Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1975
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study was performed before GLP principles were in place. Several study limitations compared to OECD 414: no analytical verification of doses performed, limited monitoring of health parameters of the parents; rats not weighed during study, only at sacrifice; organ weight of parents and/or offspring was not included; only two dose levels included (instead of≥3); sacrifice for teratological examination at day 13 (10/20 rats) or day 20 (10/20 rats). Test substance (Aerosol A-268) is a formulation with appr. 47.5% disodium 4-isodecyl sulfosuccinate, thus it cannot be excluded that the effects seen are related to formulation toxicity.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1975
Report Date:
1975

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
performed in 1975
Deviations:
yes
Remarks:
Only two dose levels tested. No chemical analyses of test substance. Limited clinical observations, only weight at sacrifice. 10/20 dams killed at GD13, 10/20 at GD20.
Principles of method if other than guideline:
Weaning rats (male and female) were exposed via feed. At 15 weeks, they were paired and offspring was monitored (teratoloy/ histopathology). In parallel, rats of the F1 generation were allowed to grow up and mate, litters were examined (teratoloy/ histopathology). P and F1 generation parents, and weanling rats of F1 and F2 were examined histopathologically.
GLP compliance:
no
Remarks:
Study performed before GLP principles were in place
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): AEROSOL® A-268
- Analytical purity: 50% disodium 4-isodecyl sulfosuccinate formulation in water

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River
- Age at study initiation: (P) 3 wks
- Fasting period before study: no
- Housing: individually, in suspended steel cages
- Diet: ad libitum, Charles River 19RF meal
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Controlled environmental room
- Temperature (°C): appr. 23
- Humidity (%): 45-65
- Photoperiod (hrs dark / hrs light): appr. 12/12

- No details on in-life data

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
- A-268 was thoroughly blend in the feed at 1% or 4% (no details on frequency diet preparation).
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
When the animals (P1 or F1) were sexually mature (appr. 15 weeks), one male was placed with one female in mating cages. The males within each group were rotated every sixth day and removed on the 19th day. The females were allowed 20 days rest between weaning and the next breeding.
Pregnancies for the teratology phase were timed by daily vaginal smear of by vaginal plug visualization.
Duration of treatment / exposure:
P1: start at weaning (appr. 3 weeks of each) until sacrifice (during pregnancy of third litter), estimated minimal exposure time is 33 weeks;
F1: exposure in utero, from day of birth via mother milk and after weaning through feed, estimated minimal exposure time is 36 weeks;
F2: 4 days (exposure in utero and via milk)
Frequency of treatment:
Continuous (through feed)
Duration of test:
- P generation was exposed from weaning and mated at 15 weeks. They were allowed to have three litters: First litter was sacrificed at weaning and from the second litter parental animals of the F1 generation were selected. The fetuses from the third litter were used to investigate teratological effects.
- F1 parental animals not mated until appr. 12 weeks after selection from the F1 litters. First litter was sacrificed at weaning, the second litter was used to investigate teratological effects.
No. of animals per sex per dose:
20
Control animals:
yes, plain diet
Details on study design:
Summary of generations
P1 = Parental generation, allowed 3 litters
 
F1a = discarded at weaning
F1b = allowed 2 litters
F1c = Teratological examination and histology (5/sex/ group)
 
F2a = discarded at weaning
F2b = Teratological examination and histology (5/sex/ group)

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: only at sacrifice

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 13 or 20
- Organs from five dams from each treatment group were evaluated histologically during teratological phase (organs include (but are not limited to) thyroids, lung, heart, thymus, spleen, liver, kidneys, uterus, ovaries and brain). In case any abnormalities were observed, then the testes from the corresponding fathers (5/ group) were also evaluated histologically.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Half of the dams was sacrificed at GD 13, the other half at GD 20.
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: Ovaries of 5 dams were evaluated by histopathology.
Fetal examinations:
Fetuses (collected from dams sacrificed at GD 20) were inspected for gross abnormalities, sexed and litter-weighed. One third of the fetuses were cleared, stained with alizarin red stain and examined for skeletal abnormalities. The remaining fetuses were examined for soft tissue abnormalities either by histological methods, or by freehand sectioning.
Statistics:
Statistical analysis of litter sizes were performed with analysis of variance.
Indices:
Fertility Index = (number of pregnancies/ number of matings) x 100;
Viability Index = (number of live pups at four days/ number of live pups born) x 100;
Lactation Index = (number of weaned pups/ (number of live pups-number of pups discarded at day 4))x100.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
No substance-related mortality was found and no substance-related histopathological effects were seen.
The fertility index was decreased for the highest concentration group calculated for both litters of the P generation (95%, 95%, 70% and 95%, 90% and 80% for the control group and 1% and 2% diet concentration for the first and second litter resp.). For the first litter of the second generation the fertility index of the three groups did not differ (all 95%).

Effect levels (maternal animals)

open allclose all
Dose descriptor:
LOAEL
Effect level:
1 other: %
Based on:
test mat.
Remarks:
(conc. in feed based on solid)
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOAEL
Effect level:
>= 4 other: %
Based on:
test mat.
Remarks:
(conc. in feed based on solid)
Basis for effect level:
other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
Both at day 13 and day 20 of pregnancy the average number of viable embryos per litter was decreased in a concentration-dependent manner for embryos of the P and the F1 generation. The number of early and/or late resorptions did not differ between the exposed and non-exposed groups for both generations. The number of abnormal fetuses was not increased in the rats of the P and the F generation of exposed rats at 20 days of pregnancy.

VIABILITY (OFFSPRING)
A substance-related decrease in the average number of pups/ litter was observed in all litters (11.8, 10.3, and 7.7 (F1a generation); 10.9, 10, 8.2 (F1b-generation); 11.1, 10.3 and 5.4 (F2a generation) for resp. 0%, 1% and 4% dose groups. The viability of life-born pups until weaning was decreased only at the highest dose for the first litters of both generation (viability index for resp. 0%, 1% and 4%: 99.1, 98.9 and 84.7 (F1a); 98.1, 100.0 and 98.5 (F1b); 98.1, 98.9 and 82.9 (F2a). The lactation index was decreased for the first litter of the P generation (96.5, 93.4, 69.1 for control, 1% and 4% groups). The second litter of the P generation did not show influence of the test substance (resp. 85.2, 94, 97.5 for control, 1% and 4% groups). However, also the first litter of the second generation showed an increased pup loss during the first three weeks at the highest dose level as shown by a lactation index of 96.5, 99.3, 86.1 for the control, 1% and 4% group.

BODY WEIGHT (OFFSPRING)
The average weaning weight of pups decreased in a dose-related fashion for both male and female pups. This effect was seen in the first and second litter of the P generation, and also in the first litter of the F1 generation (male pups: -17% and -58% (F1a-pups), -13% and -41% (F1b-pups) and -16% and -61% (F2a-pups) for resp. 1% and 4% dose groups compared to control group; female pups: -11% and -54% (F1a-pups), -20% and -60% (F1b-pups) and -17% and -60% (F2a-pups) for resp. 1% and 4% dose groups compared to control group).

SEXUAL MATURATION (OFFSPRING)
The fertility indices of the F1 generation were not affected by the test substance (95% for control and both dose groups).

GROSS PATHOLOGY (OFFSPRING)
A male weanling of the F1 control group had mild respiratory disease.,

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Based on the results of a reproduction/ teratology study with rats, an LOAEL of 1% in feed for reproduction and development of A-268 was found in absence of systemic toxicity of the parents. At the LOAEL, a decreased number of viable embryos at day 13 and 20 of pregnancy was found, corresponding to dose-related increase of number of implantation sites. Furthermore, pups showed reduced weight (>10%) at weaning, and a dose-related increase of mortality of pups between day 4 and weaning was seen at 1% and higher. 1% corresponds to appr. 500 mg solids/kg bw/day exposure of the mother. Test substance (AEROSOL® A-268) is a formulation with appr. 47.5% disodium 4-isodecyl sulfosuccinate, exact composition is unclear. Based on the available data, the LOAEL of disodium 4-isodecyl sulfosuccinate is estimated to be 475 mg/ kg bw/ day.
Executive summary:

A multi-generation study was performed similar to OECD 414 and 416. Male and female rats (20/sex) were exposed to A-268 via feed at 0%, 1% or 4% from weaning until reproductive age. Rats were mated several times: first litter was sacrificed at weaning, the second litter was allowed to mature and the rats were sacrificed at day 13 (10/20) or day 20 (10/20) of the third pregnancy. The first litter of the F1 generation was sacrificed at weaning, F1 rats were sacrificed at day 13 and 20 of the second pregnancy (10 rats/ timepoint). No substance-related mortality occurred in the parental animals of P and F1 generation, no substance-related histopathology was found. The fertility index was decreased for the highest concentration group calculated for both litters of the P generation, but not for the first litter of the second generation. The number of viable fetuses (average/ litter) was decreased in a dose-related manner for all litters at both concentrations. This corresponds to a dose-related decrease in the number of implantation sites in both exposed groups seen in rats sacrificed at day 13 of pregnancy. No substance-related anomalies were found in skeletal or soft tissue of foetuses at day 20 of pregnancy. The viability of the pups until weaning was decreased in the first litters of the P and F1 generation at the highest dose only. The average weaning weight of pups was decreased (at least with 10%) at both dose levels in all litters. Based on these results, an LOAEL for development of 1% A-268 was established (corresponding to a dose level of appr. 500 mg solids/kg bw/ day) in absence of systemic toxicity of the parents.

Based on these results, an LOAEL for reproduction and development of 1% A-268 was established (corresponding to a dose level of appr. 500 mg/kg bw/ day) in absence of systemic toxicity of the parents. Based on the available data, the LOAEL of disodium 4-isodecyl sulfosuccinate is estimated to be 475 mg/ kg bw/ day.