0.25 g LAS/kg/day
M F M F M F
Cecum: Slight hyperemia - - - 5112 - -
Kidneys: Soft and hollow - - 5098 5106; 5111 5124 5146; 5148
Hollow and filled with fluid - 5022 - 5113 - 5147
Congenital absence of right kidney left; hypertrophied 5009 - - - - -
Adrenals: Hyperemic and/or slightly hemorrhagic 5003;5012 -   5114; 5119 5124; 5125 5145
Thymus: Petecbial hemorrhage - - - 5113 5130 -
Lungs: Hemorrhagic areas 5002 5029 5092 5114; 5115; 5119 5124; 5130 5144; 5150
Purulent leisions - - 5102 - - -
Edema - - - - - 5136
Areas of emphysema - 5030 5091 5110 - -

a: The numbers given are the identifying numbers of the individual animals that exhibited the lesion.

Conclusions:
Administration of sodium linear alkylbenzene sulphonate with nominal chain length of C12 (range C9-15) to albino rats at dose levels of 0.05 and 0.25 g/kg bw/day resulted in a NOAEL of 0.05 g/kg bw/day for female rats, based on increased relative liver weights and 0.25 g/kg bw/day for male rats, based on no abnormal variations in growth, food intake, hematology, clinical biochemistry, urinalysis, gross pathology, organ weights and histopathology in any of the groups observed
Executive summary:

A sub-chronic repeated dose dietary toxicity study was conducted with C10-14 LAS, sodium salt in Wistar FDRL rats. The test substance was administered to groups of 5 male and 5 female rats at dietary dose levels of 0, 50 and 250 mg/kg bw/day. Appearance, behaviour and overt signs of toxicity were observed daily. Food consumption and body weights of the rats of were recorded once weekly. During 6th and 12th week on test haematological, clinical chemistry and urine parameters were analysed. Following the 12-week clinical examinations, all surviving rats were sacrificed and gross pathological examination was performed. The major organs were weighed and histopathology was performed. No adverse effects on body weight gain, food consumption and behaviour were observed. The clinical data showed no abnormal variations in any of the dose groups. The relative organ weights and the histopathological evaluation did not show significant differences among the dose groups except for a significant liver weight increase in females of the highest dose group. Under the study conditions, the systemic toxicity NOAEL was established at 50 mg/kg bw/day based on increased relative liver weights in the females at 250 mg/kg bw/day (Oser, 1965).

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Principles of method if other than guideline:
The purpose of this study is to determine the chronic toxicity (oral) of C10 - C13 linear alkylbenzene sulfonic acid magnesium salt (LAS-Mg) in CRJ-SD rats. Male and female rats were administered with 0, 75, 150 and 300 mg/kg bw/day of test substance for 26 weeks. General symptoms and body weight of animals were recorded on a daily-basis and food intake were recorded weekly. Urinalysis and hematological examinations were performed in week 5 and 13 of the study. At the end of study, urinalysis, haematological and serum biochemistry were also done. All the surviving animals were humanely euthanized after 26 weeks and organs (liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary, brain and thyroid) were collected for recording absolute and relative organ weights. All the collected organs (including stomach, duodenum, ileum, cecum, pancreas, bone marrow) were examined microscopically.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: CRJ-SD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female rats were obtained from Charles River Laboratories Japan, Inc.
- Age at study initiation: Approximately 4 weeks
- Weight at study initiation: Not reported
- Fasting period before study: Not reported
- Housing: Animals were housed in cages with 2 animals/cage.
- Diet: Solid feed (CE-2, CLEA Japan, Inc.); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1°C
- Humidity: 55 ± 5%
- Air changes: Not reported
- Photoperiod: Not reported

IN LIFE DATES: Not reported
Route of administration:
oral: gavage
Details on route of administration:
Test substance was forcibly administered using a metallic gastric sonde, once daily
Vehicle:
water
Remarks:
(distilled)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Test substance solutions were prepared in distilled water to prepare required concentrations of 75, 150 and 300 mg/kg bw/day
- Storage conditions: Not reported
- DOSE VOLUME: 3 mL/kg bw for test substance as well as control group
- Preparation (%): 0, 2.5, 5 and 10% respectively for administration of doses of 0, 75, 150 and 300 mg/kg bw/day
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
26 weeks
Frequency of treatment:
Once daily for 26 weeks (excluding Sundays)
Dose / conc.:
75 mg/kg bw/day (actual dose received)
Dose / conc.:
150 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
20 animals/sex/dose group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected based on the subacute toxicity study of LAS-Mg, in which there was slight weight suppression of animals at 620 mg/kg bw/day (1/3 LD50). Hence, the high dose for this study was selected as 300 mg/kg (1/6 LD50). The lower doses were set at a common ratio of 2, with the medium dose being 150 mg/kg (1/12 LD50) and the low dose being 75 mg/kg (1/24 LD50).
- Rationale for animal assignment: Not reported
- Rationale for selecting satellite groups: No satellite groups were used in the study
Positive control:
Not included
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: All animals were observed daily for general symptoms.

BODY WEIGHT: Yes
- Time schedule for examinations: All animals were weighed daily.

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule: Amount of food intake was measured once per week.

FOOD EFFICIENCY: Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During weeks 5, 11 and 26
- Anaesthetic used for blood collection: Yes. The animals were anesthetized with sodium pentobarbital
- Animals fasted: Not reported
- How many animals: During weeks 5 and 11, blood was collected from 10 animals/sex from the control group and 300 mg/kg bw/day group and in week 26, blood was collected from all animals.
- Parameters checked: Red blood cell count, white blood cell count, hemoglobin level and hematocrit level were measured, and white blood cell fractions were calculated by Giemsa-staining the blood smear samples.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At end of study i.e. 26 weeks
- Animals fasted: Not reported
- How many animals: All animals
- Parameters checked: s-GOT, s-GPT, alkaline phosphatase, total protein, albumin, glucose, cholesterol, bilirubin, creatinine, urea nitrogen, inorganic phosphates and Cl-, Na+, K+, Ca2+ and Mg2+

URINALYSIS: Yes
- Time schedule for collection of urine: During weeks 5, 11 and 26
- Metabolism cages used for collection of urine: No
- Animals fasted: Not reported
- How many animals: During weeks 5 and 11, urine was collected from 10 animals/sex from the control group and 300 mg/kg bw/day group and in week 26, urine was collected from all animals.
- Method: Lab sticks, Urobilistix and iktstix method (Semi-quantitative investigation)
- Parameters: pH, protein, glucose, ketone bodies, occult blood, urobilinogen and bilirubin

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals were euthanized after blood sampling and organs were immediately collected.

TISSUE COLLECTED FOR ORGAN WEIGHTS AND HISTOPATHOLOGY: Liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary, brain and thyroid

HISTOPATHOLOGY: Above collected organs and stomach, duodenum, ileum, cecum, pancreas, bone marrow were fixed in 10% formalin, embedded in paraffin and underwent H&E staining before being examined under the microscope
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
There were wetting in the area surrounding animals’ mouths after dosing, and a few animals had abnormal airway sounds. Soft stools were seen in animals after one month, then recovered in few days.
Mortality:
mortality observed, treatment-related
Description (incidence):
One of the male animals in the 300 mg/kg bw/day dose group died (in week 21) from weakness due to significant weight loss involving diarrhea and loss of appetite.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was slight body weight suppression in the male animals of 300 mg/kg bw/day dose group.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
5 weeks interim analysis: There were decrease in segmented neutrophils and increase in lymphocytes in male animals in the 300 mg/kg bw/day dose group
13 weeks interim analysis: Significant decrease in basophils and hematocrit levels was observed for females and significant increase in segmented neutrophils and white blood cell count and significant decrease in lymphocytes were observed for males the 300 mg/kg bw/day dose group.
6 month analysis: Significant reduction in hematocrit levels for female animals in 300 mg/kg bw/day dose group was the only effect seen in hematological parameters.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
In 300 mg/kg bw/day dose groups: There was significant reduction in bilirubin, protein, albumin and calcium levels in male animals and in s-GPT, glucose, calcium and magnesium levels in female animals
In 150 mg/kg bw/day dose groups: There was significant reduction in s-GOT, s-GPT, alkaline phosphatase, protein, albumin, sodium and calcium levels in male animals and in chloride levels in female animals.
No effects were observed in 75 mg/kg bw/day treated animals.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
5 weeks interim analysis: No significant effects (on protein, occult blood, Urobilinogen and pH) were observed in treated animals as compare to control animals
13 weeks interim analysis: Male animals in the 300 mg/kg bw/day dose group showed + protein results. Others parameters (occult blood, Urobilinogen and pH) were comparable in control and treated animals.
26 weeks analysis: There were changes in protein levels in treated male animals as compare to control groups.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In 300 mg/kg bw/day dose groups: Significant increased liver weights (absolute weight in females and relative weight in males), actual and relative thymus weights (males) and relative pituitary (males) weights were observed in treated animals as compare to control. Reduction in actual and relative heart weights (females), relative kidneys weights (males), and relative adrenal gland weights (females) were also observed in highest dose treated group.

In 150 mg/kg bw/day dose groups: Significant increased actual and relative thymus weights and actual pituitary weights were observed in male animals as compare to control. There was decreased relative heart weights in females and relative kidneys weights in males of this dose group.

In 75 mg/kg bw/day dose group: Significant dip in absolute heart weights and relative adrenal glands weights of females was seen in low dose group.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- Yellow nodules was observed on liver for one male animal in the 75 mg/kg group.
- White spots were observed on adrenal glands for one female animal in the 75 mg/kg group.
- Bleeding spots in stomach were observed for two female animals in the 300 mg/kg group and one male animal in the 75 mg/kg group. Erosion was observed for one male animal in the 150 mg/kg group. Thickening of the proventriculus and yellow crust formation on the mucosal surface were observed for 11 male animals in the 300 mg/kg group and one male animal in the 150 mg/kg group.
- Stasis and diffuse emphysema in lungs were observed for one male animal in the control group.
- Edema was observed in pituitary for one male animal in the 150 mg/kg group.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Control group:
- Hyperprexia of the bile duct was observed for one female, while steatosis was observed for another female animal.
- Renal tubular hyaline casts were observed for two each of male and female animals.
- Fibrosis of islets of Langerhans observed for two male animals.
- Stasis was observed for one male animal.
- Interstitial myocarditis was observed for two male animals, while epicarditis was observed for another animal.
300 mg/kg bw/day dose group:
- Interstitial small round cell infiltration was observed in kidney for one male animal and renal tubular calcification was observed for one female animal.
- Small round cell infiltration in the submucosal layer of stomach was observed for one male animal.

150 mg/kg bw/day dose group:
- Kupffer cell migration was observed for one male animal.
- Renal tubular hyaline casts were observed for one male animal, interstitial small round cell infiltration was seen for two male animals, renal tubular epithelial cell swelling was observed for one male animal and renal tubular calcification was observed for one female animal.
- Interstitial myocarditis was observed for two male animals.

75 mg/kg bw/day dose group:
- Focal necrosis was observed for one male animal, while Kupffer cell migration and sinusoidal dilation were observed for one female animal.
- Renal tubular hyaline casts were observed for three male animals, while interstitial small round cell infiltration was observed for two other male animals.
- Fibrosis of islets of Langerhans was observed for one male animal, and mild bleeding in the peripancreatic lymph glands were observed for another male animal.
-Interstitial pneumonia was observed for one female animal.
- Although interstitial myocarditis was observed for five male animals, one of them was a moderate change. Another animal experienced epicarditis.
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
- There was animals wetting in the area surrounding their mouths after dosing, and a few animals had abnormal airway sounds. Soft stools were seen in animals after one month, then recovered in few days.

- One of the male animals in the 300 mg/kg bw/day dose group died (in week 21) from weakness due to significant weight loss involving diarrhea and loss of appetite.

BODY WEIGHT AND WEIGHT GAIN
- There was slight body weight suppression in the male animals of 300 mg/kg bw/day dose group.

FOOD CONSUMPTION AND COMPOUND INTAKE
- No significant difference was noted in males or females between the control and test substance groups.

FOOD EFFICIENCY
- No effects were observed in test substance treated animals as compare to control animals

HAEMATOLOGY
- 5 weeks interim analysis: There were decrease in segmented neutrophils and increase in lymphocytes in male animals in the 300 mg/kg bw/day dose group
- 13 weeks interim analysis: level: Significant decrease in basophils and hematocrit levels was observed for females and significant increase in segmented neutrophils and white blood cell count and signicant decrease in lymphocytes were observed for males the 300 mg/kg bw/day dose group.
- 6 month analysis: Significant reduction in hematocrit levels for female animals in 300 mg/kg bw/day dose group was the only effect seen in hematological parameters.

CLINICAL CHEMISTRY
- In 300 mg/kg bw/day dose groups: There was significant reduction in bilirubin, protein, albumin and calcium levels in male animals and significant reduction in s-GPT, glucose, calcium and magnesium levels in female animals
- In 150 mg/kg bw/day dose groups: There was significant reduction in s-GOT, s-GPT, alkaline phosphatase, protein, albumin, sodium and calcium levels in male animals and significant reduction in chloride levels in female animals.
- No effects were observed in 75 mg/kg bw/day treated animals.

URINALYSIS
- 5 weeks interim analysis: No significant effects (on protein, occult blood, Urobilinogen and pH) were observed in treated animals as compare to control animals
- 13 weeks interim analysis: Male animals in the 300 mg/kg bw/day dose group showed + protein results. Others parameters (occult blood, Urobilinogen and pH) were comparable in control and treated animals.
- 26 weeks analysis: There were changes in protein levels in treated male animals as compare to control groups.

ORGAN WEIGHTS
- In 300 mg/kg bw/day dose groups: Significant increased liver weights (absolute weight in females and relative weight in males), actual and relative thymus weights (males) and relative pituitary (males) weights were observed in treated animals as compare to control. Reduction in actual and relative heart weights (females), relative kidneys weights (males), and relative adrenal gland weights (females) were also observed in highest dose treated group.

- In 150 mg/kg bw/day dose groups: Significant increased actual and relative thymus weights and actual pituitary weights were observed in male animals as compare to control. There was decreased relative heart weights in females and relative kidneys weights in males of this dose group.

- In 75 mg/kg bw/day dose group: Significant dip in absolute heart weights and relative adrenal glands weights of females was seen in low dose group.

GROSS PATHOLOGY
- Yellow nodules was observed on liver for one male animal in the 75 mg/kg group.
- White spots were observed on adrenal glands for one female animal in the 75 mg/kg group.
- Bleeding spots in stomach were observed for two female animals in the 300 mg/kg group and one male animal in the 75 mg/kg group. Erosion was observed for one male animal in the 150 mg/kg group. Thickening of the proventriculus and yellow crust formation on the mucosal surface were observed for 11 male animals in the 300 mg/kg group and one male animal in the 150 mg/kg group.
- Stasis and diffuse emphysema in lungs were observed for one male animal in the control group.
- Edema was observed in pituitary for one male animal in the 150 mg/kg group.

HISTOPATHOLOGY
Control group:
- Hyperprexia of the bile duct was observed for one female, while steatosis was observed for another female animal.
- Renal tubular hyaline casts were observed for two each of male and female animals.
- Fibrosis of islets of Langerhans observed for two male animals.
- Stasis was observed for one male animal.
- Interstitial myocarditis was observed for two male animals, while epicarditis was observed for another animal.
300 mg/kg bw/day dose group:
- Interstitial small round cell infiltration was observed in kidney for one male animal and renal tubular calcification was observed for one female animal.
- Small round cell infiltration in the submucosal layer of stomach was observed for one male animal.

150 mg/kg bw/day dose group:
- Kupffer cell migration was observed for one male animal.
- Renal tubular hyaline casts were observed for one male animal, interstitial small round cell infiltration was seen for two male animals, renal tubular epithelial cell swelling was observed for one male animal and renal tubular calcification was observed for one female animal.
- Interstitial myocarditis was observed for two male animals.

75 mg/kg bw/day dose group:
- Focal necrosis was observed for one male animal, while Kupffer cell migration and sinusoidal dilation were observed for one female animal.
- Renal tubular hyaline casts were observed for three male animals, while interstitial small round cell infiltration was observed for two other male animals.
- Fibrosis of islets of Langerhans was observed for one male animal, and mild bleeding in the peripancreatic lymph glands were observed for another male animal.
-Interstitial pneumonia was observed for one female animal.
- Although interstitial myocarditis was observed for five male animals, one of them was a moderate change. Another animal experienced epicarditis.
Key result
Dose descriptor:
NOAEL
Effect level:
150 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
histopathology: non-neoplastic
Remarks on result:
other:
Remarks:
Based on body weight gain suppression, mortality, fluctuations in hematological and serum biochemistry, altered organ weights and histopathological alterations in kidney at 300 mg/kg bw/day.
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
300 mg/kg bw/day (actual dose received)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes
Conclusions:
Oral administration of C10-13 magnesium linear alkylbenzene sulphonate (LAS-Mg; purity: 96.9%) to CRJ-SD rats at dose levels of 0, 75, 150 and 300 kg/kg bw/day for 26 weeks revealed no observed adverse effect level (NOAEL) of 150 mg/kg bw/day, based on weight suppression, mortality, fluctuations in hematological and serum biochemistry, altered organ weights and histopathological alterations in kidney at 300 mg/kg bw/day.
Executive summary:

A sub-chronic repeated dose toxicity study was conducted with C10-13 LAS, magnesium salt in CRJ-SD rats. The test substance was administered daily at dose levels of 0, 75, 150 and 300 mg/kg bw/day via oral gavage to groups of 20 animals/sex for 26 weeks. General symptoms and body weight of animals were recorded on a daily basis and food intake was recorded weekly. Urinalysis, haematological and clinical biochemistry examinations were performed during the in-life phase as well as at termination. Following necropsy of the surviving animals, gross pathological and histopathological examinations were performed. There was slight body weight suppression in the males at 300 mg/kg bw/day. Haematological findings revealed significant reduction in haematocrit levels for females at 300 mg/kg bw/day. There was significant reduction in bilirubin, protein, albumin and calcium levels in male animals and significant reduction in SGPT, glucose, calcium and magnesium levels in female animals at 300 mg/kg bw/day. Reduction in relative heart weight (females), kidney weight (males) and adrenal gland weight (females) were also observed in the highest dose group. In the kidneys, renal tubular epithelial cell swelling, hyaline casts and interstitial small round cell infiltration were observed at 300 mg/kg bw/day. There were no toxicologically significant effects at the lower doses. Under the study conditions, the systemic toxicity NOAEL was established at 150 mg/kg bw/day, based on reduction in body weight, fluctuations in haematological and serum biochemistry, altered organ weights and histopathological alterations in kidney at 300 mg/kg bw/day (Ito, 1978b).

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Principles of method if other than guideline:
The purpose of this study is to determine the sub-acute toxicity of C10-13 Magnesium Linear Alkyl Benzene Sulfonate Salts (LAS-Mg) in CRJ-SD rats. LAS-Mg was orally administered at 0 (distilled water), 155, 310 and 620 mg/kg bw to male and females rats for 29 and 30 days, respectively. General symptoms of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters. After collection of blood samples, the animals were humanely euthanized, and gross necropsy, organs weights and histopathology was performed.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: CRJ-SD rats
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc.
- Age at study initiation: 5 weeks
- Weight at study initiation: Not reported
- Fasting period before study: Not reported
- Housing: 3 male and 4 female animals per cage.
- Diet: Solid feed (CE-2, CLEA Japan, Inc.); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: After 1 week pre-housing, the animals were used in the experiment.

ENVIRONMENTAL CONDITIONS
- Temperature: 23 ± 2°C
- Humidity: 55 ± 5%
- Air changes: Not reported
- Photoperiod: Not reported

EXPERIMENTAL START DATE: Not reported
Route of administration:
oral: gavage
Details on route of administration:
The test substance was administered orally through a metallic gastric sonde.
Vehicle:
other: Distilled water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance was suspended in distilled water prior to dosing.

- DOSE VOLUME: 5 mL/kg bw

- Concentration in vehicle: 0, 310, 620, and 1240 mg/mL respectively for administration of doses of
0, 155, 310 and 620 mg/kg bw.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
29 days for males and 30 days for females
Frequency of treatment:
Once daily
Dose / conc.:
155 mg/kg bw/day (actual dose received)
Remarks:
In distilled water (corresponding to 310 mg/mL)
Dose / conc.:
310 mg/kg bw/day (actual dose received)
Remarks:
In distilled water (corresponding to 620 mg/mL)
Dose / conc.:
620 mg/kg bw/day (actual dose received)
Remarks:
In distilled water (corresponding to 1240 mg/mL)
No. of animals per sex per dose:
Test group: 12 animals/sex/dose group
Control group: 15 male/group and 16 female/group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels for this study were selected based on the results of acute toxicity study conducted with LAS-Mg in rats. 1/3 of the LD50 levels of LAS-Mg, i.e. 620 mg/kg bw, was selected as the highest dose. The other two doses were set at common ratios of 2, whereby the medium and low doses were 310 (1/6 LD50) and 155 (1/12 LD50) mg/kg bw, respectively.
Positive control:
No
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Daily

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Time schedule for examinations: Twice/week

FOOD EFFICIENCY: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected was collected from the carotid artery using a cannula.
- Anesthetic used for blood collection: Yes, light ether anesthesia was used.
- Animals fasted: Not reported
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red and white blood cell count were measured using an auto-analyzer SMA-4A.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 1 month, blood was collected from each animal and centrifuged (at 3000 rpm for 15 minutes) to obtain the serum samples for analysis.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: s-GOT, s-GPT, alkaline phosphatase, total protein, albumin, glucose, cholesterol, bilirubin, creatinine, urea nitrogen, inorganic phosphates and ions (Na+, K+, Ca2+, Mg2+ and Cl-)

URINALYSIS: Yes
- Time schedule: Not reported
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: pH, proteins, sugar, ketone bodies and occult blood

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

ORGAN WEIGHTS: The wet weight of liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary and the brain were measured. Based on this, the relative weights were calculated.

HISTOPATHOLOGY: Yes
Based on the results of the organ weight measurements, histopathology was performed only for the livers of male animals from the test and control groups. After Bouin fixing, paraffin embedding and H&E staining, the samples were examined under the microscope.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
At 620 mg/kg bw, animals exhibited the action of wetting the area around the mouth and nose, and rubbing it against the cage. Many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 155 and 310 mg/kg bw, however, the animals showed recovery at low and mid doses.
Mortality:
mortality observed, treatment-related
Description (incidence):
One male animal (Day 6) and two female animals (Day 7 and 12, respectively) died from the high dose group (i.e. 620 mg/kg bw). In each case, the animal died from weakness after significant loss of body weight involving diarrhea and loss of appetite, and the necropsy revealed congestion of the gastric mucosa, intestinal relaxation, ascites retention, and it also involved lung congestion.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At 620 mg/kg bw, males exhibited a body weight suppression on Day 2. Thereafter, the growth curve showed the same rate of increase as the control group, but the suppression was observed again after 3 weeks. In the end, body weight suppression was observed in animals (both males and females) belonging to the high dose group (i.e. 620 mg/kg bw).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Animals showed a dose dependent decline in the feed consumption.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Decreased feed efficacy was observed for male animals at 620 mg/kg bw.
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Decreased hemoglobin and hematocrit level was observed in the male animals of high dose group (i.e. 620 mg/kg bw). No changes were observed in the RBC or WBC count.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Decreased s-GOT (only males), K+ (only males) and Ca2+ levels (only females) was observed in the low dose group (i.e. 155 mg/kg bw). Increased s-GOT (both males and females) and alkaline phosphatase (only males), and decreased total proteins (only males), creatinine (only males) and Ca2+ levels (both males and females) was observed in the mid dose group (i.e. 310 mg/kg bw). Increased s-GOT, s-GPT and alkaline phosphatase (both males and females), and decreased total proteins (only females), albumin (only females), glucose (only females), cholesterol (only males) K+ (only males) and Ca2+ levels (both males and females) was observed in the high dose group (i.e. 620 mg/kg bw). The effects were not dose-dependent.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The relative liver weight of male and female animals was significantly increased at 620 mg/kg bw (p<0.01). Apart from this, increase in relative weight of testes (p<0.01) and brain (p<0.01) was also observed at 620 mg/kg bw. Decrease in the relative weight of kidney (both side at 310 mg/kg bw; right side at 155 mg/kg bw, p<0.05), adrenal gland (both sides at 155 and 310 mg/kg bw, p<0.05), heart (at 310 and 620 mg/kg bw, p<0.01) and thymus (at 620 mg/kg bw, p<0.05) was observed in the female animals. Although, some significant effects were observed at low and mid dose yet they were not dose-dependent.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
At 620 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Focal necrosis and increased glycogen levels were observed in the liver of few animals belonging to mid and low dose groups, but these findings were common with the control group and also showed no dose-dependency.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
CLINICAL SIGNS: At 620 mg/kg bw, many animals exhibited diarrhea from the start of dosing as well as a suppressed state, several hours after dosing. Soft stool was observed in some animals dosed at 155 and 310 mg/kg bw, however, the animals showed recovery at low and mid doses.

MORTALITY: One male animal (Day 6) and two female animals (Day 7 and 12, respectively) died from the high dose group (i.e. 620 mg/kg bw). In each case, the animal died from weakness after significant loss of body weight involving diarrhea and loss of appetite, and the necropsy revealed congestion of the gastric mucosa, intestinal relaxation, ascites retention, and it also involved lung congestion.

BODY WEIGHT AND WEIGHT CHANGES: Body weight suppression was observed in animals (both males and females) belonging to the high dose group (i.e. 620 mg/kg bw).

FOOD CONSUMPTION AND COMPOUND INTAKE: Animals showed a dose dependent decline in the feed consumption. Decreased feed efficacy was observed for male animals at 620 mg/kg bw.

HAEMATOLOGY: Decreased hemoglobin and hematocrit level was observed in the male animals of high dose group (i.e. 620 mg/kg bw). No changes were observed in the RBC or WBC count.

CLINICAL BIOCHEMISTRY: Alterations in the biochemical parameter were observed in the test animals. However, the effects were not dose dependent.

URINALYSIS: Results of urinalysis (pH, proteins, sugar, ketone bodies and occult blood) in the test animals were comparable to the control group.

ORGAN WEIGHT: The relative liver weight of male and female animals was significantly increased at 620 mg/kg bw (p<0.01). Apart from this, increase in relative weight of testes (p<0.01) and brain (p<0.01) was also observed at 620 mg/kg bw. Decrease in the relative weight of kidney (both side at 310 mg/kg bw; right side at 155 mg/kg bw, p<0.05), adrenal gland (both sides at 155 and 310 mg/kg bw, p<0.05), heart (at 310 and 620 mg/kg bw, p<0.01) and thymus (at 620 mg/kg bw, p<0.05) was observed in the female animals. Although, some significant effects were observed at low and mid dose yet they were not dose-dependent.

GROSS PATHOLOGY: At 620 mg/kg bw, the animals showed hypertrophy in the stomach wall of the proventriculus, as if the process of inflammation and recovery had repeated over and over. Therefore, formation of new capillaries was observed on the peritoneal side. There were no additional changes relative to the control group although, events of hydronephrosis, bleeding plaques on the thymus and pneumonia were observed including the control group.

HISTOPATHOLOGY: Focal necrosis and increased glycogen levels were observed in the liver of few animals belonging to mid and low dose groups, but these findings were common with the control group and also showed no dose-dependency.
Key result
Dose descriptor:
NOAEL
Effect level:
310 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
mortality
body weight and weight gain
organ weights and organ / body weight ratios
Remarks on result:
other: Based on mortality, reduced body weight and changes in organ weight (without biochemical or histopathological correlation). The LOAEL was 620 mg/kg bw.
Conclusions:
Oral administration of C10-13 magnesium linear alkylbenzene sulphonate (LAS-Mg) to CRJ-SD rats at 0, 155, 310 and 620 mg/kg bw for 1 month resulted in a NOAEL of 310 mg/kg bw, based on mortality, reduced body weight and changes in organ weight (without biochemical or histopathological correlation) at higher dose level. The LOAEL was 620 mg/kg bw.
Executive summary:

A sub-acute repeated dose toxicity study was conducted with C10-13 LAS, magnesium salt in CRJ-SD rats. The test substance was administered to groups of male and female rats via oral gavage at doses of 0, 155, 310 and 620 mg/kg bw/day for 29/30 days. Clinical signs of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters, following which the animals were humanely euthanized, gross necropsy was performed and the organs weights were determined for the major organs. Based on the results of organ weight measurements, histopathology was performed only for the livers of male animals from the test and control groups. 1 male and 2 female animals died at the highest dose. The relative liver weight of male and female animals was significantly increased at 620 mg/kg bw/day. Apart from this, increase in relative testes weight and brain weight were also observed at this dose. Decrease in the relative heart weight (at 310 and 620 mg/kg bw/day) and thymus (at 620 mg/kg bw/day) was observed in the female animals. Although, some significant effects were observed at low and mid doses yet they were not dose-dependent. Histopathology revealed focal necrosis and increased glycogen levels in the liver of few animals belonging to the 155 and 310 mg/kg bw dose groups but these findings were comparable to the control group and were not dose-dependent. Under the study conditions, the systemic toxicity NOAEL was established at 310 mg/kg bw/day, based on mortality, reduced body weight and changes in organ weight (without biochemical or histopathological correlation) at the higher dose level (Ito, 1978c). 

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Reason / purpose:
reference to same study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
(test was conducted on males against the guideline which recommends both sexes should be used for evaluation of toxicity)
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
Albino rats
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Not specified
- Females nulliparous and non-pregnant: No
- Age at study initiation: Not specified
- Weight at study initiation: 319 g (average)
- Fasting period before study: Not specified
- Housing: Five animals were housed per cage.
- Diet: Ground purina laboratory chow: ad libitum
- Water: Ad libitum
- Acclimation period: Not specified

No further information was available for food and water quality and environmental conditions
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS: Not applicable

- DIET PREPARATION
The compound was fed at 0.01, 0.1% and 1% in a diet of ground Purina Laboratory Chow. The diets were prepared using a food mixer and only one batch of diet was necessary. The control group was fed only control diet of ground Purina Laboratory Chow.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
32 days
Frequency of treatment:
daily
Dose / conc.:
0 other: %
Dose / conc.:
1 other: %
Dose / conc.:
0.1 other: %
Dose / conc.:
0.01 other: %
No. of animals per sex per dose:
10 male/dose level
Control animals:
yes, concurrent vehicle
Details on study design:
Animals were fed with test substance at dose levels of 0.01, 0.1 and 1.0% in a diet of ground
Purina Laboratory Chow for a period of 32 days.
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: On day 0, 4, 8, 14, 21, 27 and 32.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, group feed consumption was recorded.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not specified

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: On day 28 of test
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: 10 Animals of dose level group of 0 and 1%
- Parameters: Hemoglobin, hematocrit, white blood cell count and differential leukocyte counts were examined.

CLINICAL CHEMISTRY: No

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
On 32nd and 33rd day of test, all surviving rats were euthanized by over dosage with ether and body weight and weight of liver, kidneys, spleen, heart, lung, brain, and testis of each rat were recorded. Animals were then examined for evidence of gross pathologic changes.

HISTOPATHOLOGY: Yes
Representative specimens of trachea, lungs, heart, tongue, esophagus, stomach, small intestine, large intestine, liver kidneys, urinary bladder, pituitary, thyroid, adrenal, pancreas, testis, spleen, femoral bone marrow, cerebrum, cerebellum and eyes were preserved in neutral buffered 10% formalin for microscopic examination.
Statistics:
The organ weight was analyzed using analysis of variance test (p=0.05). All significant “F” values were further analyzed using “Duncan’s new multiple range test”.
Clinical signs:
no effects observed
Mortality:
not specified
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
The absolute as well as relative liver weights were significantly increased for two low dose levels (0.01 and 1%) as compare to controls. However, liver weights of the high dose group animal (1.0%) did not differ from the controls.
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
- No adverse effect of ethanolamine was observed for survival, body weight, food consumption, hematology, gross pathology and histopathology.

ORGAN WEIGHTS:
The absolute as well as relative liver weights were significantly increased for two low dose levels (0.01 and 1%) as compare to controls. However, liver weights of the high dose group animal (1.0%) did not differ from the controls. Since the observed liver weights changes were not dose dependent (only at lower dose levels, not at highest dose) and no changes were observed in liver’s gross pathology and histopathology observations. Therefore, this effect is not considered treatment related.
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 other: %
Based on:
act. ingr.
Sex:
male
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: no abnormal variations in any of the evaluated parameters i.e. body weight gain, feed efficiency, hematologic, organ weights, pathological and microscopic findings.

Table 1. Duncan's new multiple range test applied to significant "F" ratios in the analysis of variance. Any two means not underscored by the same line are significantly different (p=0.05) (Terhaar et.al. 1967)

Liver (wt. g.) Dose level control 1% 0.10% 0.01%
  Mean 18.64 20.47 21.32 22.66
Liver (wt. %) Dose level control 1% 0.10% 0.01%
  Mean 4.07 4.32 4.45 4.7
Conclusions:
Administration of ethanolamine to albino Sprague dawley rats at dose levels of 0, 0.01%, 0.1% and 1% resulted in no abnormal variations in any of the evaluated parameters i.e. body weight gain, feed efficiency, hematologic, organ weights, pathological and microscopic findings. The NOAEL was therefore concluded at >1%.
Executive summary:

 A sub-acute repeated dose toxicity study was conducted with monoethanolamine (MEA) in Sprague-Dawley rats. The test substance was administered to the rats through diet at dose levels of 0%, 0.01%, 0.1% and 1% (equivalent to 0, 7.7, 77 and 770 mg/kg bw/day) for a period of 32 days during which individual body weight and group feed consumption were recorded. On Day 28, haematological examination was performed on the control and high dose group animals. Following sacrifice of the surviving animals, gross pathology of all organs and weight of liver, kidneys, spleen, heart, lung, brain, and testes were recorded. No significant difference in body weight and feed consumption were observed. There was also no difference found in the mean haematogram of the high dose rats. No treatment-related effects were observed in the organ weights, gross pathology and histopathology of the organs. Under the study conditions, the systemic toxicity NOAEL was established at dietary level of 1% based on absence of effects in body weight gain, feed efficiency, hematologic, organ weights, pathological and microscopic findings (Terhaar, 1967).

Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1974
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
A composite of dyes and base components found in semi-permanent hair color products was administered in the diet to dogs for 2 years to assess chronic toxicity of the composite. Male and female pure-bred beagle dogs were exposed to 0.0, 19.5, and 97.5 mg/kg bw/day of composite in diet. Each animal was observed for physical, clinical, hematological, blood chemical and urinalysis parameters at different time intervals during the study. Necropsy was performed on one male and one female from each group at 6, 12, and 18 months, and on all survivors at 24 months. Organ weights and histopathology were performed after euthanizing the animals.
GLP compliance:
no
Limit test:
no
Species:
dog
Strain:
Beagle
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Beagles for research, White sulfur springs, New York
- Age at study initiation: 6-8 months
- Fasting period before study: No data
- Housing: Animals were housed individually
- Diet: Ralston Purina Dog Meal, Ralston Purina Company, St. Louis, Missouri; ad libitum
- Water: ad libitum
- Acclimation period: 3 weeks
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet: Diets were prepared fresh 7 days per week and were usually consumed within 1 hour after feeding each day.
- Diet: Ralston Purina Dog Meal, Ralston Purina Company, St. Louis Missouri.
- Adjustments of the concentration of test substance in the diet were made as required by weekly body weight changes.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
2 years
Frequency of treatment:
daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
19.5 mg/kg bw/day (nominal)
Dose / conc.:
97.5 mg/kg bw/day (nominal)
No. of animals per sex per dose:
2 years
Control animals:
yes, plain diet
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Twice for each dog

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: Daily

FOOD EFFICIENCY: Not specified

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day 0 and at 3, 6, 12, 18, 24 months
- Anesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: Hematology were performed in all dogs in the high dose and control groups and on three male and three female dogs in the low dose group.
- Parameters examined: Total and differential leucocyte counts, hematocrit, hemoglobin concentration, erythrocyte sedimentation rate, and prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Day 0 and at 3, 6, 12, 18, 24 months
- Animals fasted: Yes, 18 hours
- Parameters checked: Serum glucose, blood urea nitrogen, creatinine, uric acid, alkaline phosphatase and serum glutamic pyruvic transaminase activities

URINALYSIS: Yes
- Time schedule for collection of urine: Day 0 and at 3, 6, 12, 18, 24 months
- Metabolism cages used for collection of urine: No data
- Animals fasted:
- Parameters examined: Qualitative tests for the detection of occult blood, albumin, glucose, pH, and microscopic examination of urinary sediment.

OPTHAMOLOGY: Yes
- Time schedule for examination: once

NEUROBEHAVIOURAL EXAMINATION: Not examined

IMMUNOLOGY: Not examined


Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Time schedule: Necropsy was performed on one male and one female from each group at 6, 12, and 18 months, and on all survivors at 24 months

HISTOPATHOLOGY: Yes
Individual organ weights of the pituitary, adrenals, thyroid, liver, spleen, kidneys, and gonads from each animal were recorded and organ to body weight ratios calculated. Sections of sternum, rib, thymus, heart, aorta, lungs, eyes, salivary gland, lymph nodes, thyroid, trachea , esophagus, liver, gall bladder, spleen, pancreas, stomach, small intestine (three sections), large intestine, adrenals, kidneys, gonads, uterus, mammary gland, urinary bladder, gastrocnemius muscle, sciatic nerve, femur, spinal cord, brain, pituitary, and abdominal skin were fixed in formalin and stained with hematoxylin and eosin for subsequent microscopic examination. In addition, sections of liver and urinary bladder taken from all 18 dogs on whom a necropsy was performed at 24 months were fixed in paraformaldehyde solution for subsequent electron microscopic evaluation.
Other examinations:
Physical examinations including funduscopic, EKG, blood pressure, pulse rate and body temperature determinations were conducted initially and at 3, 6, 12, 18, and 24 months.
Statistics:
The data were subjected to statistical analysis, using the 95% confidence level. The methods used included chi square test, analysis of variance and t test, and the Fisher exact probability test.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
all dogs gained weight normally
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
Hematological values for male and female test and control animals were within normal limits. Hemoglobin and hematocrit values increased slightly in dogs from all groups in the study, reflecting normal maturation processes
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Blood values for serum glucose, uric acid, and creatinine concentrations, and for alkaline phosphatase and serum glutamic-pyruvic transaminase activities were within normal ranges for dogs from all groups. The concentrations of BUN usually were within normal ranges except for two control female dogs and one low dosage male at the 12-months test interval. At this time, these dogs showed BUN values of 40-45mg/100 mL. However, at 18 months, the BUN values for these three dogs were in the range of 15-25 mg/100 mL.
Urinalysis findings:
effects observed, non-treatment-related
Description (incidence and severity):
All animals in both test groups excreted blue-brown colored urine daily. However, urine analyses showed no remarkable findings at any examination period. Color was normal in the urines collected following overnight fasting and was probably an indication of rapid clearance.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
- Mortality: No mortality was observed throughout the study.
- No adverse effects were observed for clinical signs, body weight change, food consumption, organ weights gross pathology and histopathology

HEMATOLOGY: Hematological values for male and female test and control animals were within normal limits. Hemoglobin and hematocrit values increased slightly in dogs from all groups in the study, reflecting normal maturation processes.

CLINICAL BIOCHEMISTRY: Blood values for serum glucose, uric acid, and creatinine concentrations, and for alkaline phosphatase and serum glutamic-pyruvic transaminase activities were within normal ranges for dogs from all groups. The concentrations of BUN usually were within normal ranges except for two control female dogs and one low dosage male at the 12 months test interval. At this time, these dogs showed BUN values of 40-45 mg/100 mL. However, at 18 months, the BUN values for these three dogs were in the range of 15-25 mg/100 mL.

URINALYSIS: All animals in both test groups excreted blue-brown colored urine daily. However, urine analyses showed no remarkable findings at any examination period. Color was normal in the urines collected following overnight fasting and was probably an indication of rapid clearance.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 97.5 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No adverse effect was observed on any of the observed parameters (clinical signs, mortality, body weight, food consumption, haematology, clinical biochemistry, urinalysis, organ weights, gross pathology and histopathology)
Conclusions:
Administration of a composite of dyes and base components containing 22.42% monoethanolamine to dogs at dose levels of 0, 19.5 and 97.5 mg/kg bw/day resulted in no adverse effect on any of the observed parameters (clinical signs, mortality, body weight, food consumption, hematology, clinical biochemistry, urinalysis, organ weights, gross pathology and histopathology). Therefore, chronic feeding of composite (containing 22.42% monoethanolamine) has no impact on dogs up to ≥97.5 mg/kg bw/day (= 20.5 mg a.i. monoethanolamine/kg bw/day).
Executive summary:

A chronic repeated dose toxicity was conducted with monoethanolamine (MEA) in Beagle dogs. The test substance as a hair dye composite (containing 22.42% MEA) was administered in the diet to groups of 3 male and 3 female dogs for 2 years at concentrations of 0, 19.5, and 97.5 mg/kg bw/day. Apart from regular observation of clinical signs of toxicity, body weight and feed consumption, physical examinations including funduscopic, EKG, blood pressure, pulse rate, and body temperature determinations were conducted initially and at 3, 6, 12, 18, and 24 months. Haematological, clinical biochemistry and urinalysis parameters were determined for the high dose and control groups and on three male and three female dogs in the low dose group at the same times. Necropsy was performed on one male and one female from each group at 6,12, and 18 months, and on all survivors at 24 months and organ weight determination, gross pathology and histopathology were performed. No treatment-related adverse effects were observed in any of the parameter studied between controls and animals at 19.5 or 97.5 mg/kg bw/day. Under the study conditions, the systemic toxicity NOAEL was established at ≥97.5 mg/kg bw/day (equivalent to 20.5 mg a.i. MEA/kg bw/day), based on absence of statistically significant effects in the treated animals (Wernick, 1975).

Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1972
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
A justification of the read-across approach is appended in Section 13.
Reason / purpose:
reference to same study
Reason / purpose:
read-across source
Principles of method if other than guideline:
The purpose of this study is to determine the sub chronic toxicity of C10 - C14 linear alkylbenzene sulfonic acid sodium salt (LAS) in Wistar SLC rats. Male and female rats were maintained on diets of 0, 0.07, 0.2, 0.6 and 1.8% LAS (equivalent to 0, 40, 115, 340, 1030 mg/kg bw/day) for 26 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, blood was collected for estimation of hematological and clinical chemistry parameters. Urinalysis was also performed 2 weeks before necropsy. All the surviving animals were humanely euthanized and gross necropsy, organs weights and histopathology were performed.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: Wistar SLC
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female were obtained from Shizuoka Agricultural Cooperative Association for Laboratory Animals
- Age at study initiation: Approximately 4 weeks
- Weight at study initiation: 80 - 90 g (male rats), 65 - 80 g (female rats)
- Fasting period before study: No
- Housing: 5 animals were housed per stainless steel cage.
- Diet: FII food (from Funabashi Farm); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: Not reported

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 55 - 65%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: From: May 30, 1972 To: Nov. 27, 1972
Route of administration:
oral: feed
Details on route of administration:
C10 - C14 LAS was administered to animals by mixing 0.07, 0.2, 0.6 and 1.8% with pulverized FII feed, the powders were shaped into solid form before being freely provided for feeding along with tap water. As control, solid FII feed was provided to the animals.
Vehicle:
other: C10 - C14 LAS was administered in diet
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
26 weeks
Frequency of treatment:
Continuous in diet (ad libitum)
Dose / conc.:
0.07 other: %
Remarks:
in diet
Dose / conc.:
0.2 other: %
Remarks:
in diet
Dose / conc.:
0.6 other: %
Remarks:
in diet
Dose / conc.:
1.8 other: %
Remarks:
in diet
Dose / conc.:
40 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.07% dose level)
Dose / conc.:
115 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.2% dose level)
Dose / conc.:
340 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.6% dose level)
Dose / conc.:
1 030 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 1.8% dose level)
No. of animals per sex per dose:
10 animals/sex/dose group
Control animals:
yes, plain diet
Positive control:
No
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for examinations: Weekly

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: Weekly

OPHTHALMOSCOPIC EXAMINATION: No

HEMATOLOGY: Yes
- Time schedule for collection of blood: After 26 weeks, blood was collected from each animal prior to autopsy (for hemoglobin, hematocrit, red blood cells, white blood fractions and platelets) and during euthanasia (erythrocyte membrane resistance)
- Anesthetic used for blood collection: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Hemoglobin, hematocrit, red blood cells, white blood fractions and platelets, erythrocyte membrane resistance and blood coagulation time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 26 weeks, blood was collected from each animal after euthanasia.
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Total proteins, GOT activity, ALP activity, LAP activity, urea nitrogen, A/G ratio and bilirubin

URINALYSIS: Yes
- Time schedule: 2 weeks prior to autopsy, urine isolated and collected from waste was analyzed.
- Metabolism cages used for collection of urine: No
- Animals fasted: No
- How many animals: All surviving animals
- Parameters: Proteins, glucose, ketone bodies, occult blood, bilirubin and urobilinogen

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Gross findings were obtained after euthanizing animals and macroscopic findings were observed.

ORGAN WEIGHTS: Brain, pituitary, thyroid, thymus, heart, lungs, liver, kidneys, spleen, adrenal gland, prostate, testes, uterus, ovaries and appendix

HISTOPATHOLOGY: Yes
Collected tissues along with stomach, large and small intestines, pancreas, epididymis, skin and bones) were weighed, examined and subjected to microscopy through H.E., PAS, lipid and silver staining.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Severe diarrhea was observed in the high concentration group (1.8%).
Mortality:
mortality observed, treatment-related
Description (incidence):
In 24th week, 1 male animal from the 1.8% dose group died.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was significant inhibition of weight gain in both males and females of 1.8% dose group.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Food consumption was low in the 1.8% dose group relative to the control.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
Water consumption was slightly low in animals of 1.8% dose group relative to the control
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
In females of 1.8% dose group, a significant decrease in hematocrit and hemoglobin and significant increase in the erythrocyte membrane resistance was observed.. For both males and female species, there was increase in white blood cell count at 1.8% dose.

Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
There was dose dependent significant increase in ALP levels (at mid and high dose) and significant decrease in total protein (at high dose).
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
1.8% dose group: Significant increase in relative weights of liver, suprarenal gland, testis, brain and caecum were observed in males and relative weights of heart, thyroid, liver, uterus, brain and caecum of females.
0.6% dose group: Significant increase in caecum relative weights in males and hypophysis in females.
0.2% dose group: Caecum relative weights were significantly increased in males and kidney relative weights were significantly decreased in females.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Color of the liver in 2 males and 5 females from the 1.8% group was lighter than normal and seemed to have a yellowish white color mixed to it. In addition, lung tumors were also seen in one female rat from the 0.2% group.

Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
The main findings were concerned with kidneys, liver and intestinal tract.
Kidney damage was seen at and above 0.2% dose groups. Mild hepatocyte changes were observed in animals of 1.8% dose group and microscopic changes in intestinal tract were observed in female rats of the 1.8 and 0.6% dose groups.
Histopathological findings: neoplastic:
not examined
Other effects:
not examined
Details on results:
CLINICAL SIGNS:
- Severe diarrhea was observed in animals in the high concentration groups (1.8%) immediately after starting the study, and although the diarrhea gradually became milder after about 1 week, the animal s did not recover fully by the end of the study. The animals moved slowly, and their lower abdomen was dirty.

MORTALITY: During week 24, 1 male animal from 1.8% dose group died.
BODY WEIGHT AND WEIGHT CHANGES: There was significant inhibition of weight gain in both males and females of 1.8% dose group, while the 0.6% dose groups showed slight but not significant trends to suppress weight gain.

FOOD CONSUMPTION AND COMPOUND INTAKE: Food consumption was low in the 1.8% dose group relative to the control.

WATER CONSUMPTION AND COMPOUND INTAKE: Water consumption was slightly low in the
1.8% dose group.

HEMATOLOGY:
- A significant decrease in hematocrit and hemoglobin, a significant increase in the erythrocyte membrane resistance was observed among female animals in the 1.8% dose groups.
- Male and female rats in 1.8% dose group showed increase in white blood cell count, increase in neutrophil fraction out of the white blood cells and a decrease in lymphocytes.

CLINICAL BIOCHEMISTRY: There was dose dependent increase in ALP levels (at 0.6 and 1.8% dose groups) and decrease in total protein (at 1.8% dose group)

URINALYSIS: No effects were observed.

ORGAN WEIGHT:
- In highest concentration group (1.8%), significant increase in relative weights of liver, suprarenal gland, testis, brain and caecum were observed in males and relative weights of heart, thyroid, liver, uterus, brain and caecum of females.In 0.6% dose group, there was significant increase in caecum relative weights in males and hypophysis in females.

In 0.2% dose group, caecum relative weights were significantly increased in males and kidney relative weights were significantly decreased in females.

GROSS PATHOLOGY
- The color of the liver in 2 males and 5 females from the 1.8% dose group was lighter than normal and seemed to have a yellowish white color mixed to it.
- In addition, lung tumors were seen in one female rat from the 0.2% dose group.

- Autopsy of one male rat from the LAS-1.8% dose group that died during the study had significant s welling of the abdomen. This was due to the swelling of the stomach and small intestines. The contents from the stomach to the first part of the jejunum was a blackish purple liquid. From there on, the ileum was filled with a semi-transparent, viscous, gelatinous substance.

HISTOPATHOLOGY

The main findings were concerned with kidneys, liver and intestinal tract. The histological findings concerning the kidneys had differences in the extent and frequency of changes between the dosage and between male and female rats.

Kidneys:
- Chronic appearance of glomeruli indicating moderate level of atrophy (male/females of 0.6% and 1.8% treated groups)
- Glomeruli contributed to overall swelling through dilation of vessel lumen and interstitial swelling (significant in male/female rats of 0.2 and 0.07% dose groups. They were not very significant in the 1.8 and 0.6% dose groups)
- The glomerular interstitum had thawn mildly but chronically (present in control group but was significant for male/female rats in 1.8 and 0.6% groups)
- Cloudy swelling/vacuolar degeneration of proximal renal tubular epithelial cells were seen (mildly present in control group and slightly significant in LAS treated groups).
- Small, yellowish brown blobs the size of nuclei was observed in the proximal renal tubular epithelial cells (significantly in male/females of 1.8% dose group)
- Blue lead color exhibited on the lumen of the Henle loop through H.E. staining, while substances giving a strong positive PAS response were forming layers in circular forms or were present in small pieces
-Lumen dilation of the paroximal renal tube and flattening of epithelial cells (significantly in males of 0.2% dose group, no effect observed in females and control groups)
- The renal tubule had collapsed (significantly in male/females of 1.8% dose group)
- Swelling of surrounding connective tissue was observed in the interstitium (observed in control group and significantly in males of LAS-0.2% dose group and both male and female rats of 0.6% dose group)

- Various changes to the kidneys as seen in 0.2% to 0.07% dose groups differ in their extent and frequency of incidence, but these changes were also seen in the control group, so they were not considered changes that lead to permanent damages like collapse of renal tubule or cyst formation.

Liver: Disappearance of basophilic substances in the hepatocytes and eosin thickening of hepatocytes were observed mildly in male and female rats of the 1.8% dose group.

Intestinal tract: Decrease in height of epithelial cells in the colonic mucosa and disappearance of vacuoles from goblet cells were observed mildly in female rats of the 1.8 and 0.6% dose groups.

Further details on are provided in "Any other information on results inc.tables"
Key result
Dose descriptor:
NOAEL
Effect level:
40 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: Suppression of weight gain, increase in appendix weight, alteration in hematological and clinical biochemistry parameters, and/ or tissue damages to the large intestines, liver and kidneys in dose levels at ≥ 115 mg/kg bw/day (0.2% diet).
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
115 mg/kg bw/day (nominal)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1:  Body weight gain, food, detergent, water intake and feces excretion in males (Yoneyama et al., 1972)

Group  No. of Rats Initial body wt (g) (mean±S.E) Final body wt (g) (mean±S.E) Body weight gain (1) (%) (mean±S.E) Food (g/rat/day) Detergent water (g/rat/day) Feces (g/rat/day)
mg/rat/day mg/kg/day
Control 10 85±0.871 387±9.509 452.1±9.841 14.17 - - 16.9 2.31
1.80% 10-9 85.2±0.553 290.00±5.206 341.09±5.136** 9.84 177.12 814.8 13.29 1.78
0.60% 10 86.40±1.543 365.80±8.517 426.23±8.893 13.84 83.04 322.3 16.28 2.23
0.20% 10 84.50±0.957 383.40±6.715 453.92±7.603 13.7 27.4 101.5 18.18 2.28
0.07% 10 80.9±1.923 387.10±5.930 480.74±13.113 14.48 10.14 37.2 16.91 2.24

Table 2:  Body weight gain, food, detergent, water intake and feces excretion in females (Yoneyama et al., 1972)

Group  No. of Rats Initial body wt (g) (mean±S.E) Final body wt (g) (mean±S.E) Body weight gain (%) (1) (mean±S.E) Food (g/rat/day) Detergent water (g/rat/day) Feces (g/rat/day)
mg/rat/day mg/kg/day
Control 10 71.80±0.512 202.50±2.325 282.24±4.422 9.35 - - 17.48 1.53
1.80% 10 74.30±0.943 158.80±3.411 214.02±5.288** 7.01 126.18 913 12.14 1.23
0.60% 10 74.10±0.673 195.80±4.149 264.25±5.207* 9.04 54.24 332.2 13.99 1.53
0.20% 10 74.30±1.116 201.10±3.334 271.05±5.314 8.9 17.8 108.7 17.69 1.39
0.07% 10 76.00±2.683 203.20±7.172 269.35±10.255 9.35 6.55 39.4 19.56 1.63

where (1) refers to Final BW/Initial BW-1) into100

*P<0.05

**P<0.01

Table 3 : Hematology-I in males and females rats (Yoneyama et al., 1972)

Group No. of Rats RBC*104/mm3  Ht (%) Hb (g/dl) MCH (rr) MCV (µ8) MCC%
Male
Control 10 910±13 53.1±0.4 19.8±0.3 21.8±0.4 58.4±1.0 37.4±0.6
1.80% 9 924±271) 54.6±0.4 20.6±0.5 22.4±0.8 59.3±1.3 37.7±0.8
0.60% 10 890±14 54.0±0.4 19.8±0.1 22.3±0.3 60.7±0.9 36.7±0.1
0.20% 10 914±25 53.3±0.3 19.5±0.1 21.5±0.6 58.6±1.6 36.7±0.2
0.07% 10 925±20 53.3±0.4 19.6±0.1 21.2±0.5 57.8±1.2 36.7±0.2
Female
Control 10 838±22 52.1±0.5 19.9±0.5 23.9±0.9 62.5±1.6 38.2±1.0
1.80% 10 785±14 49.5±0.3** 18.0±0.1** 23.0±0.4 63.2±0.9 36.4±0.1
0.60% 10 795±17 50.9±0.3 19.0±0.1 24.0±0.5 64.2±1.1 37.4±0.2
0.20% 10 778±16 51.1±0.2 19.1±0.1 24.7±0.4 65.9±1.3 37.5±0.2
0.07% 10 806±18 50.6±0.4 18.9±0.1 23.6±0.6 63.0±1.5 37.4±0.3

Table 4 : Hematology-II in males and females rats (Yoneyama et al., 1972)

Group No. of Rats Thrombocyte*104(A) Osmotic fragility (A) Whole blood clotting time (min)
Beginning Ending
Male
Control 10 70.5±9 (8) 213.6±9.4 (9) 1.06±0.13 2.12±0.26
1.80% 9 70.2±4.9 (8) 199.9±6.6 (9) 1.14±0.11 2.32±0.27
0.60% 10 71.6±8.4 211.4±7.9 1.42±0.23 2.71±0.29
0.20% 10 64.4±5.5(9) 228.1±6.5(9) 1.34±0.19 2.50±0.35
0.07% 10 80.1±13.2 225.1±10.5 1.40±0.19 2.52±0.33
Female
Control 10 52.0±3.8 (9) 240.4±7.5 (8) 1.29±0.29 2.95±0.30
1.80% 10 58.4±4.5 (9) 181.8±6.5(8)* 1.37±0.14 2.57±0.19
0.60% 10 54.8±3.3 (9) 226.0±5.9 (9) 1.24±0.17 2.81±0.22
0.20% 10 59.8±5.9 (9) 245.4±9.3(9) 1.26±0.19 2.92±0.22
0.07% 10 63.3±5.3 (8) 248.4±10.2 1.22±0.21 3.30±0.41

*P<0.05

**P<0.01

(A): No of rats

1) (mean±SE)

Table 5: Percent differential of Leucocytes (Yoneyama et al., 1972)

Group No. of Rats Neutro
Lympho Mono Eosino
Male
Control 10 9.8±0. 88 (1) 89.08±0.954 0.28±0.102 0.83±0.203
1.80% 9 12.09±0.698* 87.22±0.624 0.09±0.061 0.58±0.164
0.60% 10 10.77±1.017 87.82±0.860 0.26±0.094 0.60±0.158
0.20% 10 10.34±0.521 88.22±0.625 0.40±0.102 0.62±0.128
0.07% 10 9.41±1.074 89.35 ± 0.998 0.50± 0.088 0.73±0.125
Female
Control 10 8.24±0.56 90.85±0.685 0.18±0.073 0.72±0.150
1.80% 10 8.67±0.747 90.61 ± 0.762 0.28±0.076 0.42±0.108
0.60% 10 10.21±1 .054 88.49±1.065 0.32±0.071 0.96±0.143
0.20% 10 8.75±0.754 90.15±0.781 0.38±0.096 0.70±0.146
0.07% 10 8.65±0.752 90.39 ± 0.771 0.40±0.099 0.54±0.108

Table 6: Counts differential of leucocytes.102/mm3 (Yoneyama et al., 1972)

Group No. of Rats WBC Neutro Lympho Mono Eosino
Male
Control 10 88.12±3.686 (1) 8.49±0.678 78.65±3.779 0.24±0.087 0.67±0.148
1.80% 9 96.16±5.726 11.78±1.113 83.76±4.805 0.08 ±0.057 0.54±0.142
0.60% 10 91.47±5.430 9.96±1.154 80.27±4.686 0.25±0.088 0.58±0.142
0.20% 10 90.55±6.435 9.61±1.166 79.64±5.196 0.36 ±0.089 0.60±0.155
0.07% 10 96.15±3.329 9.08±1.077 85.85±2.913 0.50±0.093 0.71±0.135
Female
Control 10 92.00±3.802 7.59±0.579 83.57±3.517 0.16±0.066 0.67±0.139
1.80% 10 109.30±3.831** 9.34±0.664 95.03 ± 7.208 0.31 ± 0.085 0.47±0.120
0.60% 10 87.90±3.631 8.87±0.847 77.89±3.679 0.26±0.058 0.85±0.133
0.20% 10 83.15±4.350 7.38±0.637 76.67 ± 5.063 0.32±0.078 0.61 ± 0.138
0.07% 10 94.40±5.110 8.36±1.176 85.13±4.180 0.37±0.091 0.51 ± 0.101

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 7: Serum Chemistry (Yoneyama et al., 1972)

Group No. of Rats ALP
K-A Unit/dl
LAP
G-R Unit
GOT
karmen unit/ml
UN
mg/dl
A/G ratio Total protein
g/dl
Bililubin
                (Indirect)
mg/dl
(Direct)
mg/dl
Total
mg/dl
Male      
Control 10 24.07±1.1091) 185.81± 9.586 132.75± 6.357 16.92±1.102 1.42±0.082 7.36±0.071 0.20±0.050 0.11±0.036 0.29±0.044
1.80% 9 44.96±4.01** 179.80± 9.74 139.12± 9.397 16.30±0.634 1.59±0.129 6.55±0.191** 0.40±0.250 0.12±0.027 0.48±0.268
0.60% 10 29.22±1.093** 180.54± 7.897 133.50± 5.559 15.76±0.611 1.46±0.096 7.20±0.084 0.30±0.201 0.32±0.211 0.56±0.415
0.20% 10 24.34±1.173 181.23± 9.637 140.25±14.732 13.04±1.224* 1.47±0.091 7.20±0.059 0.41±0.231 0.31±0.188 0.68±0.422
0.07% 10 25.48±0.816 178.99± 9.4271 144.60±11.501 15.36±1.221 1.38±0.082 7.26±0.118 0.35±0.201 0.34±0.197 0.64±0.401
Female      
Control 10 24.41±1.380 198.54± 8.899 156.30±12.716 17.52±1.733 1.53±0.062 7.80±0.122 0.12±0.050 0.14±0.054 0.23±0.054
1.80% 10 61.41±6.124** 208.72± 9.097 154.60±8.297 19.00±1.043 1.76±0.121 6.42±0.127** 0.12±0.025 (9) 0.12±0.033 (9) 0.25±0.054 (9)
0.60% 10 31.80±1.599** 187.58± 10.642 139.50±3.024 17.15±2.020 1.70±0.097** 7.22±0.186* 0.14±0.036 0.12±0.038 0.27±0.062
0.20% 10 27.23±1.049 193.65± 7.686 146.55±9.144 14.14±1.404 1.48±0.164 7.38±0.184 0.20±0.044 (9) 0.11±0.023 (9) 0.31±0.052 (9)
0.07% 10 25.77±1.237 192.23± 8.056 128.25±5.432 15.09±1.245 1.72±0.067 7.68±0.163 0.15±0.059 (9) 0.10±0.018 (9) 0.24±0.06 (9)

where:

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 8 : Urinalysis in female and male rats (Yoneyama et al., 1972)

Group No. of Rats pH Protein Glucose Keton Blood Bilirubin Urobilin
    8 7 6 5 - ± + ++ +++ - + - ± + - ± + - + 0.1* 1
Male
Control 10 1 4 5 - - - 7 2 1 10 - 7 3 - 10 - - 10 - 10 -
1.80% 9 - - 9 - - - 2 3 4 9 - 2 7 - 9 - - 9 - 9 -
0.60% 10 - 5 5 - - 2 5 2 1 10 - 5 5 - 10 - - 10 - 10 -
0.20% 10 - 2 8 - - 4 3 3   10 - 6 4 - 10 - - 10 - 10 -
0.07% 10 - 4 6 - 1 - 6 1 2 10 - 8 2 - 10 - - 10 - 10 -
Female
Control 10 2 2 6 - 2 2 5 1 - 10 - 5 5 - 10 - - 10 - 10 -
1.80% 10 1 - 9 - 3 3 4 - - 10 - 4 5 - 10 - - 10 - 9 -
0.60% 10 - 5 5 - 2 3 2 3 - 10 - 7 3 - 10 - - 10 - 10 -
0.20% 10 - 3 7 - 2 2 5 1 - 10 - 4 6 - 10 - - 10 - 10 -
0.07% 10 - 3 7 - 3 3 4 - - 10 - 7 3 - 10 - - 10 - 9 -

*Ehrlich units

Table 9: Organ Weights (Male-1) (Yoneyama et al., 1972)

Group  No. of rats Heart
[Actual weight (mg)/weight (mg)/100 g]
Spleen
[Actual weight (mg)/weight (mg)/100 g]
Thymus
[Actual weight (mg)/weight (mg)/100 g]
Liver
[Actual weight (g)/weight (g)/100 g]
Lung
[Actual weight (mg)/weight (mg)/100 g]
Kidneys
[Actual weight (mg)/weight (mg)/100 g]
Hypophysis
[Actual weight (mg)/weight (mg)/100 g]
  R L
Control 10 930±13/241± 31) 614±14/158±2 129±5.7/33±0.9 12.0±0.38/3.1±0.04 1211±44/314±13 1105±33/285± 6 1126±33/219± 6 10.0±0.73/2.6±0.21
1.80% 9 694±11**/239±3 460±11**/158±2 102±6.3**/35±1.7 10.3±0.28**/3.5±0.08** 971±35**/324±13 836±19**/288±3 852±22**/294±5 8.8±0.42/3.0±0.15
0.60% 10 878±18*/235± 4 572±15*/153± 4 122±8.8*/32± 2.3 12.0±0.31/3.2±0.07 1080±21/289±6 1045±31/280±9 1049±34/281±9 9.4±0.30/2.5±0.08
0.20% 10 918±16/239± 2 644±27/168±7 130±9.0/33±2.1 12.3±0.42/3.2±0.12 1253±63/326±14 1083±34/282±7 1071±28/279±4 8.9±0.48/2.3±0.13
0.07% 10 932±16/240± 2 641±12/165± 2 129±10.8/33± 2.5 12.0±0.31/3.1±0.05 1285±47/332±12 1074±25/277±5 1083±52/279±12 9.3±0.50/2.4±0.13

Table 10: Organ Weights (Male-2) (Yoneyama et al., 1972)

Group  No. of rats Thyroid glands
[Actual weight (mg)/weight (mg)/100 g]
Supranal gland
[Actual weight (mg)/weight (mg)/100 g]
Testis
[Actual weight (mg)/weight (mg)/100 g]
Prostate glands
[Actual weight (mg)/weight (mg)/100 g]
Brain
[Actual weight (mg)/weight (mg)/100 g]
Caecum
[Actual weight (mg)/weight (mg)/100 g]
  R L R L
Control 10 14.4±0.4211)/3.7±0.16 18.0±0.61/4.6±0.18 19.3±0.44/5.0±0.17 1592±20/412± 7 1622±21/420± 6 252±16/65±4.8 1983±12/515±11 837±34/216± 8
1.80% 9 11.5±0.76**/3.9±0.24 15.5±0.44**/5.3±0.17* 17.1±0.65*/5.9±0.27* 1513±23*/522±7** 1501±36**/518±13** 231±21/79±7.0   1960±22/677±11 ** 1484±30**/512±12**
0.60% 10 13.2±0.69/3.5±0.22 17.5±0.76/4.6±0.20 18.2±0.69/4.8±0.19 1576±27/422± 11 1612±36/432±11 231±21/61±5.7 1969±18*/528±12 939±41/251±11*
0.20% 10 13.3±0.57/3.4±0.11 17.3±0.61/4.5±0.17 18.6±0.84/4.8±0.23 1564±22/408± 5 1621±20/423± 4 298±21/78±5.9 2020±11/528±8 971±35*/253 ± 7**
0.07% 10 13.6±0.54/3.5±0.13 16.8±0.85/4.3± 0.22 18.7±0.44/4.8± 0.11 1643±14/425± 6 1676±15/434± 7 318±21*/82±6.1 1918±106/494±25 938±38/242± 9

where,

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 11: Organ Weights (Female-1) (Yoneyama et al., 1972)

Group  No. of rats Heart
[Actual weight (mg)/weight (mg)/100 g]
Spleen
[Actual weight (mg)/weight (mg)/100 g]
Thymus
[Actual weight (mg)/weight (mg)/100 g]
Liver
[Actual weight (g)/weight (g)/100 g]
Lung
[Actual weight (mg)/weight (mg)/100 g]
Kidneys
[Actual weight (mg)/weight (mg)/100 g]
Hypophysis
[Actual weight (mg)/weight (mg)/100 g]
R L
Control 10 596±1001)/265± 6 405± 5/200±4 134±7.7/66±3.9 6.5±0.12/3.2±0.08 822±19/409±11 720±22/ 359±12 738±18/395±11 13.9±0.76/6.8±0.39
1.80% 10 426± 11**/269±7* 294± 10**/185± 6 102±6.8**/64±4.1 6.3±0.24/3.9±0.12** 673±20**/425±16 546±16**/345±13 563±24**/357±21 12.3±1.08/7.7±.63
0.60% 10 559± 11*/286± 5 387 ± 7/198±5 125±5.8/63±2.2 6.5±0.22/3.3±0.10 793±14/406±8 664±16/340± 9 715±32/ 365±14 16.2±0.94/8.3±0.46*
0.20% 10 574±13/258± 5 405 ±8/201 ± 4 131±4.6/65±2.6 6.5±0.18/3.2±0.07 802±26/398±10 654 ±20*/325± 7 665±28*/330±11* 15.0±1.10/7.4±0.53
0.07% 10 596±16/294± 3 408±13/201± 4 132±2.8/65±2.3 6.7±0.30/3.3±0.05 801±19/396±11 717±23/355±13 738±33/364±13 16.3±0.94/8.0±0.40

Table 12: Organ Weights (Female-2) (Yoneyama et al., 1972)

Group  No. of rats Thyroid glands
[Actual weight (mg)/weight (mg)/100 g]
Supranal gland
[Actual weight (mg)/weight (mg)/100 g]
Ovary
[Actual weight (mg)/weight (mg)/100 g]
Uterus
[Actual weight (mg)/weight (mg)/100 g]
Brain
[Actual weight (mg)/weight (mg)/100 g]
Caecum
[Actual weight (mg)/weight (mg)/100 g]
  R L R L
Control 10 11.2±0.3211)/ 5.5±0.17 21.9±0.99/10.8±0.64 23.2±1.24/11.4±0.94 23.3±1.0/11.5±0.5 24.4±1.1/12.0±0.5 638±27/315±13 1832±16/906±14 681±18/337±11
1.80% 10 10.9±0.73/6.9±0.53* 15.6±0.81**/9.8±0.52 16.8±0.84**/10.6±0.55 17.6±1.4**/11.1±0.9 18.2±1.8**/11.5±1.1 292±18**/185±12** 1780±15*/1126±28 929 ± 26**/586±14**
0.60% 10 13.9±1.30/7.1±0.70 22.9±0.56/11.7±0.36 24.0±0.61/12.3±0.44 22.8±1.2/11.7±0.7 24.5±1.0/12.6±0.6 618±32/315±15 1840±14/943±20 697±21/356± 7
0.20% 10 12.2±0.96/6.0±0.46 21.7±1.11/10.7±0.49 22.2±0.92/11.0±0.43 23.6±1.0/11.7±0.4 23.3±1.2/11.5±0.5 594±31/296±17 1837±18/915±13 688±26/343±14
0.07% 10 12.9±0.93/6.4±0.55 22.6±0.95/11.1±0.35 24.1±0.98/11.8±0.33 23.0±1.9/11.2±0.6 24.6±1.5/12.0±0.5 592±18/292± 8 1827±18/908±30 753±25*/373±13

where,

*P<0.05

**P<0.01

(A): No of rats

1)(mean±SE)

Table 13: Summary of microscopic  changes in kindey (Yoneyama et al., 1972)

Microscopic findings # Male Female
C LAS C LAS
0.07% 0.2% 0.6% 1.8% 0.07% 0.2% 0.6% 1.8%
Swelling in glomerulus S E
M O
S L
0
2 (10)
2
0
5 (10)
4
1
6 (10)
3
0
7 (10)
2
0
5 (9)
4
0
2 (10)
3
1
5 (10)
4
1
5 (10)
4
0
7 (10)
3
0
5 (10)
5
Histolysis in glomerulus S E
M O
S L
0
2
2
0
7
2
1
5
4
1
5
4
1
5
3
0
3
1
0
5
5
0
9
4
1
6
3
1
5
4
Atrophy in glomerulus S E
M O
S L
0
1
1
0
5
5
1
6
2
2
7
1
3
6
0
0
1
1
0
4
5
0
6
3
1
7
2
4
5
1
Cloudy swelling in epithelium of proximal tubules S E
M O
S L
0
0
4
0
1
5
0
6
2
0
8
2
0
7
2
0
0
3
0
0
4
0
3
3
0
3
3
0
4
5
Vacuolation in epithelium of proximal tubules S E
M O
S L
0
0
4
0
6
4
0
3
7
0
4
6
0
5
4
0
1
5
0
7
3
0
6
4
0
7
3
0
7
3
Lipofuscin in epithelium of proximal tubules S E
M O
S L
0
0
0
0
0
0
0
0
5
0
0
5
2
7
0
0
0
0
0
0
0
0
0
1
0
0
4
5
5
0
PAS-positive materies in loop of Henle S E
M O
S L
          0
0
4
0
3
7
0
3
6
0
3
6
0
0
3
Dilatation in lumina of distal tubules S E
M O
S L
0
0
2
0
4
6
3
3
4
2
4
4
0
5
4
0
0
0
0
0
5
0
3
3
0
5
5
0
3
5
Decrease in cell height of epithelium in distal tubules S E
M O
S L
0
0
1
0
0
6
2
3
5
2
3
4
0
4
5
0
0
0
0
0
3
0
0
5
0
3
5
0
3
5
Destruction in urinary tubules S E
M O
S L
0
0
0
0
0
0
0
0
2
0
2
3
0
3
4
0
0
0
0
0
2
0
0
3
0
0
5
0
4
5
edema in stroma S E
M O
S L
0
1
2
1
5
4
1
6
3
1
3
5
0
4
5
0
1
1
1
4
5
1
5
4
1
6
3
0
2
7
Swelling in perivascular connective tissues S E
M O
S L
0
1
1
1
4
4
1
3
6
0
5
5
0
3
4
0
1
1
0
4
5
0
5
5
0
7
3
0
1
6
Proliferation in interstitial connective tissues S E
M O
S L
0
0
1
0
0
5
0
0
5
0
1
4
0
2
6
0
0
2
0
0
2
0
0
5
0
0
6
0
0
6

where, SE: Severe

SL: Slight

MO: Moderate

(A) No. of rats

Table 14:  Summary of microscopic changes in liver and large intestine (Yoneyama et al., 1972)

Microscopic findings # Male Female
C LAS C LAS
0.07% 0.2% 0.6% 1.8% 0.07% 0.2% 0.6% 1.8%
Decrease in basoplam in liver cells S E
M O
S L
0
0 (10)
0
0
0 (10)
0
0
0 (10)
1
0
0 (10)
1
0
0 (9)
6
0
0 (10)
0
0
0 (10)
0
0
0 (10)
0
0
0 (10)
3
0
1 (10)
6
Increase in eosinophilia in liver cells S E
M O
S L
0
0
1
0
0
0
0
0
0
0
0
0
0
0
6
0
0
1
0
0
0
0
0
0
0
0
2
0
2
7
Swelling in liver cells S E
M O
S L
0
0
0
0
0
0
0
0
0
0
3
3
0
3
3
0
0
0
0
0
0
0
1
3
0
1
4
0
2
6
Rarefaction in cytoplasm of liver cells S E
M O
S L
0
0
3
0
0
8
0
1
6
0
1
6
0
1
6
0
0
2
0
0
6
0
1
7
0
1
7
0
1
7
Decrease in cell height of epithelium in large instestine S E
M O
S L
0
0
0
0
0
0
0
0
2
0
0
4
0
0
5
0
0
0
0
0
0
0
0
3
0
1
4
0
1
4

where, SE: Severe

SL: Slight

MO: Moderate

(A) No. of rats

Conclusions:
Administration of linear alkylbenzene sulfonic acid sodium salt (LAS) to Wistar SLC rats at dose levels of 0, 0.07, 0.2, 0.6 and 1.8% (equivalent to 0, 40, 115, 340, 1030 mg/kg bw/day) for 26 weeks revealed an NOAEL of 0.07% (40 mg/kg bw/day), based on tissue damage in caecum, kidney and liver, hematological and/or clinical chemistry parameters changes at higher dose levels (115 mg/kg bw/day and above).
Executive summary:

A sub-chronic repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in Wistar SLC rats. The test substance was administered to groups of 10 male and female rats/dose in the diet for 26 weeks at doses of 0, 40, 115, 340 and 1030 mg/kg/day. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, haematological, clinical chemistry and urine parameters were analysed. Gross pathological examinations were observed after necropsy and the major organs were subjected to histopathological examinations. Significant diarrhoea, suppressed growth, increased caecal weight and degeneration of renal tubes were observed in the highest dose group. Similar but less severe signs were seen in other doses with the exception of the lowest dose of 40 mg/kg/day, which showed no adverse effects related to exposure to the LAS. Under the study conditions, systemic toxicity NOAEL was determined to be 40 mg/kg/day, based on tissue damage in caecum, kidney and liver, haematological as well as clinical chemistry parameters changes at higher dose levels (Yoneyama, 1972). 

Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
A justification of the read-across approach is appended in Section 13.
Reason / purpose:
reference to same study
Reason / purpose:
read-across source
Principles of method if other than guideline:
The purpose of this study is to determine the sub chronic toxicity of LAS in Wistar JCL rats, focusing on the liver and kidneys. Male and female rats were maintained on either test diets (0, 0.6 and 1.8%) or drinking water (0, 0.07 and 0.2%) for 9 months. 0.6 and 1.8% rats in drinking water study exhibited severe weight loss so LAS administration was stopped after 2 weeks. Mortality, clinical observations and body weights were recorded during the study. All the surviving animals were humanely euthanized at end of 9 months and gross necropsy, hematological, serum biochemical tests, enzyme tests on the liver and kidneys were performed and organs weights were measured. No histopathology was performed.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: Wistar JCL
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female rats were obtained from Shizuoka Agricultural Cooperative Association for
Laboratory Animals
- Age at study initiation: 4-weeks old
- Weight at study initiation: 100 - 124 g (male rats), 82 - 100 g (female rats)
- Fasting period before study: No
- Housing: 5 animals were housed per cage.
- Diet: CE-2 food (from CLEA Japan); ad libitum
- Water: ad libitum
- Acclimation period: 1 week before the start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature: 25 ± 1°C
- Humidity: 50 - 60%
- Air changes: Not reported
- Photoperiod: 12 hours dark /12 hours light

IN-LIFE DATES: Not reported
Route of administration:
other: oral: drinking water and feed
Details on route of administration:
LAS was administered to animals by mixing 0.6 and 1.8% in CE-2 food (CLEA Japan) and dissolving to 0.07 and 0.2% in drinking water.
Vehicle:
other: Test substance was administered either in diet or drinking water
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
9 months
Frequency of treatment:
Continuous in diet or drinking water (ad libitum)
Dose / conc.:
0.6 other: %
Remarks:
in diet
Dose / conc.:
1.8 other: %
Remarks:
in diet
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 0.6% dose level)
Dose / conc.:
900 mg/kg bw/day (nominal)
Remarks:
in diet (corresponding to 1.8% dose level)
Dose / conc.:
0.07 other: %
Remarks:
in drinking water
Dose / conc.:
0.2 other: %
Remarks:
in drinking water; 0.6 and 1.8% dose group (equivalent to 857.14 and 2571.43 mg/kg bw/day) were also included, however due to severe weight loss so LAS administration was stopped after 2 weeks.
No. of animals per sex per dose:
Feeding study (mixed in diet): 8 animals/sex/dose
Drinking water study: 9 animals/sex/dose
Control animals:
yes, concurrent vehicle
yes, plain diet
Positive control:
No
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption: Yes
- Compound intake: Yes
- Time schedule for examinations: Weekly

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: Weekly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 9 months, blood was collected from each animal after euthanizing.
- Anesthetic used for blood collection: No
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters: White blood cells (WBC), red blood cells (RBC), Hemoglobin (Hgb), Hematocrit (Hct), mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 9 months, blood was collected from each animal after euthanizing.
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters: Glutamate oxaloacetate transaminase (GOT), glutamate pyruvic transaminase (GPT), glucose content, urea nitrogen, total cholesterol, albumin, alkaline phosphatase (ALP) and cholinesterase

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER:
- Liver enzyme tests: Glucose 6-phosphatase (G6Pase), lactase dehydrogenase (LDH) and G6P-DH activity
- Kidney enzyme tests: G6Pase, LDH, GPT, GOT, ALP, acid phosphatase (ACP), Na, K-ATPase, and malate dehydrogenase (MDH)
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (However, no details in study report in mentioned)
Gross findings were obtained after euthanizing animals and organs were removed for organ weight measurements.

ORGAN WEIGHTS: Brain, heart, lungs, liver, spleen, kidneys, adrenal gland, testes, ovary, uterus, and appendix
HISTOPATHOLOGY: No
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Both female and male rats (drinking water study) exhibited a slight redness at the tips of their facial fur, and coarse fur all over their bodies.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was significant decrease in body weight gain in males and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
A significant reduction in WBC was observed in 0.6% (diet) male rats and in MCV and MCH was observed in 1.8% (diet) female rats compared to controls.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
There were significant alteration in cholesterol [decrease; all doses, except female rats of 0.07% dose group (drinking water)], GPT [0.6% dose group (diet) females)], GOT [1.8% dose group (diet) males], albumin [1.8% (diet) males] , ALP levels [male and female rats fed with 1.8% LAS-diet] and cholinesterase levels [in male rats fed with 1.8% LAS-diet].
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In males of 1.8% (diet), the absolute liver and testes weight was significantly decreased and the relative testes and liver weight was significantly increased and both absolute and relative spleen weight were also significantly reduced.
In females of 1.8% (diet), absolute and relative liver and caecum weight was significantly increased.
There was also significant decrease in absolute and relative heart weight in females of 0.6% (diet) and 0.07% (drinking water) dose groups.
Gross pathological findings:
not specified
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Liver enzyme tests:
Dietary study: G6Pase activity was reduced in 1.8% dose group males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group males and females, LDH activity was reduced in 0.6, 1.8% dose group. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group and GOT activity was reduced in 1.8% dose group animals.
Drinking water study: LDH activity was reduced in 0.2% dose group males. GOT and GPT activities were clearly reduced in males and GOT activity was increased in 0.07 and 0.2% dose group animals.

Renal enzyme tests:
Dietary study: A significant difference was also observed in G6Pase activity. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals
Drinking water study: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group. A significant difference was also observed in G6Pase activity.
Details on results:
CLINICAL SIGNS: Both female and male rats consuming LAS-containing water exhibited a slight redness at the tips of their facial fur, and coarse fur all over their bodies.

BODY WEIGHT AND WEIGHT CHANGES: There was significant decrease in body weight gain in male and females of 1.8% dose group fed with diet and in males of 0.2% dose group consuming LAS contained water.

WATER CONSUMPTION AND COMPOUND INTAKE: Increased water consumption was observed in males of 1.8% dose group fed with diet and 0.2% dose group consuming LAS contained water.

HAEMATOLOGICAL FINDINGS: A significant reduction in WBC was observed in 0.6% (diet) male rats compared to controls, and a significant reduction in MCV and MCH was observed in 1.8% (diet) female rats compared to controls.

CLINICAL BIOCHEMISTRY: Except female rats of 0.07% dose group (drinking water), a significant reduction or a reduction in cholesterol was observed in male and female rats of all dose groups compared to controls. GPT was significantly reduced in 0.6% dose group (diet) females, and reduced in females of other treatment groups. GOT was significantly reduced in 1.8% dose group (diet) males, and reduced in both females and males of other treatment groups. Albumin was significantly reduced in 1.8% (diet) males and reduced in males of 0.2% dose group (drinking water) and females of 1.8% dose group (diet). There was significant increase in ALP levels in male and female rats fed with 1.8% LAS-diet and cholinesterase levels in male rats fed with 1.8% LAS-diet.

ORGAN WEIGHT: In males of 1.8% (diet), the absolute liver and testes weight was significantly decreased and the relative testes and liver weight was significantly increased and both absolute and relative spleen weight were also significantly reduced. In females of 1.8% (diet), absolute and relative liver and caecum weight was significantly increased. There was also significant decrease in absolute and relative heart weight in females of 0.6% (diet) and 0.07% (drinking water) dose groups.

LIVER ENZYME TESTS: G6Pase activity was reduced in 1.8% dose group (diet) males and females, G6P-DH activity was reduced in 0.6 and 1.8% dose group (diet) males and females, where the percentage reduction was greater in 1.8% dose group (diet) animals. LDH activity was clearly reduced in 0.6, 1.8% dose group (diet), and 0.2% dose group (drinking water) males, but reduced in only 1.8% dose group (diet) females. GOT and GPT activities were clearly reduced in males, while in females GPT activity was reduced only in 1.8% dose group (diet) animals, and GOT activity was increased in 0.07 and 0.2% dose group (drinking water) animals but reduced in 1.8% dose group (diet) animals.

RENAL ENZYME TESTS: In males, Na, K-ATPase activity was significantly reduced in 0.2% dose group (drinking water) animals, and also reduced in other male treatment groups. A significant difference was also observed in G6Pase activity, where the reduction observed was associated with an increase in amount consumed. In females, G6Pase, Na, K-ATPase, and LDH activity were significantly reduced in 1.8% dose group (diet) animals, and G6Pase and LDH activity were also reduced in other treatment groups.
Key result
Dose descriptor:
LOAEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
act. ingr.
Remarks:
(dietery study)
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
water consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
other: Liver and kidney enzyme levels
Remarks on result:
other: Adverse effects were observed at all dose levels
Key result
Dose descriptor:
NOAEL
Effect level:
85 mg/kg bw/day (nominal)
Based on:
act. ingr.
Remarks:
(drinking water study)
Sex:
male/female
Basis for effect level:
clinical biochemistry
other: Liver and kidney enzyme levels
Remarks on result:
other: Based on significant decreases in the activities of glutamate-oxalate transaminase and lactate dehydrogenase in males and significant decrease in renal Na,K-ATPase in males and females at 145 mg/kg bw/day (0.2% drinking water).
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
145 mg/kg bw/day (nominal)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
145 mg/kg bw/day (nominal)
System:
hepatobiliary
Organ:
liver
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Body weight gain, food, water and sample consumption of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  No. of rats  Initial body weight (g) Final Body weight (g) Body weight gain (%) Food (g/rat/day) Water (g/rat/day) Sample (g/kg BW/day)
Male Control 8 113.1± 1.156 421.6 ± 8.542 372.6 ± 5.889 15 23 -
0.6 F 8 110.6 ± 0.925 413.5 ± 7.356 373.8 ± 6.380 16 23 0.234
1.8 F 8 109.2 ± 2.218 341.1 ± 8.008  312.5 ± 6.518** 14 33 0.747
0.07 W  9 110.6 ± 1.572 411.0 ± 5.255 371.9 ± 6.908  17 29 0.051
0.2 W 9 108.3 ± 1.900 385.1 ± 5.453 356.1 ± 6.346 ** 15 33 0.148
Female Control 8 92.3 ± 1.592 212.6 ± 4.508 230.5 ± 5.420 10 18 -
0.6 F 8 92.1 ± 1.663 216.1 ± 7.705 234.3 ± 5.753 10 17 0.287
1.8 F 8 90.6 ± 1.463 184.8 ± 4.278 204.3 ± 5.791 ** 9 18 0.969
0.07 W  9 92.7 ± 1.770 206.0 ± 3.555 222.4 ± 4.478 11 20 0.082
0.2 W 9 91.6 ± 1.893 209.3 ± 5.383 228.4 ± 3.857 12 18 0.173

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 2: Blood components of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  WBC ( x 103) RBC ( x 106) HGB (g) HCT (%) MCV (µ3) MCH (µµg) MCHC (%)
Male Control 5.5±0.414 5.7 ± 0.142 10.9 ± 0.195 28.0 ± 0.663 49.0 ± 0.189 19.5 ± 0.247 39.5 ± 0.504
0.6 F 4.2 ± 0.221 * 5.9 ± 0.159 11.2 ± 0.291 29.2 ± 1.059 49.6 ± 0.778 19.4 ± 0.436 38.9 ± 0.851
1.8 F 4.9 ± 0.461 5.8 ± 0.076 10.7 ± 0.118 27.8 ± 0.561 49.2 ± 0.250  19.1 ± 0.268 38.5 ± 0.499
0.07 W  4.7 ± 0.209 5.9 ± 0.072 11.3 ± 0.094 29.1 ± 0.317 49.2 ± 0.147 19.1 ± 0.210  38.6 ± 0.447
0.2 W 4.9 ± 0.304 6.0 ± 0.119 11.3 ± 0.141 29.3 ± 0.566 48.8 ± 0.200 19.1 ± 0.238 38.7 ± 0.493
Female Control 2.6 ± 0.297 5.6 ± 0.163 11.0 ± 0.243 28.8 ± 0.859 51.7 ± 0.164 20.0 ± 0.364  38.4 ± 0.689
0.6 F 2.5 ±0.150 5.7 ± 0.265 11.2 ± 0.485 29.3 ± 1.400 51.2 ± 0.313 19.8 ± 0.356 38.5 ± 0.699
1.8 F 3.7 ± 0.556 5.6 ± 0.294 10.5 ± 0.599 28.2 ± 1.546 50.0 ± 0.378 ** 18.9 ± 0.193* 37.5 ± 0.294
0.07 W  2.4 ± 0.213 6.0 ± 0.302 11.5 ± 0.556 30.5 ± 1.551 51.5 ± 0.176  19.5 ± 0.159 37.7 ± 0.299
0.2 W 2.7 ± 0.328 5.6 ± 0.100 11.0 ± 0.176 28.9 ± 0.624 52.0 ± 0.463 20.0 ± 0.349 38.3 ± 0.601

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 3: Biochemical values in serum of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  GOT (K.U/mL) GPT (K.U/mL) GLU (mg/dL) UN (mg/dL) CHO (mg/dL) ALB (g/dL) ALP ((KA-U/dL) ChE (ΔpH)
Male Control 151.3 ± 7.285   107.5 ± 8.177 163.0 ± 11.488 21.0 ± 0.333 77.5 ± 3.417  4.87 ± 0.055 41.3 ± 1.308 0.166 ± 0.010
0.6 F 133.2 ± 5.573 102.3 ± 8.485 132.2 ± 4.139* 21.0 ± 0.684 71.6 ± 2.314 47.2 ± 0.075 44.7 ± 2.782  0.187 ± 0.013
1.8 F 126.7 ± 6.439* 93.0 ± 3.665 144.7 ± 10.099 23.1 ± 0.490** 53.0 ± 1.282** 44.7 ± 0.068** 82.3 ± 3.731** 0.281 ± 0.007 **
0.07 W  142.8 ± 10.111 108.5 ± 12.381 131.6 ± 3.088* 22.8 ± 0.647* 68.2 ± 2.527* 47.5 ± 0.059 44.0 ± 1.239 0.131 ± 0.015
0.2 W 143.8 ± 9.041 103.3 ± 9.513 165.1 ± 14.043 21.0 ± 1.462 62.5 ± 2.534** 43.2 ± 0.251 34.4 ± 3.193 0.171 ± 0.008
Female Control 142.3 ± 14.359 104.2 ± 12.474 135.5 ± 5.246 23.2 ± 0.793 95.8 ± 4.778 47.1 ± 0.265 45.0 ± 5.728 0.963 ± 0.073
0.6 F 116.1 ± 5.244 68.7 ± 2.670* 126.4 ± 3.657 21.5 ± 0.602 86.7 ± 4.230 4.85 ± 0.095 40.7 ± 1.539 1.056 ± 0.023
1.8 F 133.6 ± 4.007 88.1 ± 4.362 130.8 ± 3.945 26.7 ± 1.935 67.8 ± 7.472** 42.8 ± 0.097 73.8 ± 11.808* 0.861 ± 0.053
0.07 W  121.2 ± 7.003 74.2 ± 13.529 127.2 ± 2.035 21.9 ± 0.872 95.0 ± 3.755 49.0 ± 0.187 45.6 ± 12.960 0.881 ± 0.171
0.2 W 128.5 ± 12.209 85.7 ± 9.362 136.1 ± 10.070 21.7 ± 0.950 77.2 ± 8.826 46.8 ± 0.155 44.0 ± 5.756 1.053 ± 0.042

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 4:  Organ weight of male and female rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  Brain  Heart Lung Liver Spleen Kidney (R) Kidney (L) Adrenal (R) Adrenal (L) Testis(R) Testis(L) Ovary (R) Ovary (L) Caecum Uterus
g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g g g/100g
Male Control 1.987 ± 0.011 0.463 ± 0.008 1.026 ± 0.018 0.239 ± 0.005 1.263 ± 0.033 0.294 ± 0.008 12.7 ± 0.341 2.9 ± 0.039 0.698 ± 0.014 0.162 ± 0.003 1.162 ± 0.40 0.270 ± 0.008 1.184 ± 0.041 0.275 ± 0.007 18.3 ± 1.117 4.2 ± 0.217 18.1 ± 0.934  4.2 ± 0.225 1.591 ± 0.023 0.370 ± 0.006 1.643 ± 0.027 0.382 ± 0.005 - - - - 1.238 ± 0.068 0.281 ± 0.015 - -
0.6 F 1.982 ± 0.011 0.473 ± 0.007 0.971 ± 0.022* 0.231 ± 0.004 1.269 ± 0.042 0.302 ± 0.005 12.9 ± 0.310 3.0 ± 0.049 0.682 ± 0.019 0.162 ± 0.002 1.210 ± 0.026 0.290 ± 0.006 1.214 ± 0.032 0.289 ± 0.007 19.6 ± 1.900 4.7 ± 0.435 17.6 ± 1.602 4.2 ± 0.416 1.563 ± 0.019 0.373 ± 0.004 1.585 ± 0.027 0.378 ± 0.007 - - - - 1.303 ± 0.057 0.305 ± 0.010 - -
1.8 F 1.827 ± 0.131 0.521 ± 0.037 0.825 ± 0.026* 0.235 ± 0.003 1.040 ± 0.025** 0.295 ± 0.002 11.5 ± 0.352* 3.2 ± 0.043** 0.501 ± 0.163** 0.143 ± 0.003** 0.977 ± 0.022** 0.279 ± 0.007 0.982 ± 0.017** 0.281 ± 0.007 17.5 ± 1.370 5.0 ± 0.481 17.0 ± 0.654 4.8 ± 0.146 1.455 ± 0.027** 0.415 ± 0.004** 1.491 ± 0.033** 0.425 ± 0.004** - - - - 2.205 ± 0.141 ** 0.647 ± 0.032** - -
0.07 W  1.995 ± 0.016 0.471 ± 0.006 0.981 ± 0.031 0.232 ± 0.008 1.287 ± 0.021 0.304 ± 0.007 13.2 ± 0.258 3.0 ± 0.052 0.682 ± 0.009 0.161 ± 0.002 1.221 ± 0.018 0.288 ± 0.005 1.254 ± 0.012 0.296 ± 0.005* 18.6 ± 0.680  4.4 ± 0.156 19.2 ± 1.024 4.5 ± 0.262 1.606 ± 0.022 0.379 ± 0.006 1.566 ± 0.056 0.370 ± 0.015 - - - - 1.338 ± 0.040 0.314 ± 0.009 - -
0.2 W 1.881 ± 0.108 0.478 ± 0.028 0.975 ± 0.183 0.247 ± 0.003 1.154 ± 0.037* 0.293 ± 0.007 11.6 ± 0.184 2.9 ± 0.042 0.595 ± 0.013** 0.150 ± 0.002 1.124 ± 0.025 0.286 ± 0.007 1.154 ± 0.031 0.293 ± 0.008 18.4 ± 0.914  4.6 ± 0.232 17.5 ± 1.309 4.4 ± 0.354 1.483 ± 0.059 0.377 ± 0.016 1.589 ± 0.013 0.404 ± 0.005 - - - - 1.210 ± 0.035 0.309 ± 0.008 - -
Female Control 1.853 ± 0.023 0.846 ± 0.016 0.647 ± 0.013 0.294 ± 0.003 0.870 ± 0.026 0.397 ± 0.012 6.2 ± 0.150 2.8 ± 0.051 0.441 ± 0.016 0.201 ± 0.006 0.650 ± 0.021 0.296 ± 0.006 0.650 ± 0.021 0.296 ± 0.008 24.8 ± 1.381  11.2 ± 0.443 25.7 ± 1.385 11.7 ± 0.487 - - - - 29.3 ± 2.738 13.4 ± 1.226 31.2 ± 4.007 14.2 ± 1.857 0.828 ± 0.010 0.377 ± 0.005 0.687 ± 0.037 0.313 ± 0.017
0.6 F 1.859 ± 0.011 0.845 ± 0.027 0.595 ± 0.019* 0.269 ± 0.006** 0.810 ± 0.032 0.367 ± 0.015 6.6 ± 0.355 2.9 ± 0.061 0.425 ± 0.010 0.193 ± 0.009 0.648 ± 0.023 0.292 ± 0.004 0.655 ± 0.026 0.296 ± 0.008 25.8 ± 1.563 11.7 ± 0.820 24.8 ± 1.641 11.3 ± 0.863 - - - - 23.3 ± 3.504 10.8 ± 1.740 32.8 ± 3.120 14.9 ± 1.380 0.859 ± 0.064  0.381 ± 0.015 0.660 ± 0.033 0.301 ± 0.020
1.8 F 1.788 ± 0.031 0.954 ± 0.015** 0.455 ± 0.008 0.242 ± 0.002 0.761 ± 0.028 0.405 ± 0.009 7.1 ± 0.195** 3.8 ± 0.082** 0.351 ± 0.012 ** 0.187 ± 0.004 0.580 ± 0.015 0.309 ± 0.003 0.586 ± 0.021* 0.312 ± 0.007 19.6 ± 1.981* 10.3 ± 0.924 21.3 ± 1.487* 11.3 ± 0.609 - - - - 25.3 ± 2.853 13.4 ± 1.417 31.5 ± 2.322 16.7 ± 1.165 1.436 ± 0.080** 0.775 ± 0.039** 0.588 ± 0.066 0.312 ± 0.034
0.07 W  1.854 ± 0.019 0.844 ± 0.016 0.611 ± 0.011* 0.277 ± 0.004** 0.801 ± 0.027 0.365 ± 0.014 6.3 ± 0.139 2.8 ± 0.063 0.424 ± 0.007 0.193 ± 0.004 0.669 ± 0.017 0.304 ± 0.006 0.677 ± 0.010 0.308 ± 0.005 22.0 ± 1.619 10.0 ± 0.720 21.0 ± 1.763 9.8 ± 0.790 - - - - 30.0 ± 1.490 13.6 ± 0.780 31.7 ± 2.300 12.2 ± 1.200 0.785 ± 0.051 0.358 ± 0.024 0.729 ± 0.051 0.331 ± 0.022
0.2 W 1.691 ± 0.112 0.810 ± 0.053 0.593 ± 0.015 0.287 ± 0.004 0.817 ± 0.040 0.393 ± 0.021 6.1 ± 0.220 2.9 ± 0.071 0.428 ± 0.014 0.205 ± 0.006 0.651 ± 0.015 0.312 ± 0.006 0.670 ± 0.020 0.321 ± 0.008* 23.2 ± 1.495 11.1 ± 0.677 24.3 ± 1.154 11.6 ± 0.431 - - - - 28.4 ± 1.633 13.6 ± 0.821 28.6 ± 1.900 13.7 ± 0.830 0.800 ± 0.045 0.379 ± 0.026 0.644 ± 0.032 0.308 ± 0.014

Values: mean ± S.E.

*p<0.05

**p<0.01

Table 5: Liver enzyme activities of rats on administration of LAS for 9 months (Yoneyama et. al., 1976)

Sex  Groups  G6 Pase (µmoles Pi/mg protein /hr G6P-DH (x 10-3)ΔE/mg protein / min LDH ΔE/mg protein / min GPT (x 102) KU/mg protein  GOT (x 102) KU/mg protein 

Male

Control 3.77 ± 0.22 60.3 ± 9.9  11.33 ± 0.53 4.06 ± 0.345 5.80 ± 0.525
0.6 F 3.82 ± 0.50 40.9 ± 9.1 ** 10.28 ± 0.84* 3.94 ± 0.484 5.49 ± 0.810
1.8 F 2.60 ± 0.015** 36.0 ± 7.7** 9.26 ± 0.99** 3.09 ± 0.225** 4.95 ± 0.510**
0.07 W  3.57 ± 0.36 68.0 ± 15.1 10.92 ± 0.91 3.89 ± 0.360 5.61 ± 0.410
0.2 W 3.71 ± 0.35 56.8 ± 14.9 9.28 ± 0.71** 4.10 ± 0.420 6.16 ± 0.545
Female Control 2.73 ± 0.57  143.6 ± 30.0 70.6 ± 11.6 2.71 ± 0.405 4.91 ± 0.635
0.6 F 2.67 ± 0.51 100.7 ± 39.9* 63.4 ± 14.2 2.60 ± 0.505 4.30 ± 0.560
1.8 F 2.17 ± 0.15* 58.5 ± 12.2* 62.2 ± 4.7 2.37 ± 0.225 4.05 ± 0.450 **
0.07 W  2.61 ± 0.20 138.3 ± 41.0 65.8 ± 8.5 2.57 ± 2.290 5.61 ± 0.410*
0.2 W 2.76 ± 0.28 120.9 ± 27.6 66.5 ± 11.3 2.89 ± 0.195

6.16 ± 0.545**

Values: mean ± S.D.

*p<0.05

**p<0.01

Table 6: Kidney enzyme activities of rats on administration of LAS for 9 months  (Yoneyama et. al., 1976)

Sex  Groups  ALP (µmoles/mg/hr) ACP (µmoles/mg/hr) G6Pase (µmoles/mg/hr) Na, K-A Tpase (µmoles/mg/hr) GPT (KU/mL) GOT (KU/mL) LDH (ΔE/mg/min) MDH (ΔE/mg/min)
Male Control 16.37 ± 1.984 1.80 ± 0.188 3.97 ± 0.590 3.94 ± 0.613 23.99 ± 5.300 192.11 ± 35.741 0.339 ± 0.043 2.308 ± 0.0447
0.6 F 14.44 ± 2.046 1.72 ± 0.141 3.92 ± 0.618 3.63 ± 0.640 23.16 ± 5.885 199.95 ± 22.869 0.334 ± 0.050 2.325 ± 0.575
1.8 F 15.74 ± 1.933 1.78 ± 0.179 3.47 ± 0.732 3.32 ± 1.311 21.11 ± 4.690 194.18 ± 13.668 0.358 ± 0.070 2.354 ± 0.554
0.07 W  14.73 ± 2.387 1.85 ± 0.194 4.26 ± 0.547 3.52 ± 1.170  23.85 ± 4.640 207.39 ± 21.308 0.347 ± 0.098 2.617 ± 0.628
0.2 W 15.03 ± 1.169 1.66 ± 0.109 4.03 ± 0.428 3.09 ± 0.224* 24.44 ± 3.535 202.94 ± 15.708 0.362 ± 0.058 2.679 ± 0.740 
Female Control 15.94 ± 3.369 2.02 ± 0.182 3.96 ± 0.545 4.29 ± 1.027 28.27 ± 4.407 188.33 ± 21.189 0.350 ± 0.035 2.387 ± 0.601
0.6 F 17.11 ± 2.919 2.03 ± 0.179 3.63 ± 0.521 4.29 ± 0.656 30.26 ± 5.736 196.48 ± 21.273 0.338 ± 0.044 2.231 ± 0.433
1.8 F 16.86 ± 2.369 1.85 ± 0.195 3.13 ± 0.426** 3.19 ± 0.613* 32.60 ± 3.737 184.89 ± 11.276 0.291 ± 0.025** 2.181 ± 0.404
0.07 W  16.24 ± 2.381 1.98 ± 0.218 3.72 ± 0.443 4.14 ± 1.452 27.19 ± 3.319 198.09 ± 29.803 0.322 ± 0.035 2.052 ± 0.306
0.2 W 15.02 ± 3.244 1.90 ± 0.136 3.45 ± 0.651 4.59 ± 1.154 27.03 ± 4.430 187.36 ± 20.235 0.316 ± 0.052 2.070 ± 0.362 

Values: mean ± S.D.

*p<0.05

**p<0.01

Conclusions:
Administration of LAS to Wistar JCL rats by test diets at dose levels of 0, 0.6 and 1.8% for 9 months (focusing on the liver and kidneys) revealed an LOAEL of 0.6% (300 mg/kg bw/day in diet), based on adverse effects at all dose levels.
Administration of LAS to Wistar JCL rats in drinking water at dose levels of 0, 0.07 and 0.2% for 9 months (focusing on the liver and kidneys) revealed an NOAEL of 0.07% (85 mg/kg bw/day in drinking water), based on adverse effects at all dose levels, based on significant decreases in the activities of glutamate-oxalate transaminase and lactate dehydrogenase in males and significant decrease in renal Na,K-ATPase in males and females at 145 mg/kg bw/day (0.2% drinking water).
Executive summary:

A sub-chronic (9 months) repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in male and female Wistar JCL rats. The test substance was administered at the following dose levels: through diet at 0, 0.6 and 1.8% (equivalent to 0, 300 and 900 mg/kg bw/day) to 8 animals/sex/dose and via drinking water at 0, 0.07 and 0.2% (equivalent to 0, 85 and 145 mg/kg bw/day to 9 animals/sex/dose. Rats at the 0.6 and 1.8% dose group exhibited severe weight loss so LAS administration via diet was stopped after 2 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. Terminal blood collection was done for estimation of haematological and clinical chemistry parameters. Gross findings were observed after euthanizing animals and organs were removed for organ weight measurements. No histopathology was performed. Body weight gain was reduced at the highest dose group only. Significant decreases in transaminase activity and renal Na,K-ATPase were seen at 145 mg/kg bw/day. No significant haematological or organ weight changes were noted. Based on significant decrease in the liver and renal enzyme levels at 145 mg/kg bw/day, the systemic toxicity NOAEL was established at 85 mg/kg bw/day (Yoneyama, 1976).  

Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
1964
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
A justification of the read-across approach is appended in Section 13.
Reason / purpose:
reference to same study
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
(two dose levels are used in the study against the guideline which recommends at least three dose levels should be used.)
GLP compliance:
no
Remarks:
(pre-GLP)
Limit test:
no
Species:
rat
Strain:
other: FDRL strain (Wistar-derived)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Not specified
- Females nulliparous and non-pregnant: Yes
- Age at study initiation: All animals were at its weaning stage
- Weight at study initiation: Not specified
- Fasting period before study: Not specified
- Housing: Animals were housed individually in raised-bottom, wire-mesh cages
- Diet: Purina laboratory chow, ralston purina co., St. Louis, Missouri; ad libitum
- Water: ad libitum
- Acclimation period: Not specified

No information was provided for food and water quality and environmental conditions.
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
- DIET PREPARATION
No details were provided regarding the preparation of diet. Since rats consume approximately 2.4 times as much diet in proportion to body weight when they are first weaned as when they are adult, lower concentrations were incorporated in their diets initially, and increments in concentration made at biweekly intervals so that the intake of the test materials was relatively constant throughout. By the 11th week on test, when the animals had attained maturity, the levels in the diet were equivalent to 1000 and 5000 ppm of LAS.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
12 weeks
Frequency of treatment:
daily
Dose / conc.:
0.05 other: g/kg bw/day
Dose / conc.:
0.25 other: g/kg bw/day
No. of animals per sex per dose:
15
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Not specified
- Rationale for animal assignment (if not random): Not specified
- Rationale for selecting satellite groups: No satellite group was included in the study.
- Post-exposure recovery period in satellite groups: not specified
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly
- Body weights were recorded for approximately half the rats of each sex.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Food consumption was recorded for approximately half the rats of each sex.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE: Not examined

OPHTHALMOSCOPIC EXAMINATION: Not examined

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During 6th and 12th week of test
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: Half animals from each group (not subjected to body weights and food consumption analysis)
- Parameters: Hemoglobin, hematocrit, total and differential leukocyte counts were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: During 6th and 12th week on test
- Animals fasted: Not specified
- How many animals: Half animals from each group (not subjected to body weights and food consumption analysis)
- Parameters: Blood glucose and urea nitrogen were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: During 6th and 12th week on test
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Half animals from each group (not subjected to body weights and food consumption analysis)
- Parameters: Albumin, pH, glucose, and microscopic examination of the sediment were examined.

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
Following the 12-week clinical examinations, all surviving rats were euthanized by over dosage with sodium pentobarbital, and examined at necropsy for evidence of gross pathologic changes. The liver, kidneys, spleen, heart, adrenals, pituitary, and cecum of each rat were weighed. Representative specimens of all organs and tissues from each animal were preserved in formalin.

Microscopic examinations were made of hematoxylin-eosin stained sections of the liver, spleen, stomach, small intestine, large intestine, pancreas, kidney, bladder, adrenal, gonads, thyroid, pituitary, thymus, salivary gland, lymph node, heart, lung, femoral marrow, aorta, muscle, spinal cord, and brain of 5 rats of each sex in the control and 250 mg/kg bw/day groups. Microscopic examinations were also made of the liver, spleen, stomach, kidneys, and cecum of all animals in the 50 mg/kg bw/day groups.
Clinical signs:
no effects observed
Mortality:
not specified
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
Occasional signs of wetting over the posterior ventral region of were noted during the first 14 days for females fed with 0.25 g/kg bw/day.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The t values for relative liver weight in the females in the 0.25 g/kg bw/day LAS group indicated a statistical significant increase in weight of this organ, compared to the controls. The weights of other organs did not show significant differences from the controls.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The frequent finding at gross necropsy was hemorrhagic areas in the lungs, characteristic of the common type of pneumonitis seen in rats. The scattered incidence of fluid-containing kidneys of rats in some groups was also noticed. However, both of these findings were not associated with the test materials or dosage levels and was unrelated to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
50 other: mg/kg bw/day
Based on:
act. ingr.
Sex:
female
Basis for effect level:
organ weights and organ / body weight ratios
Remarks on result:
other: There was significant increase in relative liver weights in females fed with 250 mg/kg bw/day
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
250 other: mg/kg bw/day
System:
gastrointestinal tract
Organ:
liver
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
yes

Table 1: Weight gain, efficiency of food utilization and sample intake of rats fed alkyl benzene for 12 weeksa. (Oser and Morgareidge, 1965)

Dosageb sex Mean weight gain (g) Efficiency of food utilization (g/100 g)c sample intake (g/kg/day)
Controls M 313 18.8 None
  F 169 13
0.05 g/kg/day M 315 18.6 0.05
  F 164 12.6
0.25 g/kg/day M 314 18.8 0.256
  F 163 12.6

a: The values given are the means of 7 or 8 animals of each sex.

b: These are nominal dosages. The actual dosages, calculated from food consumption, are given

as “sample intake” in the last column.

c: As grams of weight gained per 100 g of food eaten.

Table 2: Organ weights of rats fed alkyl benzene sulfonates in the diet for 12 weeks (Oser and Morgareidge, 1965)

Organ Sex Organ weighta(%)
Controls 0.05 g LAS/kg/day 0.25 g LAS/kg/day
Liver M 3.58 3.81 3.79
F 3.86 3.77 4.34b
Kidneys M 0.7 0.75 0.73
F 0.79 0.8 0.78
Spleen M 0.21 0.25 0.22
F 0.31 0.28 0.3
Heart M 0.36 0.33 0.34
F 0.39 0.4 0.37
Adrenals M 0.013 0.012 0.012
F 0.027 0.029 0.027
Pituitary M 0.004 0.004 0.004
F 0.007 0.006 0.007
Cecum M 0.47 0.49 0.5
F 0.58 0.57 0.58

a: as percentage of bodyweight.

b: Different from corresponding controls of same sex, P < 0.01.

Table3: Gross pathologic findings in rat fed alkyl benzene sulfonates in the diet for 12 weeksa (Oser and Morgareidge, 1965)

Organ and findings Control 0.05 g LAS/kg/day 0.25 g LAS/kg/day
M F M F M F
Cecum: Slight hyperemia - - - 5112 - -
Kidneys: Soft and hollow - - 5098 5106; 5111 5124 5146; 5148
Hollow and filled with fluid - 5022 - 5113 - 5147
Congenital absence of right kidney left; hypertrophied 5009 - - - - -
Adrenals: Hyperemic and/or slightly hemorrhagic 5003;5012 -   5114; 5119 5124; 5125 5145
Thymus: Petecbial hemorrhage - - - 5113 5130 -
Lungs: Hemorrhagic areas 5002 5029 5092 5114; 5115; 5119 5124; 5130 5144; 5150
Purulent leisions - - 5102 - - -
Edema - - - - - 5136
Areas of emphysema - 5030 5091 5110 - -

a: The numbers given are the identifying numbers of the individual animals that exhibited the lesion.

Conclusions:
Administration of sodium linear alkylbenzene sulphonate with nominal chain length of C12 (range C9-15) to albino rats at dose levels of 0.05 and 0.25 g/kg bw/day resulted in a NOAEL of 0.05 g/kg bw/day for female rats, based on increased relative liver weights and 0.25 g/kg bw/day for male rats, based on no abnormal variations in growth, food intake, hematology, clinical biochemistry, urinalysis, gross pathology, organ weights and histopathology in any of the groups observed
Executive summary:

A sub-chronic repeated dose dietary toxicity study was conducted with C10-14 LAS, sodium salt in Wistar FDRL rats. The test substance was administered to groups of 5 male and 5 female rats at dietary dose levels of 0, 50 and 250 mg/kg bw/day. Appearance, behaviour and overt signs of toxicity were observed daily. Food consumption and body weights of the rats of were recorded once weekly. During 6th and 12th week on test haematological, clinical chemistry and urine parameters were analysed. Following the 12-week clinical examinations, all surviving rats were sacrificed and gross pathological examination was performed. The major organs were weighed and histopathology was performed. No adverse effects on body weight gain, food consumption and behaviour were observed. The clinical data showed no abnormal variations in any of the dose groups. The relative organ weights and the histopathological evaluation did not show significant differences among the dose groups except for a significant liver weight increase in females of the highest dose group. Under the study conditions, the systemic toxicity NOAEL was established at 50 mg/kg bw/day based on increased relative liver weights in the females at 250 mg/kg bw/day (Oser, 1965).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
85 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Organ:
kidney
liver

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Reason / purpose:
reference to same study
Principles of method if other than guideline:
The purpose of this study is to determine the chronic toxicity (transdermal) of C10 - C13 linear alkylbenzene sulfonic acid magnesium salt (LAS-Mg) in CRJ-SD rats. Male and female rats were transdermally administered with 0 (water), 3% PEG, 0.5, 1.0 and 5.0% of test substance dissolved in 3% polyethyleneglycol (PEG, MW: 200) for 26 weeks. General symptoms and body weight of animals were recorded on a daily-basis and food intake were recorded weekly. Urinalysis and hematological examinations were performed in week 5 and 13 of the study. At the end of study, urinalysis, haematological and serum biochemistry were also done. All the surviving animals were humanely euthanized after 26 weeks and organs (liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary, brain and thyroid) were collected for recording absolute and relative organ weights. All the collected organs were examined under the microscope for histopthalogical changes.
GLP compliance:
no
Remarks:
(pre-dates GLP)
Limit test:
no
Species:
rat
Strain:
other: CRJ-SD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Male and female rats were obtained from Charles River Laboratories Japan, Inc.
- Age at study initiation: Approximately 4 weeks
- Weight at study initiation: Not reported
- Fasting period before study: Not reported
- Housing: Animals were housed in cages with 2 animals/cage.
- Diet: Solid feed (CE-2, CLEA Japan, Inc.); ad libitum
- Water: Tap water; ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 1°C
- Humidity: 55 ± 5%
- Air changes: Not reported
- Photoperiod: Not reported

IN LIFE DATES: Not reported
Type of coverage:
not specified
Vehicle:
other: 3% polyethyleneglycol (PEG, MW: 200)
Details on exposure:
TEST SITE
- Area of exposure: Shaved backs of animals
- % coverage: 3 x 3 cm2 area in the middle of the backside
- Type of wrap if used: Not reported
- Time intervals for shavings or clippings: Weekly

REMOVAL OF TEST SUBSTANCE
- Washing: Not reported
- Time after start of exposure: Not reported

TEST MATERIAL
- Amount(s) applied: 0.1 mL
- Concentration: 0.5, 1.0 and 5.0% of C10-C13 LAS-Mg in 3% polyethyleneglycol

VEHICLE
- 3% polyethyleneglycol (PEG, MW: 200)

USE OF RESTRAINERS FOR PREVENTING INGESTION: No
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
26 weeks
Frequency of treatment:
Once daily for 26 weeks (excluding Sundays)
Dose / conc.:
0.5 other: %
Dose / conc.:
1 other: %
Dose / conc.:
5 other: %
No. of animals per sex per dose:
20 animals/sex/dose group
Control animals:
other: 2 Control animals groups were applied with either PEG or distilled water.
Details on study design:
- Dose selection rationale: Not reported
- Rationale for animal assignment: Not reported
- Rationale for selecting satellite groups: No satellite groups were used in the study
Positive control:
Not included
Observations and examinations performed and frequency:
CLINICAL OBSERVATIONS: Yes
- Time schedule: All animals were observed daily for general symptoms.

DERMAL IRRITAION: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: All animals were weighed daily.

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule: Amount of food intake was measured once per week.

FOOD EFFICIENCY: Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During weeks 5, 11 and 26
- Anaesthetic used for blood collection: Yes. The animals were anesthetized with sodium pentobarbital
- Animals fasted: Not reported
- How many animals: During weeks 5 and 11, blood was collected from 10 animals/sex from PEG group, control group and 5.0% group and in week 26, blood was collected from all animals.
- Parameters checked: Red blood cell count, white blood cell count, hemoglobin level and hematocrit level were measured, and white blood cell fractions were calculated by Giemsa-staining the blood smear samples.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At end of study i.e. 26 weeks
- Animals fasted: Not reported
- How many animals: All animals
- Parameters checked: s-GOT, s-GPT, alkaline phosphatase, total protein, albumin, glucose, cholesterol, bilirubin, creatinine, urea nitrogen, inorganic phosphates and Cl-, Na+, K+, Ca2+ and Mg2+

URINALYSIS: Yes
- Time schedule for collection of urine: During weeks 5, 11 and 26
- Metabolism cages used for collection of urine: No
- Animals fasted: Not reported
- How many animals: During weeks 5 and 11, urine was collected from 10 animals/sex from PEG group, control group and 5.0% group and in week 26, urine was collected from all animals.
- Method: Lab sticks, Urobilistix and iktstix method (Semi-quantitative investigation)
- Parameters: pH, protein, glucose, ketone bodies, occult blood, urobilinogen and bilirubin

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals were euthanized after blood sampling and organs were immediately collected.

TISSUE COLLECTED FOR ORGAN WEIGHTS AND HISTOPATHOLOGY: Liver, kidneys, spleen, adrenal glands, lungs, heart, thymus, reproductive organ, pituitary, brain and thyroid

HISTOPATHOLOGY: Above collected organs and stomach, duodenum, ileum, cecum, pancreas, bone marrow and skin were fixed in 10% formalin, embedded in paraffin and underwent H&E staining before being examined under the microscope
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
In half of the animals of highest tested dose, mild redness of test site was observed one week after starting to apply it, but this disappeared after one week with desquamation. This did not occur again thereafter. Among female animals, all animals in the 5.0% group and one animal from the 1.0% group developed mild redness after one month since the start of study, but these went away after one week. After two, three months, half the animals in the 5.0% group exhibited mild redness, and it was seen sporadically thereafter.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One male animal from the control group (Week 16) and one male animal from the 5.0% group (Week 23) died during the study, but the cause remains unknown after finding no abnormalities through the necropsies.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Male animals in the 0.5% group exhibited mild weight suppression from Week 11, but this had recovered by Week 16. At the end of the experiment, slight weight suppression was observed in male animals from the 0.5% group.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
5 weeks interim analysis: There were significant increase in segmented neutrophils and significant decrease in white blood cell count for male animals in PEG group and increase in hematocrit levels in male animals in 5.0% dose group
13 weeks interim analysis: level: Significant increase in red blood cells and hemoglobin levels were observed in males of 5.0% dose group and increased white blood cell count was also observed for male and female animals in the PEG and 5.0% groups. In white blood cell fractions, decrease in eosinophils was observed for female animals in the 5.0% group.
6 month analysis: Significant decrease in white blood cell count were observed for female animals in the 5.0% group, as well as for both male and female animals in the 1.0% group. Significant increase in lymphocytes for male animals in the PEG group and significant decrease in monocytes for male animals in the 5.0 and 0.5% groups were also observed.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
PEG group: Significant decrease was observed in s-GOT levels for female animals and in sodium and s-GPT levels for all animals.
5.0% dose group: There was significant reduction in glucose in females, in calcium levels in males and sodium and s-GPT in all animals. Elevated levels of A/G ratio and cholesterol were also observed in females.
1.0% dose group: There is significant decrease in glucose in females and sodium levels in all animals and increase in urea nitrogen in females of this group.
0.5% dose group: There is significant decrease in sodium levels in all animals of this group and increase in urea nitrogen in females of this group.
Urinalysis findings:
no effects observed
Description (incidence and severity):
Similar levels of all tested parameters (protein, occult blood, Urobilinogen and pH) were observed in treated animals as compare to control animals in 13 as well as 26 weeks interim analysis.
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
There were decrease in actual and relative liver weights of 0.5% (low) dose group. No dose dependency was observed for this effect.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
- Liver stasis was observed for one male animal in the 1.0% group, while blood spots and fading color were observed for another male animal in the same group. Furthermore, white serpentine bulges were observed for three male animals each in the 5.0 and 0.5% groups, as well as in four male animals in 1.0% group and one male animal in the 0.5% group.
- Kidney edema was observed for one female each in the control and 0.5% groups.
- Yellow discoloration was observed in pancreas for one male animal in the control group.
- Lungs stasis was observed for one male animal in the 1.0% group.
- Heart hypertrophy was observed for one male animal in the 1.0% group.
- Blood spots in thymus were observed for two male animals each in the control, PEG and 1.0% groups, as well as in four male animals in the 5.0 and 0.5% groups.
- Testicular atrophy was observed for one male in the 1.0% group, while ovarian edema was observed for one female animal each in the control and 1.0% groups.
- Pituitary edema was observed for one male animal in the 1.0% group.
- Stomach hematoma was observed in the cardia for one male animal in the 1.0% group, while an ulcer of 1 mm in diameter was observed for another male animal in the same group. Edema was observed in the stomach for one male animal in the control group.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Control group:
- Sinusoid bleeding was observed for one male and one female animal each, while focal necrosis was observed for one female animal.
- Renal tubular hyaline casts were observed for seven male animals and one female animal, while interstitial small round cell infiltration and renal tubular epithelial cell swelling were observed for two male and one female animals
- Fibrosis of islets of Langerhans observed for one male animal
- Interstitial myocarditis was observed for one male and one female animal each.
PEG group:
- Sinusoid bleeding was observed for three male animals, focal necrosis was observed in one male animal and Kupffer cell migration was observed for one female animal
- Renal tubular hyaline casts were observed for five male animals and one female animal, while interstitial small round cell infiltration and renal tubular epithelial cell swelling were observed for one male and one female animal each
- Adrenal gland: A hemorrhagic foci was observed in the stratum corneum for one male animal.
- Fibrosis of islets of Langerhans observed for one male animal.
5.0% dose group:
- Sinusoid bleeding was observed for five male animals, while focal necrosis was observed for two male animals. Kupffer cell migration was observed for one female animal.
- Renal tubular hyaline casts were observed for three male and female animals each, while interstitial small round cell infiltration was observed for two male animals. Renal tubular calcification was observed for another male animal.
- Fibrosis of islets of Langerhans observed for one male animal.
- Interstitial myocarditis was observed for two male animals and one female animal.
1.0% dose group:
- Sinusoid bleeding was observed for five male animals, while focal necrosis was observed for another male animal.
- Renal tubular hyaline casts were observed for five male and one female animals, interstitial small round cell infiltration was observed for one male and one female animal each, and renal tubular calcification was observed for two female animals
- Fibrosis of islets of Langerhans observed for four male animals
- Interstitial myocarditis was observed for one male animal.
- Decline in sperm production was observed for one male animal.
0.5% dose group:
- Sinusoid bleeding was observed for four male animals and one female animal, while large lipid droplets were observed for one male animal. Kupffer cell migration was observed for one female animal.
- Renal tubular hyaline casts were observed for five male and two female animals, interstitial small round cell infiltration was observed for one male animal and another male animal exhibited renal tubular epithelial cell swelling. Furthermore, renal tubular calcification was observed for one female animal.
- Fibrosis of islets of Langerhans observed for three male animals.
- Interstitial myocarditis was observed for three male animals.
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
- In half of the animals of highest tested dose, mild redness of test site was observed one week after starting to apply it, but this disappeared after one week with desquamation. This did not occur again thereafter.
- Among female animals, all animals in the 5.0% group and one animal from the 1.0% group developed mild redness after one month since the start of study, but these went away after one week.
- After two, three months, half the animals in the 5.0% group exhibited mild redness, and it was seen sporadically thereafter.
- One male animal from the control group (Week 16) and one male animal from the 5.0% group (Week 23) died during the study, but the cause remains unknown after finding no abnormalities through the necropsies.

BODY WEIGHT AND WEIGHT GAIN
- Male animals in the 0.5% group exhibited mild weight suppression from Week 11, but this had recovered by Week 16.
- At the end of the experiment, slight weight suppression was observed in male animals from the 0.5% group.

FOOD CONSUMPTION AND COMPOUND INTAKE
- No significant difference was noted in males or females between the control and test substance groups.

FOOD EFFICIENCY
- No effects were observed in test substance treated animals as compare to control animals

HAEMATOLOGY
- 5 weeks interim analysis: There were significant increase in segmented neutrophils and significant decrease in white blood cell count for male animals in PEG group and increase in hematocrit levels in male animals in 5.0% dose group
- 13 weeks interim analysis: level: Significant increase in red blood cells and hemoglobin levels were observed in males of 5.0% dose group and increased white blood cell count was also observed for male and female animals in the PEG and 5.0% groups. In white blood cell fractions, decrease in eosinophils was observed for female animals in the 5.0% group.
- 6 month analysis: Significant decrease in white blood cell count were observed for female animals in the 5.0% group, as well as for both male and female animals in the 1.0% group. Significant increase in lymphocytes for male animals in the PEG group and significant decrease in monocytes for male animals in the 5.0 and 0.5% groups were also observed.

CLINICAL CHEMISTRY
- PEG group: Significant decrease was observed in s-GOT levels for female animals and in sodium and s-GPT levels for all animals.
- 5.0% dose group: There was significant reduction in glucose in females, in calcium levels in males and sodium and s-GPT in all animals. Elevated levels of A/G ratio and cholesterol were also observed in females.
- 1.0% dose group: There is significant decrease in glucose in females and sodium levels in all animals and increase in urea nitrogen in females of this group.
- 0.5% dose group: There is significant decrease in sodium levels in all animals of this group and increase in urea nitrogen in females of this group.

URINALYSIS
Similar levels of all tested parameters (protein, occult blood, Urobilinogen and pH) were observed in treated animals as compare to control animals in 13 as well as 26 weeks interim analysis.
ORGAN WEIGHTS
- There were decrease in actual and relative liver weights of 0.5% (low) dose group. No dose dependency was observed for this effect.

GROSS PATHOLOGY
- Liver stasis was observed for one male animal in the 1.0% group, while blood spots and fading color were observed for another male animal in the same group. Furthermore, white serpentine bulges were observed for three male animals each in the 5.0 and 0.5% groups, as well as in four male animals in 1.0% group and one male animal in the 0.5% group.
- Kidney edema was observed for one female each in the control and 0.5% groups.
- Yellow discoloration was observed in pancreas for one male animal in the control group.
- Lungs stasis was observed for one male animal in the 1.0% group.
- Heart hypertrophy was observed for one male animal in the 1.0% group.
- Blood spots in thymus were observed for two male animals each in the control, PEG and 1.0% groups, as well as in four male animals in the 5.0 and 0.5% groups.
- Testicular atrophy was observed for one male in the 1.0% group, while ovarian edema was observed for one female animal each in the control and 1.0% groups.
- Pituitary edema was observed for one male animal in the 1.0% group.
- Stomach hematoma was observed in the cardia for one male animal in the 1.0% group, while an ulcer of 1 mm in diameter was observed for another male animal in the same group. Edema was observed in the stomach for one male animal in the control group.

HISTOPATHOLOGY
Control group:
- Sinusoid bleeding was observed for one male and one female animal each, while focal necrosis was observed for one female animal.
- Renal tubular hyaline casts were observed for seven male animals and one female animal, while interstitial small round cell infiltration and renal tubular epithelial cell swelling were observed for two male and one female animals
- Fibrosis of islets of Langerhans observed for one male animal
- Interstitial myocarditis was observed for one male and one female animal each.
PEG group:
- Sinusoid bleeding was observed for three male animals, focal necrosis was observed in one male animal and Kupffer cell migration was observed for one female animal
- Renal tubular hyaline casts were observed for five male animals and one female animal, while interstitial small round cell infiltration and renal tubular epithelial cell swelling were observed for one male and one female animal each
- Adrenal gland: A hemorrhagic foci was observed in the stratum corneum for one male animal.
- Fibrosis of islets of Langerhans observed for one male animal.
5.0% dose group:
- Sinusoid bleeding was observed for five male animals, while focal necrosis was observed for two male animals. Kupffer cell migration was observed for one female animal.
- Renal tubular hyaline casts were observed for three male and female animals each, while interstitial small round cell infiltration was observed for two male animals. Renal tubular calcification was observed for another male animal.
- Fibrosis of islets of Langerhans observed for one male animal.
- Interstitial myocarditis was observed for two male animals and one female animal.
1.0% dose group:
- Sinusoid bleeding was observed for five male animals, while focal necrosis was observed for another male animal.
- Renal tubular hyaline casts were observed for five male and one female animals, interstitial small round cell infiltration was observed for one male and one female animal each, and renal tubular calcification was observed for two female animals
- Fibrosis of islets of Langerhans observed for four male animals
- Interstitial myocarditis was observed for one male animal.
- Decline in sperm production was observed for one male animal.
0.5% dose group:
- Sinusoid bleeding was observed for four male animals and one female animal, while large lipid droplets were observed for one male animal. Kupffer cell migration was observed for one female animal.
- Renal tubular hyaline casts were observed for five male and two female animals, interstitial small round cell infiltration was observed for one male animal and another male animal exhibited renal tubular epithelial cell swelling. Furthermore, renal tubular calcification was observed for one female animal.
- Fibrosis of islets of Langerhans observed for three male animals.
- Interstitial myocarditis was observed for three male animals.
Key result
Dose descriptor:
NOAEL
Effect level:
5 other: %
Based on:
act. ingr.
Remarks:
corresponding to 2500 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: No significant adverse effect was seen at any dose level.
Remarks on result:
other: Based on no growth, functional and morphological abnormalities, other than the localized effects at any tested dose levels.
Key result
Critical effects observed:
no
Conclusions:
Transdermal administration of C10-13 magnesium linear alkylbenzene sulphonate (LAS-Mg; purity: 96.9%) to CRJ-SD rats at dose levels of 0 (water), 3% PEG, 0.5, 1.0 and 5.0% of test substance dissolved in 3% polyethyleneglycol (PEG, MW: 200) for 26 weeks revealed no observed adverse effect level (NOAEL) of 5.0%, based no toxicologically relevant changes at any dose level.
Executive summary:

A sub-chronic 26-week repeated dose toxicity study was conducted in male and female CRJ-SD rats to evaluate the effects of C10-13 LAS, magnesium salt. The test substance was applied daily on the shaved backs of animals at dose levels of 0, 0.5, 1.0 and 5.0% dissolved in 3% polyethylene glycol (PEG) for 26 weeks with 20 animals/sex/dose group. General symptoms and body weight of animals were recorded on a daily-basis and food intake were recorded weekly. Urinalysis and haematological examinations were performed on Weeks 5 and 13 and also at termination, along with clinical biochemical examination. All the surviving animals were humanely euthanized after 26 weeks and organs were collected for recording absolute and relative weights and histopathological observation. There was slight reduction of body weight in males at 0.5% at the end of the study. No toxicologically significant changes were observed in food consumption, urinalysis, haematological and serum biochemical findings. No changes were seen in the organ weights and gross pathology of the collected organs. Histopathological findings were non-specific and had no dose-dependency. Under the study conditions, the systemic toxicity NOAEL was established at 5.0% (equivalent to 2500 mg/kg bw/day), based no toxicologically relevant changes at any dose level (Ito, 1978d). 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
2 500 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Oral 

  

Study 1 (C10-14LAS): 

A sub-chronic (9 months) repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in male and female Wistar JCL rats. The test substance was administered at the following dose levels: through diet at 0, 0.6 and 1.8% (equivalent to 0, 300 and 900 mg/kg bw/day) to 8 animals/sex/dose and via drinking water at 0, 0.07 and 0.2% (equivalent to 0, 85 and 145 mg/kg bw/day to 9 animals/sex/dose. Rats at the 0.6 and 1.8% dose group exhibited severe weight loss so LAS administration via diet was stopped after 2 weeks. Clinical observations, water consumption, food consumption and body weights were recorded weekly. Terminal blood collection was done for estimation of haematological and clinical chemistry parameters. Gross findings were observed after euthanizing animals and organs were removed for organ weight measurements. No histopathology was performed. Body weight gain was reduced at the highest dose group only. Significant decreases in transaminase activity and renal Na,K-ATPase were seen at 145 mg/kg bw/day. No significant haematological or organ weight changes were noted. Based on significant decrease in the liver and renal enzyme levels at 145 mg/kg bw/day, the systemic toxicity NOAEL was established at 85 mg/kg bw/day (Yoneyama, 1976).

  

Study 2 (MEA): 

  

Effects of repeated exposure to MEA was evaluated in the context of a two-generation reproductive toxicity study according to OECD Guideline 416 in rats. The test substance was administered to 25 rats/sex/dose via diet at doses of 0, 100, 300 and 1,000 mg/kg bw/day (nominal) through two successive generations. Clinical examination revealed no test substance-related adverse effects in F0 and F1 parental animals of low and mid doses. The test substance adversely affected food consumption at the high dose F0 females during lactation. Also, the body weight gain and, for F0 generation, body weights of the high-dose dams were statistically significantly less compared to the control group during gestation, which was likely be secondary to an increased post-implantation loss in these animals. Regarding the gross pathological examinations, the decrease of absolute weights of cauda epididymis, epididymis, and prostate in F0 and F1 male top-dose animals were considered as treatment-related effects. There were no other treatment-related systemic effects in any of the generations. Under the study conditions, the systemic toxicity NOAEL of MEA in rats was determined to be 300 mg/kg bw/day (ACC and CEFIC, 2009).

Study 3 (C10-14LAS): 

  

A sub-chronic repeated dose toxicity study was conducted with C10-14 LAS, sodium salt in Wistar SLC rats. The test substance was administered to groups of 10 male and female rats/dose in the diet for 26 weeks at doses of 0, 40, 115, 340 and 1030 mg/kg/day. Clinical observations, water consumption, food consumption and body weights were recorded weekly. At the end of study, haematological, clinical chemistry and urine parameters were analysed. Gross pathological examinations were observed after necropsy and the major organs were subjected to histopathological examinations. Significant diarrhoea, suppressed growth, increased caecal weight and degeneration of renal tubes were observed in the highest dose group. Similar but less severe signs were seen in other doses with the exception of the lowest dose of 40 mg/kg/day, which showed no adverse effects related to exposure to the LAS. Under the study conditions, systemic toxicity NOAEL was determined to be 40 mg/kg/day, based on tissue damage in caecum, kidney and liver, haematological as well as clinical chemistry parameters changes at higher dose levels (Yoneyama, 1972)

  

Study 4 (C10-13LAS): 

  

A short-term repeated dose toxicity study was conducted with C10-13 LAS, sodium salt in CRJ-SD rats. The test substance was administered via oral gavage to groups of male and female rats for 28 days at dose levels of 0, 125, 250 and 500 mg/kg/day. Clinical signs of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters, following which the animals were humanely euthanized, gross necropsy was performed, the organs weights were determined and histopathological examination of the major organs were performed. Body weight gain was suppressed, some serum biochemical measures were different from the controls, and some organ weights were either decreased (spleen, heart, thymus) or increased (liver) at the high dose. No mortalities or histopathological abnormalities were observed. No adverse effects were up to 125 mg/kg bw/day mg/kg/day. Under the study conditions, the systemic toxicity NOAEL was determined to be 125 mg/kg/day based on reduced body weight, changes in organ weight (without histopathological correlation) and clinical chemistry at the mid and high doses (Ito, 1978a). 

  

Study 5 (C10-14LAS): 

  

A sub-chronic repeated dose dietary toxicity study was conducted with C10-14 LAS, sodium salt in Wistar FDRL rats. The test substance was administered to groups of 5 male and 5 female rats at dietary dose levels of 0, 50 and 250 mg/kg bw/day. Appearance, behaviour and overt signs of toxicity were observed daily. Food consumption and body weights of the rats of were recorded once weekly. During 6th and 12th week on test haematological, clinical chemistry and urine parameters were analysed. Following the 12-week clinical examinations, all surviving rats were sacrificed and gross pathological examination was performed. The major organs were weighed and histopathology was performed. No adverse effects on body weight gain, food consumption and behaviour were observed. The clinical data showed no abnormal variations in any of the dose groups. The relative organ weights and the histopathological evaluation did not show significant differences among the dose groups except for a significant liver weight increase in females of the highest dose group. Under the study conditions, the systemic toxicity NOAEL was established at 50 mg/kg bw/day based on increased relative liver weights in the females at 250 mg/kg bw/day (Oser, 1965).

  

Study 6 (C10-13LAS): 

  

A sub-chronic repeated dose toxicity study was conducted with C10-13LAS, magnesium salt in CRJ-SD rats. The test substance was administered daily at dose levels of 0, 75, 150 and 300 mg/kg bw/day via oral gavage to groups of 20 animals/sex for 26 weeks. General symptoms and body weight of animals were recorded on a daily basis and food intake was recorded weekly. Urinalysis, haematological and clinical biochemistry examinations were performed during the in-life phase as well as at termination. Following necropsy of the surviving animals, gross pathological and histopathological examinations were performed. There was slight body weight suppression in the males at 300 mg/kg bw/day. Haematological findings revealed significant reduction in haematocrit levels for females at 300 mg/kg bw/day. There was significant reduction in bilirubin, protein, albumin and calcium levels in male animals and significant reduction in SGPT, glucose, calcium and magnesium levels in female animals at 300 mg/kg bw/day. Reduction in relative heart weight (females), kidney weight (males) and adrenal gland weight (females) were also observed in the highest dose group. In the kidneys, renal tubular epithelial cell swelling, hyaline casts and interstitial small round cell infiltration were observed at 300 mg/kg bw/day. There were no toxicologically significant effects at the lower doses. Under the study conditions, the systemic toxicity NOAEL was established at 150 mg/kg bw/day, based on reduction in body weight, fluctuations in haematological and serum biochemistry, altered organ weights and histopathological alterations in kidney at 300 mg/kg bw/day (Ito, 1978b).  

  

Study 7 (C10-13LAS): 

  

A sub-acute repeated dose toxicity study was conducted with C10-13LAS, magnesium salt in CRJ-SD rats. The test substance was administered to groups of male and female rats via oral gavage at doses of 0, 155, 310 and 620 mg/kg bw/day for 29/30 days. Clinical signs of toxicity and the body weights were recorded daily while the food consumption was measured twice a week. Urinalysis was also performed during the study. At the end of study, blood was collected for estimation of haematological and clinical chemistry parameters, following which the animals were humanely euthanized, gross necropsy was performed and the organs weights were determined for the major organs. Based on the results of organ weight measurements, histopathology was performed only for the livers of male animals from the test and control groups. 1 male and 2 female animals died at the highest dose. The relative liver weight of male and female animals was significantly increased at 620 mg/kg bw/day. Apart from this, increase in relative testes weight and brain weight were also observed at this dose. Decrease in the relative heart weight (at 310 and 620 mg/kg bw/day) and thymus (at 620 mg/kg bw/day) was observed in the female animals. Although, some significant effects were observed at low and mid doses yet they were not dose-dependent. Histopathology revealed focal necrosis and increased glycogen levels in the liver of few animals belonging to the 155 and 310 mg/kg bw dose groups but these findings were comparable to the control group and were not dose-dependent. Under the study conditions, the systemic toxicity NOAEL was established at 310 mg/kg bw/day, based on mortality, reduced body weight and changes in organ weight (without biochemical or histopathological correlation) at the higher dose level (Ito, 1978c).

 

Study 8 (MEA): 

  

A sub-acute repeated dose toxicity study was conducted with monoethanolamine (MEA) in Sprague-Dawley rats. The test substance was administered to the rats through diet at dose levels of 0%, 0.01%, 0.1% and 1% (equivalent to 0, 7.7, 77 and 770 mg/kg bw/day) for a period of 32 days during which individual body weight and group feed consumption were recorded. On Day 28, haematological examination was performed on the control and high dose group animals. Following sacrifice of the surviving animals, gross pathology of all organs and weight of liver, kidneys, spleen, heart, lung, brain, and testes were recorded. No significant difference in body weight and feed consumption were observed. There was also no difference found in the mean haematogram of the high dose rats. No treatment-related effects were observed in the organ weights, gross pathology and histopathology of the organs. Under the study conditions, the systemic toxicity NOAEL was established at dietary level of 1% (770 mg/kg bw/day) based on absence of effects in body weight gain, feed efficiency, hematologic, organ weights, pathological and microscopic findings (Terhaar, 1967).  

  

Study 9 (MEA): 

  

A chronic repeated dose toxicity was conducted with monoethanolamine (MEA) in beagle dogs. The test substance as a hair dye composite (containing 22.42% MEA) was administered in the diet to groups of 3 male and 3 female dogs for 2 years at concentrations of 0, 19.5, and 97.5 mg/kg bw/day. Apart from regular observation of clinical signs of toxicity, body weight and feed consumption, physical examinations including funduscopic, EKG, blood pressure, pulse rate, and body temperature determinations were conducted initially and at 3, 6, 12, 18, and 24 months. Haematological, clinical biochemistry and urinalysis parameters were determined for the high dose and control groups and on three male and three female dogs in the low dose group at the same times. Necropsy was performed on one male and one female from each group at 6,12, and 18 months, and on all survivors at 24 months and organ weight determination, gross pathology and histopathology were performed. No treatment-related adverse effects were observed in any of the parameter studied between controls and animals at 19.5 or 97.5 mg/kg bw/day. Under the study conditions, the systemic toxicity NOAEL was established at ≥97.5 mg/kg bw/day (equivalent to 20.5 mg a.i. monoethanolamine/kg bw/day), based on absence of statistically significant effects in the treated animals (Wernick, 1975).  

  

Study 10 (MEA): 

  

A range finding repeated dose toxicity test was performed with monoethanolamine (MEA) tudy in rats. The test substance was administered via diet to groups of ten rats at doses in the range of 160-2670 mg/kg bw/ day for 90 days. No reduced growth and food consumtpnio were observed in animals. Altered kidney and liver weight were observed at dose of 640 mg/kg bw/day. Microscopic lesions and mortality were also observed at 1280 mg/kg bw/day. Under the study conditions, the systemic toxicity NOAEL was established at 320 mg/kg bw/day (Smyth, 1951). 

 


Dermal 

  

Study 1 (C10 -13LAS): 

  

A sub-chronic 26 -weeks repeated dose toxicity study was conducted in male and female CRJ-SD rats to evaluate the effects of C10-13LAS, magnesium salt. The test substance was applied daily on the shaved backs of animals at dose levels of 0, 0.5, 1.0 and 5.0% dissolved in 3% polyethylene glycol (PEG) for 26 weeks with 20 animals/sex/dose group. General symptoms and body weight of animals were recorded on a daily-basis and food intake were recorded weekly. Urinalysis and haematological examinations were performed on Weeks 5 and 13 and also at termination, along with clinical biochemical examination. All the surviving animals were humanely euthanized after 26 weeks and organs were collected for recording absolute and relative weights and histopathological observation. There was slight reduction of body weight in males at 0.5% at the end of the study. No toxicologically significant changes were observed in food consumption, urinalysis, haematological and serum biochemical findings. No changes were seen in the organ weights and gross pathology of the collected organs. Histopathological findings were non-specific and had no dose-dependency. Under the study conditions, the systemic toxicity NOAEL was established at 5.0% (equivalent to 2500 mg/kg bw/day), based no toxicologically relevant changes at any dose level (Ito, 1978d).

 

  

  

Other routes 

 

Study 1 (C10-13 LAS):

A 28-day repeated dose toxicity was conducted with C10-13LAS, sodium salt in Rhesus monkeys via oral and subcutaneous routes. The test substance was administered to the monkeys at doses of 0, 30, 150 and 300 mg/kg bw/day and at doses of 0, 0.1, 0.5 and 1.0 mg/kg bw/day for 28 days by simultaneous oral gavage and subcutaneous injection (into the dorsal aspect of the upper thorax) respectively. Clinical observations for any signs of toxicity, water consumption and food consumption were observed daily, and body weights was recorded once weekly. Ophthalmological, haematological, clinical chemistry and urine parameters were analysed before the start of the study and during the final week of dosing. Organ weights, gross pathological and histopathological examinations were performed on the major organs. No significant adverse effect was observed at food and water consumption of animals. All animals at the highest dose vomited on numerous occasions, usually within 3 h of being dosed. Vomiting was often associated with salivation. These effects are probably related to the inherent irritative effects of LAS rather than to its systemic toxicity. There was no further evidence of treatment-related effects among the ophthalmological, laboratory and other pathological investigations performed during this study. There was also a dose-related increase in the occurrence of chronic inflammatory cell infiltration (mainly fibroblasts) at the subcutaneous injection sites. Fibrosis of the injection sites was found among the entire test group, the incidence and severity being dose-related. Under the study conditions, a systemic toxicity NOAEL of 150 mg/kg bw/day (oral) and 0.5 mg/kg bw/day (sc) (Heywood, 1978). 

Justification for classification or non-classification

Repeated dose studies were conducted with the dissociation products of MEA-LAS (i.e. MEA and LAS) in rats, dogs and monkeys (via oral route), in rats (via dermal route) and in monkeys (via subcutaneous route). No significant effects were observed at the sub-chronic NOAEL of 85 mg/kg bw/day in the 9 months oral gavage study (Yoneyama, 1976) with LAS. Slight to moderate histopathological lesions were observed at the LOAEL of 115 mg/kg bw/day (Yoneyama, 1972) which is above the guidance value recommended in the CLP guidance. Higher systemic NOAEL of 150 mg/kg bw/day were established in the other sub-chronic study (Ito, 1978b). Further, in the ACC/CEFIC, 2009 study with MEA did not show significant effects up to the dose level of 300 mg/kg bw/day. Therefore, based on the results of the repeated dose studies with the dissociation products of MEA-LAS (MEA and LAS), no classification is warranted for this endpoint according to EU CLP (1272/2008/EC) criteria.