Reaction mass of glycerol 1,3-di(acetate) and glycerol acetate and triacetin

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP-Guideline study, tested with the source substance Triacetin (CAS No 102-76-1). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1998

Reference 2

Reference Type:

secondary source

Title:

Triacetin CAS No. 102-76-1

Author:

OECD SIDS

Year:

2002

Bibliographic source:

SIDS Initial Assessment Report For SIAM 15

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Triacetin

EC Number:

203-051-9

EC Name:

Triacetin

Cas Number:

102-76-1

Molecular formula:

C9H14O6

IUPAC Name:

propane-1,2,3-triyl triacetate

Details on test material:

- Name of test material (as cited in study report): Triacetin
- Physical state: colourless clear liquid with slight odour
- Source: Daihachi Chemical Industry. Co., Ltd.
- Analytical purity: > 98.2 %
- Lot/batch No.: N-80302
- Stability under test conditions: Stability confirmed during use by gas chromatography.

Test animals

Species:

rat

Strain:

other: Crj: CD(SD) IGS

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: (P) 9 weeks
- Weight at study initiation: (P) Males: 371-375 g; Females: 203-240 g

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 3 % gum arabic in purified water

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: up to 7 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear

Duration of treatment / exposure:

44 days (males) from 2 weeks prior to mating.
41-48 days (females) from 14 days before mating to day 3 postpartum.

Frequency of treatment:

daily

Duration of test:

Day 14 before mating to Day 3 postpartum

Doses / concentrations

Remarks:

Doses / Concentrations:
40, 200 and 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

12

Control animals:

yes, concurrent vehicle

Examinations

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: The general condition was observed once a day.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weight was determined on Day 0, 3, 7 and 14 of administration and once a week thereafter. For pregnant females, body weight was determined on the Day 0, 14 and 20 of gestation and on Day 0 and 4 of lactation.

FOOD CONSUMPTION: Yes
Food consumption was determined on the same day when body weight was measured.

HISTOPATHOLOGY / ORGAN WEIGHTS
Microscopic: all animals in control and 1,000 mg/kg bw group, and unfertilized animals in other groups: brain, spinal cord, pituitary gland, eyeball, thyroid gland (including parathyroid gland), thymus, heart, trachea, lung, liver, kidney, adrenal, spleen, stomach, small intestine, large intestine, pancreas, urinary bladder, bone marrow, sciatic nerve, lymph node, testes, epididymis, prostate, seminal vesicle, ovary, uterus, vagina, mammary gland and any organs, which might be expected to have histopathological changes and thymus and lung of dead animals.
Organ weight: for both sexes, brain, pituitary gland, thyroid gland, heart, liver, kidney, spleen, adrenal, thymus, and in addition for males, testes and epididymis.

OTHER:
Haematology and biochemistry for males conducted only at time of necropsy after 44 days of exposure (for further details refer to 7.5.1 Repeated dose toxicity).

Ovaries and uterine content:

- Number of corpora lutea: Yes
- Number of implantations: Yes

Fetal examinations:

Numbers of offspring or live offspring, the sex ratio, the live birth index, the viability index and body weights, external features, clinical signs and necropsy.
Gross necropsy consisted of external and internal examination.

Statistics:

Regarding quantitative data (body weight, gain of body weight, food consumption, organ weight, haematology, clinical chemistry, number of corpora lutea, number of implantation sites and total number of offspring), Bartlett test was used. In case of equal variance and unequal variance, ANOVA and Kruskal-Wallis test was applied, respectively. If there was a significant difference, Dunnett test or Dunnett multiple-comparison was used. For day of conceiving, number of estrus, gestation length, implantation index, delivery index, viability index at Day 0 and Day 4, Bartlett test and Kruskal-Wallis test was used. If a significant difference was found, Dunnett multi-comparison test was applied. For histopathology findings, Chi-square was used. If a significant difference was observed, Chi-square of Armitage test was used between control and administration group. Regarding copulation index, fertility index, gestation index and sex ratio of offspring was tested with Fisher’s exact test.

Indices:

Copulation index (%): Number of copulated females/number of pairs x 100
Fertility Index (%): Number of pregnant females/numer of copulated females x 100
Gestation index (%): Number of pregnant animals delivered live offspring/ numer of pregnant animals x 100
Delivery index (%) and implantation index (%) were given.
The gestation index, gestation length, parturition and maternal behavior. Effects at gross pathology and microscopic evaluation of the reproductive organs.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No dose-related changes in general clinical signs. One male at 1000 mg/kg bw/day was dead 32 days after the administration started.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
For both sexes, no statistically significant difference from controls was observed in body weight and body weight gain during administration period. For both sexes, no statistically significant difference from controls was observed in food consumption during administration period.

ORGAN WEIGHTS (PARENTAL ANIMALS)
No dose-related changes in organ weight were observed.

GROSS PATHOLOGY (PARENTAL ANIMALS)
No changes in gross pathology in both sexes were observed.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Tissue pathology revealed no alteration of tissues even in the highest dose groups for both sexes.

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
On examination of neonates, there were no significant differences in numbers of offspring or live offspring, the sex ratio, the live birth index, the viability index or body weights. No abnormal findings ascribable to the compound were found for external features, clinical signs or necropsy of the offspring. One dead offspring showed pyelectasis at dosing of 40 mg/kg bw/day.

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Table 1. Delivery data (P).

Parameter [mean]	Group 1 0 mg/kg bw	Group 2 40 mg/kg bw	Group 3 200 mg/kg bw	Group 4 1000 mg/kg bw
Number of corpora lutea	16.6	16.3	16.3	16.8
Number of Implantations sites	15.3	16.0	14.7	15.8
Total number of offsping	14.4	15.3	14.3	14.6
Implantation Index (%)	90.15	98.17	89.49	93.84
Delivery Index (%)	94.83	95.11	90.17	92.62
Gestation Index (%)	100	100	91.7	100

Table 2. Litter size and viability index (F1).

Parameter	Group 1 0 mg/kg bw	Group 2 40 mg/kg bw	Group 3 200 mg/kg bw	Group 4 1000 mg/kg bw
Total number of offspring at birth	14.4	15.3	15.6	14.6
Total number of live offspring at birth	14.2	15.3	15.5	14.6
Number of live offspring on Day 4	14.1	14.8	15.2	14.5
Viability index (%) Day 0 Day 4	98.71 99.38	100 97.17	98.86 98.3	100 99.41

Applicant's summary and conclusion

Conclusions:

The test substance had no effect on intrauterine development.