Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: other route
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1977
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1978
Report Date:
1978

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
A 28-day repeated toxicity study was conducted with C10-13 sodium linear alkylbenzene sulphonate (LAS-Na) using rhesus monkeys. Males and female monkeys were administered with 0, 30, 150 and 300 mg/kg bw/day of LAS by oral gavage and simultaneously with 0, 0.1, 0.5 and 1.0 mg/kg bw/day of LAS by subcutaneous injection for 28 days. Control monkeys received distilled water orally and sterile isotonic saline subcutaneously. Clinical observations, water consumption, food consumption and body weights were recorded regularly throughout the study. During the final week of administration, ophthalmological, hematological, clinical chemistry and urine parameters were analyzed for all animals. Gross findings were obtained after euthanizing animals and organs were removed for organ weight measurements.
GLP compliance:
no
Remarks:
(predates GLP)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
- Name of test material: Benzenesulfonic acid, C10-13-alkyl derivs., sodium salts
- Substance type: Pure active substance
- Supplier: Japan soap and detergent association
- Analytical purity: 20.5%
- Composition of test material, percentage of components: Quaternary: 28.05%, Total amine: 0.2%,
Water: 78.7%

Test animals

Species:
monkey
Strain:
other: Macaca mulatta
Details on species / strain selection:
Macaca mulatta rhesus monkeys
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: 18-36 months
- Weight at study initiation: 2.0-4.4 Kg
- Fasting period before study: Not specified
- Housing: Animals were housed individually in wall-mounted cages
- Diet: Food consisted of 300 g dry diet and a slice of whole meal bread daily; fresh fruit was offered on alternate days. Vitamin B-12 and ascorbic acid supplements were added to the drinking water once weekly.
- Water: Tap water; ad libitum
- Acclimation period: Not specified
ENVIRONMENTAL CONDITIONS
- Temperature: 22±1°C
- Photoperiod: Normal daylight was allowed

Administration / exposure

Route of administration:
other: Test substance was administered by simultaneous oral gavage and subcutaneous injection (into the dorsal aspect of the upper thorax)
Vehicle:
water
Details on exposure:
- PREPARATION OF DOSING SOLUTIONS:
Test substance was dissolved in distilled water corresponding to doses of 0, 30, 150, 300 mg/kg bw/day for oral administration and 0.1, 0.5 and 1.0 mg/kg bw/day for subcutaneous administration.
Dose volume: 4 mL/kg (oral route) and 0.17 mL/kg (subcutaneous route)
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
28 days
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
oral route
Dose / conc.:
30 mg/kg bw/day (nominal)
Remarks:
oral route
Dose / conc.:
150 mg/kg bw/day (nominal)
Remarks:
oral route
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
oral route
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
subcutaneous route
Dose / conc.:
0.1 mg/kg bw/day (nominal)
Remarks:
subcutaneous route
Dose / conc.:
0.5 mg/kg bw/day (nominal)
Remarks:
subcutaneous route
Dose / conc.:
1 mg/kg bw/day (nominal)
Remarks:
subcutaneous route
No. of animals per sex per dose:
3 males and 3 females per dosing group
Control animals:
yes, concurrent vehicle

Examinations

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: once weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY: No

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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Before dosing commenced and again during the final week of administration, it was performed.
- Dose groups that were examined: All groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Before dosing commenced and again during the final week of administration, it was performed.
- Anesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: All animals
- Parameters: Erythrocyte sedimentation rate, packed cell volume, hemoglobin, red
blood cell count, reticulocytes, mean cell volume, mean corpuscular hemoglobin count, platelets, prothrombin index, white blood cell count total and differential.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Before dosing commenced and again during the final week of administration, it was performed.
- Anesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: all animals
- Parameters: Plasma urea and glucose, serum proteins, alkaline phosphatase, glutamic pyruvic transaminase, leucine aminopeptidase, bilirubin, sodium and potassium.

URINALYSIS: Yes
- Time schedule for collection of urine: Before dosing commenced and again during the final week of administration, it was performed.
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Not specified
- Parameters examined from 20-hours samples, after centrifugation were specific gravity, pH, protein, reducing substances, glucose, ketones, bile pigments, urobilinogen and hemoglobin. Deposits were examined for the presence of cells, casts, organisms or other abnormal constituents.

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
At termination, animals were euthanized under deep sodium pentobarbitone anesthesia and autopsied, the brain, pituitary, heart, lungs, liver, spleen, pancreas, thymus, prostate/uterus, kidneys, thyroids, adrenals and gonads were weighed. Representative samples of all body tissues including the injection sites were preserved by the appropriate method, subsequently sectioned and stained for histopathological examination

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
All monkeys receiving 300 p.o. mg/kg bw/day vomited on numerous occasions, usually within 3 hours of being dosed. Vomiting was often associated with salivation and/or retching. There was also a marked increase in the frequency of passage of loose or liquid faeces at 150 p.o. and 300 p.o. mg/kg bw/day.
Mortality:
not specified
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was no significant body weight gain, although individual animals from the group given 300 mg oral/kg/day showed slight depression of body weight gain.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
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 specified
Details on results:
CLINICAL SIGNS: All monkeys receiving 300 p.o. mg/kg bw/day vomited on numerous occasions, usually within 3 hours of being dosed. Vomiting was often associated with salivation and/or retching. There was also a marked increase in the frequency of passage of loose or liquid faeces at 150 p.o. and 300 p.o. mg/kg bw/day.

BODY WEIGHT AND WEIGHT CHANGES: There was no significant body weight gain, although individual animals from the group given 300 mg oral/kg/day showed slight depression of body weight gain.

OTHER EXAMINATION:
There was a dose-related increase in the occurrence of chronic inflammatory cell infiltration {mainly fibroblasts) at the subcutaneous injection sites. These changes were associated with the presence of pseudocysts, haemorrhage and necrosis in the dosed animal.

There was no adverse effect on food and water consumption. There was no further evidence of treatment related responses on the ophthalmological findings, hematogical findings, clinical chemistry, gross pathology and histopathology findings.

Effect levels

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
(oral)
Effect level:
150 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
food consumption and compound intake
water consumption and compound intake
ophthalmological examination
haematology
clinical biochemistry
urinalysis
Remarks on result:
other:
Remarks:
An increased frequency of loose or liquid faeces was recorded for animals receiving 150 (po) and 0.5 (sc) mg/kg bw/day. These effects were probably related to the inherent irritative effects of LAS rather than to its systemic toxicity. All monkeys receiving 300 p.o. mg/kg bw/day vomited on numerous occasions, usually within 3 hours of being dosed and may not have been truly exposed to LAS. Therefore, NOAEL (oral; systemic) was found to be 150 mg/kg bw/day (nominal)
Key result
Dose descriptor:
NOAEL
Remarks:
(subcutaneous)
Effect level:
0.5 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Remarks on result:
other:
Remarks:
An increased frequency of loose or liquid faeces was recorded for animals receiving 150 (po) and 0.5 (sc) mg/kg bw/day. These effects were probably related to the inherent irritative effects of LAS rather than to its systemic toxicity. All monkeys receiving 300 p.o. mg/kg bw/day vomited on numerous occasions, usually within 3 hours of being dosed and may not have been truly exposed to LAS. Therefore, NOAEL (oral; sc) was found to be 0.5 mg/kg bw/day (nominal)

Any other information on results incl. tables

Table1: Responses to simultaneous oral and subcutaneous administration (Heywood et al., 1978)

Dose (mg a.i./kg/day) LAS Observations during treatment period
Number of monkeys Number of occurrences
Vomiting Passing abnormal Vomiting Passing abnormal
faeces faeces
Controls 2 2 13 12
30 p.o. and 0.1 s.c. 2 3 3 6
150 p.o. and 0.5 s.c. 6 6 19 63
300 p.o. and 1 s.c. 6 6 67 65
Dose (mg a.i./kg/day) LAS Observations during treatment period
Number of monkeys Number of occurrences
Vomiting Passing abnormal Vomiting Passing abnormal
faeces faeces
Controls 2 2 13 12
30 p.o. and 0.1 s.c. 2 3 3 6
150 p.o. and 0.5 s.c. 6 6 19 63
300 p.o. and 1 s.c. 6 6 67 65

Table 2: Dose related increase occurrence of chronic inflammatory cell infiltration {mainly fibroblasts) in animals of each group. (Heywood et al., 1978)

Controls 30 p.o. and 0.1 s.c. 150 p.o. and 0.5 s.c. 300 p.o. and 1 s.c.
0/6 3/6 6-May 6/6

Applicant's summary and conclusion

Conclusions:
Administration of C10-13 sodium linear alkylbenzene sulphonate to monkeys at dose levels of 0, 30, 150 and 300 mg/kg/day by oral route and 0, 0.1, 0.5 and 1.0 mg/kg bw/day resulted in a systemic NOAEL of 150 mg/kg bw/day (po) + 0.5 mg/kg bw/day (sc). An increased frequency of loose or liquid faeces for animals receiving 150 and 300 mg/kg bw/day (po) and 0.5 and 1.0 mg/kg bw/day (sc) indicated that these effects are probably related to the inherent irritative effects of LAS rather than to its systemic toxicity. Animals vomited at the highest dose level and may not have been truly exposed to LAS.
Executive summary:

A 28-day repeated dose toxicity was conducted with C10-13 LAS, 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, systemic toxicity NOAELs of 150 mg/kg bw/day (oral) and 0.5 mg/kg bw/day (sc) were established (Heywood, 1978).