Glycine, N-coco acyl derivs., sodium salts

Endpoint summary

Description of key information

The acute oral toxicity of the test item was examined in ICR mice in a study according to OECD Guideline 425. In this study no male or female mouse died following single oral administration of the test item at dose levels of 1000 or 2000 mg/kg bw. Loss of vigor was observed in one case in the male 2.0 g/kg group, wet hair around the anus was observed in one case in each of the male 1.0 g/kg, male 2.0 g/kg and female 2.0 g/kg groups during the observation period for acute toxicity symptoms. No abnormality was observed six hours following administration and onwards. Therefore it is considered that these symptoms are temporary and mild. Body weight increase was inhibited one day following administration in the male 1.0 g/kg group. This was not observed in the male 2.0 g/kg group. In autopsies, a cyst in the left kidney in one case and edema in the uterus in one case were observed in the female 1.0 g/kg group. Edema in the left ovary was observed in one case in the female 2.0 g/kg group. However, these conditions develop spontaneously in this type of mouse, and it is considered irrelevant to the administration of the test article. Therefore, from this study an oral LD50 value of > 2000 mg/kg bw can be derived.

The acute dermal toxicity of the test item “Hostapon SG dried” was investigated in an OECD 402 study in accordance with the principles of GLP. Five male and five female Wistar rats were treated with the test substance at 2000 mg/kg by dermal application. The test item was formulated in polyethylene glycol 300 as vehicle. The application period was 24 hours. Following a 14 day observation period all animals were necropsied and examined macroscopically. No intercurrent deaths occurred and no clinical signs were recorded throughout the entire observation period. The only substance related effects consisted of very slight to slight dermal irritation reactions in form of erythema and/or oedema in one male and two females during the observation period. No macroscopic findings were observed at necropsy. Based on the study results the median lethal dose (LD50) of the test item after single dermal administration to rats of both sexes was greater than 2000 mg/kg body weight.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

GLP compliance:

not specified

Test type:

up-and-down procedure

Limit test:

no

Specific details on test material used for the study:

Name: Sodium cocoyl glycinate
Lot No.: 970925
Description: Clear liquid
Composition: 31.5% solution
Storage: Cool, dark place
Supplier: Fine Chemicals Development Laboratory, Central Research Laboratories, Ajinomoto Co., Inc.

Species:

mouse

Strain:

ICR

Sex:

male/female

Details on test animals or test system and environmental conditions:

4-week-old male and female ICR mice were purchased from Charles River Japan. After the 7-day quarantine and acclimatization period, 15 male and 15 female mice that did not have any abnormality in their general condition or growth and whose body weights were similar to the mean body weight were selected for use in the study. The animals were stratified based on body weight, and then randomly allocated to groups (5 mice of each sex per group). The age of the animals at the time of administration was 5 weeks old, and the body weight of the males was 22.8 to 24.8 g, and the body weight of the females was 19.7 g to 23.8 g. This type of animal was chosen because it is most commonly used in acute toxicity studies and there are much background data.
The animals were kept in a polycarbonate cage in an animal-keeping room. The temperature was set at 23 ± 3°C, humidity was set at 55 ± 15%, ventilation was set at 12 cycles per hour (all-fresh-air method) and the lighting period was 12 hours. Three to five mice were kept in one cage during the acclimatization period, and five mice were kept in one cage during the study period. White flakes were laid on the bottom of the cage and were replaced with new flakes once a week. Food was withheld for the period beginning on the afternoon of the day before administration and until 2 hours following administration. At all other times the animals had free access to solid food for laboratory animals CRF-1 and tap water, both during the acclimatization and study periods.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Distilled water for injection was added to 1.6 g or 3.2 g of the test item and mixed. Distilled water for injection was further added to the mixture to prepare 20 mL of 2.5% or 5% (w/v) test item solutions. The solutions were stored in a refrigerator until required for use.
In accordance with Guidelines for Safety Studies in Guidelines for Safety Evaluation of New Cosmetics, two doses were established in this study, i.e. 2000 or 1000 mg/kg bw. The test solution was orally administered forcedly to mice using a flexible Teflon tube. Food was withheld for 18 hours prior to administration. 0.4 mL per 10 g of body weight was administered once during the morning of the day of administration. The mice in the negative control group received distilled water for injection in the same manner as mice in the test article group.

Doses:

2000 or 1000 mg/kg bw

No. of animals per sex per dose:

5 / dose / sex

Control animals:

yes

Details on study design:

Acute toxicity symptoms were observed for two hours immediately following administration. General condition was observed during the afternoon of the day of administration (six hours following administration), and each morning until 14 days following administration (once a day).
All animals were weighed at the time of administration, 1, 7 and 14 days following administration.
All animals were sacrificed humanely by bleeding under ether anesthesia 14 days following administration. Organs were macroscopically observed.

Statistics:

Bartlett’s test for homogeneity was conducted for body weight distribution. If the distribution in both groups was equivalent, one-way analysis of variance (ANOVA) was carried out. If ANOVA revealed a significant difference between the groups, Dunnett’s multiple comparison test (a paired comparison of the mean values of the control and the test article groups) was performed. If the distribution was not equivalent, a Kruskal-Wallis rank-test was performed. If the rank-test revealed a significant difference between the groups, Dunnett’s multiple comparison test (a paired comparison of the mean values of the control and the test article groups) was performed

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No mortality was observed during the study in any group.

Clinical signs:

other: Loss of vigor was observed in one case in the male 2.0 g/kg group, wet hair around the anus was observed in one case in each of the male 1.0 g/kg, male 2.0 g/kg and female 2.0 g/kg groups during the observation period for acute toxicity symptoms. No abnor

Gross pathology:

A cyst in the left kidney in one case and edema in the uterus in one case were observed in the female 1.0 g/kg group. Edema in the left ovary was observed in one case in the female 2.0 g/kg group.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

From this study an oral LD50 value of > 2000 mg/kg bw can be derived.

Executive summary:

In this study no male or female mouse died following single oral administration of the test item at dose levels of 1000 or 2000 mg/kg bw. Loss of vigor was observed in one case in the male 2.0 g/kg group, wet hair around the anus was observed in one case in each of the male 1.0 g/kg, male 2.0 g/kg and female 2.0 g/kg groups during the observation period for acute toxicity symptoms. No abnormality was observed six hours following administration and onwards. Therefore it is considered that these symptoms are temporary and mild. Body weight increase was inhibited one day following administration in the male 1.0 g/kg group. This was not observed in the male 2.0 g/kg group. In autopsies, a cyst in the left kidney in one case and edema in the uterus in one case were observed in the female 1.0 g/kg group. Edema in the left ovary was observed in one case in the female 2.0 g/kg group. However, these conditions develop spontaneously in this type of mouse, and it is considered irrelevant to the administration of the test article. Therefore, from this study an oral LD50 value of > 2000 mg/kg bw can be derived.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-08-24 to 2010-11-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test Animals:
Animals: Rat, RccHan: WIST(SPF)
Rationale: Recognized by international guidelines as a recommended test system.
Breeder: Harlan Laboratories B.V., Kreuzelweg 53, 5961 NM / The Netherlands
Number of Animals per Group: 5 males and 5 females
Total Number of Animals: 5 males and 5 females
Age (when treated): Males: 9 weeks, Females: 11 weeks
Body Weight Range (when treated): 228 g – 253 g (males), 189 g – 207 g (females)
Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked during acclimatization.
Randomization: Selected by hand at time of delivery. No computer generated randomization program.
Acclimatization: Eight days under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

Environmental Conditions:
Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 22 ± 3 °C and for relative humidity between 30-70%, automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.
Accommodation: During acclimatization in groups of five per sex in Makrolon type-4 cages with standard softwood bedding. Individually in Makrolon type-3 cages with standard softwood bedding (‘Lignocel’ J. Rettenmaier&Söhne GmbH&CoKG, 73494 Rosenberg / Germany, imported by Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) during treatment and observation. Paper enrichment, Reference no. 207057, batch no. 67, (Enviro-dri from Lillico, Biotechnology, Surrey / UK) was included.
Diet: Pelleted standard Teklad Rat-Mouse Diet 2914C, irradiated, batch no. 30/10 (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) ad libitum. Results of analyses for contaminants are archived at Harlan Laboratories Ltd.
Water: Community tap water from Füllinsdorf ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at Harlan Laboratories Ltd.

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Details on dermal exposure:

Preparation of Dose Formulations:
Dose levels are in terms of the test item as supplied by the Sponsor. The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle added (weight:volume). The formulation was prepared shortly before the application using a magnetic stirrer, a spatula and an Ultra-Turrax as homogenizers. Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

Test Item Administration:
One day before treatment, the backs of the animals were clipped with an electric clipper, exposing an area of approximately 10% of the total body surface. Only those animals without injury or irritation on the skin were used in the test. On test day 1, the test item was applied evenly on the intact skin with a syringe and covered with a surgical gauze pad (ca. 5 x 5 cm) held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and an elastic adhesive restrainer bandage wrapped around the abdomen.
The application volume was 6 mL/kg.
Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and drapped off with disposable paper towels. Thereafter, the reaction sites were assessed. All animals were re-shaved on test day 8 to facilitate the reading of the local reactions.
Rationale: Dermal administration was used as this is one possible route of human exposure during manufacture, handling and use of the test item.

Duration of exposure:

24 hours

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

The purpose of this study was to assess the acute dermal toxicity of Hostapon SG dried when administered to rats by single semi-occlusive dermal application, followed by an observation period of 14 days. This study should provide a rational basis for risk assessment.

Vehicle:
The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. Therefore, solubility testing was excluded from the statement of compliance. According to the Sponsor, the test item is soluble in water up to a concentration of 25%. Therefore, polyethylene glycol 300 (PEG 300) was used as vehicle and the test item could be dispersed at the technically highest concentration of 33 % (w/w). This was achieved with an Ultra Turrax and resulted in a white liquid, which was considered dermally applicable.
The following information was provided by Harlan Laboratories Ltd.:
Identification: Polyethylene glycol 300 (PEG 300)
Description: Colorless viscous liquid
Lot Number: STBB 3451
Source: Sigma-Aldrich Chemie GmbH, Riedstr. 2, D-89555 / Germany
Stability of the Vehicle: Stable under storage conditions
Expiry Date: 30-Apr-2012
Storage Conditions: At room temperature (range of 20 ± 5 °C), light protected.
Safety Precautions: Routine hygienic procedures were used to ensure the health and safety of the personnel.

Observations:
Viability / Mortality: Daily during the acclimatization period. Once before treatment, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15.
Clinical Signs: Daily during the acclimatization period. Once before treatment, within the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15. All abnormalities will be recorded.
Local Dermal Signs: Once daily during days 2 (following dressing removal) through day 15 using the numerical scoring system described in Appendix I.
Body Weights: On test days 1 (prior to administration), 8 and 15.

Pathology:
Necropsy: All animals were sacrificed at the end of the observation period by carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. An external examination and opening of the abdominal and thoracic cavities for examinations of major organs was performed. The appearance of any macroscopic abnormalities was recorded. No organs or tissues were retained.

Data Compilation:
Viability/mortality was recorded on data sheets.
Body weights were recorded on-line with the ToxControl Computer System.
Clinical signs, local dermal signs, mortality data and macroscopic findings were compiled into the ToxControl Computer System during recording.
The ToxControl Computer System has been licenced for Harlan Laboratories Ltd. and validated with respect to data collection, storage and retrievability.

Statistics:

No statistical analysis was used.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No intercurrent deaths occurred during the course of the study.

Clinical signs:

other: No clinical signs were recorded throughout the entire observation period.

Gross pathology:

No macroscopic findings were recorded at necropsy.

Local dermal signs:

Very slight to slight erythema was observed on test day 2 (1 male) or from test day 2 to test day 4 (1 female) or test day 6 (1 male). Very slight to slight oedema and slightly maculated crusts were observed in one female (No. 10) from test day 2 to 3 or from test day 7 to 15, the end of the observation period. Slight desquamation was noted in one male and three females during test day 3 to test day 7 or 11 and in one male from test day 8 to test day 12. Slight to moderate desquamation was noted in one female from test day 3 to test day 14. The test item caused white to yellow discoloration of the treated skin in two males and one female on test day 2.

Interpretation of results:

not classified

Remarks:

Migrated information

Conclusions:

The median lethal dose of Hostapon SG dried after single dermal administration to rats of both sexes, observed over a period of 14 days, is:
LD50 (rat): greater than 2000 mg/kg body weight

Executive summary:

The acute dermal toxicity of `Hostapon SG dried` was investigated in an OECD 402 study in accordance with the principles of GLP. Five male and five female Wistar rats were treated with the test substance at 2000 mg/kg by dermal application. The test item was formulated in polyethylene glycol 300 as vehicle. The application period was 24 hours. Following a 14 day observation period all animals were necropsied and examined macroscopically. No intercurrent deaths occurred and no clinical signs were recorded throughout the entire observation period. The only substance related effects consisted of very slight to slight dermal irritation reactions in form of erythema and/or oedema in one male and two females during the observation period. No macroscopic findings were observed at necropsy. Based on the study results the median lethal dose (LD50) of Hostapon SG dried after single dermal administration to rats of both sexes was greater than 2000 mg/kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

In both studies with oral and dermal dosing, LD50 values of > 2000 mg/kg were obtained. Therefore, there is no need for classification and labelling of the test item according to CLP Regulation 1272/2008/EG.