Registration Dossier

Administrative data

Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1974-1977
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
No guideline study and non-GLP but performed according to scientific standards at time of performance. Acceptable based on nowadays existing guidlines and standards. Well performed and documented. Reliability declaration regarding procedures and performance available.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1978
Report Date:
1978

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
Two-generation reproductive toxicity study including a segment II phase for developmental toxicity
GLP compliance:
no
Remarks:
but reliability declaration from study director included
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Hostapur SAS
- Physical state: aqueous slurry
- Analytical purity: 60%
- Composition of test material, percentage of components: 60% Hostapur SAS 93, water
- Isomers composition: n.a.
- Purity test date: 1974-09-01
- Lot/batch No.: NR T 2/112
- Expiration date of the lot/batch: 1979-10-01
- Radiochemical purity (if radiolabelling): n.a.
- Specific activity (if radiolabelling): n.a.
- Locations of the label (if radiolabelling): n.a.
- Expiration date of radiochemical substance (if radiolabelling): n.a.
- Stability under test conditions: stability and homogeneity guaranteed
- Storage condition of test material: in darkness at room temperature
- Other:

Test animals

Species:
rat
Strain:
other: CD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River U.K.
- Age at study initiation: (P) = weanling rats
- Weight at study initiation: (P) Males: 79 +/- 3g; (P) Females: 87 +/- 3 g
- Fasting period before study: no
- Housing: polypropylene cages
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 2 °C
- Humidity (%): 50 +/-20%
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12 hours

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Spratts Laboratory Diet No. 2
- Storage temperature of food: refrigerator


VEHICLE
- Justification for use and choice of vehicle (if other than water): n.a.
- Concentration in vehicle: n.a.
- Amount of vehicle (if gavage): n.a.
- Lot/batch no. (if required): n.a.
- Purity: n.a.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 60 days (three pregnancies derived from each of two generations)
- Proof of pregnancy: sperm in vaginal smear referred to as day 1 of pregnancy
- After 5 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged separately
- Any other deviations from standard protocol: Approximately 20 days after weaning of the first (A) litters the females were re-mated with different males to produce the second (B) litters. This was repeated a second time to produce the third (C) litters.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
60 days prior to mating and throughout three successive pregnancies (fertility)
Gestation days 6 to 15 only for females during organogenesis stage (development)
Frequency of treatment:
daily
Details on study schedule:
- F1 parental animals not mated until approximately 10 weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were approximately 15 days of age.
- Age at mating of the mated animals in the study: approximately 13 weeks
Doses / concentrationsopen allclose all
Dose / conc.:
0 ppm
Remarks:
Doses / Concentrations:
0, 1000, 3000, 10000 ppm
Basis:
nominal in diet
Fertility groups 2, 3, 4
Dose / conc.:
1 000 ppm
Remarks:
Doses / Concentrations:
0, 1000, 3000, 10000 ppm
Basis:
nominal in diet
Developmental groups 5, 6, 7
Dose / conc.:
3 000 ppm
Remarks:
Doses / Concentrations:
0, 1000, 3000, 10000 ppm
Basis:
Dose / conc.:
10 000 ppm
Remarks:
Doses / Concentrations:
0, 1000, 3000, 10000 ppm
Basis:
No. of animals per sex per dose:
25 per sex per group
Control animals:
yes
Details on study design:
- Dose selection rationale: Based on repeated dose toxicity study and expert judgement
- Rationale for animal assignment (if not random): random
- Other:
Positive control:
not required

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily


BODY WEIGHT: Yes
- Time schedule for examinations: weekly


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food intake per cage of five rats was recorded and the mean intake per rat calculated



OTHER: Heamtological investigations, absolute and relative organ weights, histopathological investigation of tissues
Oestrous cyclicity (parental animals):
oestrous cycles, mating performance, conception rates and pre-coital interval were examined for all three matings
Sperm parameters (parental animals):
Sperm parameters not investigated in detail
Litter observations:
STANDARDISATION OF LITTERS
All offspring, except those selected to form the F1 generation, were killed after day 21 post partum by carbon dioxide asphyxiation and examined.

PARAMETERS EXAMINED
The following parameters were examined in [F1A, F1B, F1C, F2A, F2B and F2C offspring:
- number and sex of pups,
- stillbirths,
- live births,
- postnatal mortality,
- presence of gross anomalies,
- weight gain,
-physical or behavioural abnormalities,
The speed of physical development of the offspring was assessed by the following parameters:
- pinna unfolding
- hair growth,
- tooth eruption,
- eye opening
- auditory function,
- visual function


GROSS EXAMINATION OF DEAD PUPS:
[no / yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.]
Postmortem examinations (parental animals):
SACRIFICE
- At day 13 (half of dams) or day 21 (remaining dams) of the third pregnancy all parent animals were killed and a thorough gross macroscopic examination carried out

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs / tissues were prepared for microscopic examination and weighed, respectively:
- heart, liver, kidneys, ovaries, testes, adrenals, spleen, lung, stomach, pancreas, thymus, small intestine, urinary bladder

pre-implantation loss and post-implantation loss were examined
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals were sacrificed at 21 days of age.
- These animals were subjected to postmortem macroscopic and/or microscopic examination


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations


HISTOPATHOLOGY / ORGAN WEIGTHS
The folowing organs / tissues were prepared for microscopic examination and weighed, respectively:
- heart, liver, kidneys, ovaries, testes, adrenals, spleen, lung, stomach, pancreas, thymus, small intestine, urinary bladder
Statistics:
Mann-Whitney non-parametric U test; chi-squared test incorporating Yates` correction
Reproductive indices:
Mean litter size, mean litter weight, mean offspring weight (day 1, 21, 25), mean offspring weight on day 4 post partum
Offspring viability indices:
Viability index, live birth index

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed

Details on results (P0)

In both generations (F0 and F1 parental animals) food intake, haematological parameters, absolute and relative organ weights and histopathological evaluation of tissues (heart, liver, kidneys, ovaries, testes, adrenals, spleen, lung, stomach, pancreas, thymus, small intestine, urinary bladder)
showed no adverse treatment-realted effects. A slight depression of body weight gain was observed in the F0 generation in the highest dose group. No significant inter-group variation was observed during the three subsequent pregnancies. In both parental generations (F0 and F1) oestrous
cycles, mating performance and conception rates were unaffected by treatment.

Effect levels (P0)

open allclose all
Dose descriptor:
NOEL
Effect level:
>= 1 000 ppm (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: Generation: P, F1a, F1b, F2a, F2b (migrated information)
Dose descriptor:
NOAEL
Effect level:
>= 3 000 - <= 10 000 ppm (nominal)
Sex:
female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: Generation: F1a, F2b (migrated information)
Dose descriptor:
NOAEL
Effect level:
>= 10 000 ppm (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Other effects:
no effects observed

Details on results (P1)

Details on maternal toxic effects:
Slight body weight gain depression in animals of the highest dose group (10000 ppm) (continously
treated)

Effect levels (P1)

open allclose all
Dose descriptor:
NOEL
Effect level:
>= 1 000 ppm (nominal)
Sex:
not specified
Basis for effect level:
other: See remark
Dose descriptor:
NOEL
Effect level:
>= 3 000 - <= 10 000 ppm (nominal)
Sex:
not specified
Basis for effect level:
other: See remark
Remarks on result:
other: other: other: Generation: F1a, F2b (migrated information)

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Details on results (F1)

No treatment related effects in the number of litters containing at least one viable young, the litter size at birth, or the live birth index was noted in
any of the generations. Viability index slightly depressed in F1a litters of females at highest dose level. In all other groups, the viability index was
comparable to the control group. Body weight gain was slightly depressed in the F1a, F1b, F2a and F2b litters. All other treated offspring gained
weight at similar rate to the controls.Significant inter-group variations were not observed.

Effect levels (F1)

open allclose all
Dose descriptor:
NOEL
Generation:
F1
Effect level:
>= 10 000 ppm (nominal)
Sex:
not specified
Basis for effect level:
other: teratogenicity
Dose descriptor:
NOEL
Generation:
F1
Effect level:
>= 10 000 ppm
Sex:
not specified
Basis for effect level:
other: embryotoxicity

Overall reproductive toxicity

Key result
Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The NOEL with regard to reproductive performance and/or embryotoxicity was 10000 ppm in the diet (corresponding to about 500 - 675 mg/kg
body weight per day).
Executive summary:

The influence of sec-alkane sulfonate-sodium salts SAS (60%) upon reproductive function and fertility was assessed over two generations in rats of the Charles River CD strain. For this purpose, sec-alkane sulfonate-sodium salts SAS was administered in the diet to both the F0and F1generations at levels of 1000, 3000 or 10000 ppm. Treatment was given either continuously to both sexes for 60 days prior to mating and throughout three successive pregnancies (F1A, F1B, F1C, F2A, F2B and F2C) or to females only during the organogenesis stage of three successive pregnancies. Animals were randomly selected from the F1B litters to form the second generation. In both the F0and F1generations, haematological investigations were carried out after 60 days of treatment on five males and five females in each group, prior to carbon dioxide asphyxiation and subsequent macroscopic and histopathological examination. The remaining animals were paired, within groups, on a one to one basis on three consecutive occasions. After the first two matings (F1A, F1B, F2A, F2B) the females were allowed to litter naturally and rear their young to weaning. Following the third mating (F1C, F2C) half of the dams in each group were killed on day 13 of gestation, and the remainder were killed on day 21 of gestation, to permit examination of their uterine contents. After termination of each generation, all parent animals were examined macroscopically. Five males and five females from each of the continuously treated groups, together with five females only from each of the groups treated during organogenesis, were randomly selected for histopathological evaluation. In both generations prior to mating, food intake, haematological parameters, absolute and relative organweights and histopathological evaluation of tissues showed no adverse treatment-related effects.In the F0generation, a slight but not statistical significant depression of body weight gain was observed in males treated continuously with sec-alkane sulfonate-sodium salts SAS at 10000 ppm. A similar reduction was observed in F0females treated prior to mating at 10000 ppm and statistical significance was achieved here in week 8 of treatment. During the three subsequent pregnancies, some fluctuation in body weight gain was recorded in treated females, but no significant inter-group variation was observed. In both generations, oestrous cycles, mating performance and conception rates were unaffectedby treatment. In the F0and F1generation no alterations in duration of gestation were observed. Neither generation, during the first two pregnancies (F1A, F1B, F2A, F2B), showed any treatment-related effects in the number of litters containing at least one viable young, the litter size at birth, or the live birth index. The viability index was significantly depressedin the F1A litters of females receiving 3000 or 10000 ppm continuously, and in the F2B litters of the females receiving 3000 ppm continuously. In all other groups, the viability index was comparable with that of the control group. The body weight of offspring at day 1 post partum showed no significant inter-group variations. However, the bodies weight gain of offspring from females receiving 10000 ppm continuously was depressed in the F1A, F1B, F2A and F2B litters. All other treated offspring gained weight at a similar rate to the controls. In both generations, the sex ratio at day 4 post partum and the weaning were unaffected by treatment. Macroscopic examination, absolute and relative organ weights and histopathological evaluation of F0and F1parent animals showed no adverse treatment-related effects. It was concluded from these investigations that continuous treatment with sec-alkane sulfonate-sodium salts SAS at a level of 10000 ppm gave rise to a slight depression of somatic growth in parent animals and offspring in both generations, and to a marginal interference with survival of F1A offspring. At the intermediate dose level of continuous treatment (3000 ppm) slight depression of somatic growth of F1males was observed and there was marginal interference with survival of F1A and F2B offspring. At the lowest level of continuous treatment as well as in animals treated at all levels during organogenesis, no treatment-related adverse effects were observed. There were no indications for an embryotoxic or teratogenic effect related to treatment in any dose groups.