Registration Dossier

Administrative data

Endpoint:
chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable study, followed scientific principles/standards, pre-dates-GLP.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Title:
Unnamed
Year:
1978

Materials and methods

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

Test material

Reference
Name:
Unnamed
Test material form:
not specified
Details on test material:
Name of test material: C10-13 magnesium linear alkylbenzene sulphonate (LAS-Mg)
Appearance: White pulverulent body
No further details on test substance are provided in the study report.

Test animals

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

Administration / exposure

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)
Doses / concentrationsopen allclose all
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

Examinations

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

Results and discussion

Results of examinations

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.

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
5 other: %
Based on:
act. ingr.
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.

Applicant's summary and conclusion

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 26-weeks repeated dose toxicity study (transdermal) was conducted in male and female CRJ-SD rats to evaluate the chronic effects of C10-13 magnesium linear alkylbenzene sulphonate (LAS-Mg; purity: 96.9%).

Approximately 4 weeks old male and females animals were housed in cages (2 animals/cage). Solid feed (CE-2, CLEA Japan, Inc.) and tap water were provided ad libitum. The animals were maintained under standard laboratory conditions (temperature: 22 ± 1°C; relative humidity: 55 ± 5%). Prior to the treatment, animals were acclimatized under laboratory conditions for 1 week.

The test substance was administered daily at 5 dose levels of i.e. 0 (water), 3% PEG, 0.5, 1.0 and 5.0% of test substance dissolved in 3% polyethyleneglycol (PEG, MW: 200) 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 hematological examinations were performed in week 5 and 13 of the study.  At the end of study, urinalysis, haematological, 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 and skin) were then fixed in 10% formalin, embedded in paraffin and underwent H&E staining before being examined under the microscope.

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. There were mild redness of test site at Week 1 (in 5.0% dose) and after one month of start of study (in the 5.0% group and one animal from the 1.0% group), but these went away after one week. There was slight weight suppression in male animals from the 0.5% group at the end of the experiment. No toxicologically significant changes were observed in food efficiency of treated animals when compared to controls.

There were no toxicologically relevant changes to the urine, hematological and serum biochemical findings.

No changes were seen in the organ weights of treated animals, and gross necropsy reveal only white serpentine bulges being seen in the livers of male animals in the control and test groups. Histopathological findings were non-specific and had no dose-dependency.

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.