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Toxicological information

Repeated dose toxicity: oral

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Administrative data

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
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1972

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Remarks:
(powder)
Details on test material:
- Name of test material: Linear alkylbenzene sulfonic acid sodium salt (LAS); NeoPlex F-60
- Average molecular weight: 345.8 g
- Composition:: 60 ± 2% (active ingredient) of C10 - C14 LAS, <1.5% unreacted lipids, <4% inorganic salts and 38% moisture
- Lot no.: 127 (Supplier: Kao chemicals)
- Distribution of carbon count of the alkyl groups and component of each sample:
LAS: C10 (10.6%), C11 (34.1%), C12 (27.7%), C13 (19.0%) and C14 (8.7%)

No further details on test substance are provided in the study report.

Test animals

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

Administration / exposure

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)
Doses / concentrationsopen allclose all
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

Examinations

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.

Results and discussion

Results of examinations

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"

Effect levels

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).

Target system / organ toxicity

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

Any other information on results incl. tables

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

Applicant's summary and conclusion

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).