Bis(C12-C13)alkyl-2-hydroxybutandioate

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April - May 1993

Reliability:

2 (reliable with restrictions)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method B.9 (Repeated Dose (28 Days) Toxicity (Dermal))

GLP compliance:

yes

Limit test:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): Cosmacol EMI
- Substance type: pure active substance
- Physical state: liquid

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: conelli - Arona VR, Italy
- Age at study initiation: no data
- Weight at study initiation: 2.0 - 2.9 kg
- Fasting period before study: no
- Housing: individually caged in stainless stell dages (dimensions 450x500x380 mm)
- Diet (e.g. ad libitum): standard pellet complete diet ad libitum
- Water (e.g. ad libitum): filtered tap water ad libitium
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
The housing room condition of temperature and humidity were maintained by the conditioning plant and continuously recorded, no further details mentioned
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: April 05, 1993 To: May 19, 1993

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on exposure:

TEST SITE
- Area of exposure: dorsal
- % coverage: not mentioned
- Type of wrap if used: the sample was put on a gauze patch and applied on the shaved dorsal area by means of a occlusive bandage, no further details mentioned
- Time intervals for shavings or clipplings: weekly

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mL/kg bw
- Constant volume or concentration used: yes

USE OF RESTRAINERS FOR PREVENTING INGESTION: not mentioned

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

28 days

Frequency of treatment:

Duration of exposure per day: 6 hours
Dosing regime: 5 days/week

Doses / concentrations

Remarks:

Doses / Concentrations:
10 mL/kg
Basis:
nominal per unit body weight

No. of animals per sex per dose:

5

Control animals:

other: treatment with sesame seed oil

Details on study design:

- Dose selection rationale: no data
- Rationale for selecting satellite groups: no data
- Post-exposure recovery period in satellite groups: 14 days

Examinations

Observations and examinations performed and frequency:

OBSERVATIONS DURING TREATMENT:
- Mortality: every morning and every afternoon
- Clinical observations: daily, including organic body functions, tegumentary apparatus, mucosae conditions, respiratory activity, sensorium conditions, specific examination for skin irritancy carried out on the area of application
- Body weight: animals were weighed before every treatment, the body weights were recorded in the week in which the treatment started, then weekly
- Food data: the quantity of food consumed was registered weekly

OBSERVATIONS AT THE END OF THE TREATMENT:
before sacrifice a survey of the general condition was performed on all animals

LABORATORY EXAMINATIONS:
before sacrifice, a cardiac blood taking was performed for the following examinations:
- hematological examinations, including hemoglobine, erythrocytes, leucocytes, hematocrit, medium cell values, leucocytic formula, MID (monocytes, eosinophils, basophils), granulocytes, lymphocytes, blood platelets
- hematochemical examinations, including azotemia, glycaemia, bilirubinaemia, gamma GT, GOT, GPT, total proteins, calcemia, sodium, potassium

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, the organs of the two cavities were observed globally, removed and dissected individually in order to detect abnormalities.
The following organs were weighed and a slice of each and also of skin was put in 10% formaline: lungs, heart liver, spleen, kidney, suprarenal gland, gonads
HISTOPATHOLOGY: No

Statistics:

The significance of the differences between the average body weights, food consumption, haemotology data and hematochemical data was evaluated by means of Students t-test at a value of 5%.
The organ weights in relation to body weight were evaluated by means of Mann-Whitney test ranks.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Dermal irritation:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No deaths occurred during the study in any of the 3 groups (treated - satellite - control).
No clinical signs were observed during the experimental observation period in all 3 groups. In particular no erythema and/or oedema was observed in the test group.

BODY WEIGHT AND WEIGHT GAIN
Body weight gain was similar in all 3 groups.

FOOD CONSUMPTION
Food consumption was similar in all 3 groups.

HAEMATOLOGY
No significant differences in all 3 groups.

CLINICAL CHEMISTRY
Significant differences were measured in the sodium, calcium, glycaemia, azotemia and total protein in the females of the satellite group and of calcium in the males. However, these levels were within the range for the species and considered as not relevant.

ORGAN WEIGHTS
The weight of organs did no show any significant variations in relation to the treatment.

GROSS PATHOLOGY
At the ante-mortem examination in all animals no pathological symptoms were observed.
No macroscopic abnormalities related to the treatment were seen at necropsy.
Minor anomalies were observed in 2/5 males and 2/5 females of the test group, 2/5 females in the control group and 2/5 males and 2/5 females in the satellite group. These minor findings were gastric meteorism, mucosae enteritis, presence of connective laciniae on stomach wall, presence of granular tissue on stomach wall, liverworth steatosis, kidney cyst, spleen iperplasy of white pulp and splenic stasis.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

10 other: mL/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

2

Dose descriptor:

NOEL

Effect level:

10 other: mL/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

3

Dose descriptor:

NOAEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

In experimental conditions, on the basis of a total evaluation of the results obtained, the test material Bis(C12-C13)alkyl-2-hydroxybutandioate did not cause any local or general toxicity. The NOAEL was determined to be 10 mL/kg bw/day (1000 mg/kg bw/d).

Executive summary:

The study of subacute dermal toxicity (28 days) was performed on a group of 30 rabbits (20 treated and 10 controls) according to Directive 92/69/EEC.

The test material, Bis(C12 -C13)alkyl-2 -hydroxybutandioate, was administered for 4 weeks (5 days out of 7) at daily dose of 10 mL/kg bw (1000 mg/kg bw/d) by dermal application. The test material was administered undiluted to 20 animals, while the control group (10 animals) were treated with sesame seed oil.

The animals were sacrificed at the end of treatment, on the 28th days (treated and control groups) with the exception of 10 rabbits which had been treated with the test material, they were kept under observation for another 2 weeks after the end of treatment (satellite group).

During the study animals were observed daily for possible toxicity symptoms. These observations included: evaluation of local reaction in the application aria, evaluation of organ functions, evaluation of the tegumentary apparatus conditions, evaluation of possible variations in somatomotor activity and sensorium conditions, evaluation of mucosae conditions. Animals body weight and food consumed were recorded weekly. At the end of the study a blood taking from heart in all animals was executed in order to evaluate the hematological and hematochemical parameters in the three groups (treated and controls), then an autopsy was performed.

The results obtained from the study can be summarized as follows: No case of mortality and no pathological symptoms were observed in the animals treated and in the control animals, in particular no phenomena of erythema or oedema was observed in the area where the test material was applied.

At the autopsy no pathological anomalie were observed. The weight of the organs showed no significant differences between the 3 groups. The hematological and hematochemical examinations, conducted at the end of the study did not show any significant differences between the groups.

In experimental conditions, on the basis of a total evaluation of the results obtained, the test material Bis(C12-C13)alkyl-2-hydroxybutandioate did not cause any local or general toxicity.