Endpoint study record: Toxicity to reproduction. 28-d combi-screening 500682

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Printing Date 2013-09-06 09:31:05 CEST

Restriction of specific regulatory purposes

Confidentiality

Owner (TS 204240) 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) / WIL Research Europe B.V. /

's-Hertogenbosch / Netherlands

Legal entity

owner

WIL Research Europe B.V. / 's-Hertogenbosch / Netherlands

Endpoint study record: Toxicity to reproduction. 28-d combiscreening 500682

UUID IUC5-ee1fc0f8-fee2-463c-aaa9-cb0fdce98acb

Dossier UUID 0

Author OTF / WIL Research Europe B.V. / 's-Hertogenbosch / Netherlands

Date 2013-09-05 14:21:12 CEST

Remarks

Administrative Data

Purpose flag key study; robust study summary

Study result type experimental result Study period 29 January - 18 March 2013

Reliability 1 (reliable without restriction)

Rationale for reliability incl.

deficiencies

This study has been performed according to OECD and/or EC guidelines and according to GLP

principles.

Data source

Reference

Reference

type

Author Year Title Bibliographic

source

Testing

laboratory

Report

no.

Owner

company

Company

study no.

Report

date

study

report

WIL

Research

Europe

B.V.

2013 Combined 28-day repeated

dose toxicity study with the

reproduction/developmental

toxicity screening test of

COPS_1 or CS-AHPS in

rats by oral gavage.

WIL Research

Europe B.V.,

Hambakenwetering

7, 5231 DD, ‘sHertogenbosch,

The Netherlands

500682 Society of

Chemical

Manufacturers

and Affiliates

1850 M Street,

NW Suite 700

WASHINGTON,

DC 20036, USA

2013-

07-15

Data protection claimed
Endpoint study record: Toxicity to reproduction. 28-d combi-screening 500682

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yes

Cross-reference to same study

See endpoint Study Record 7.5.1 and 7.8.2.

Materials and methods

Test type

other: combined repeated dose and reproduction / developmental screening

Limit test

no

Test guideline

Qualifier Guideline Deviations

according

to

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity

Screening Test)

no

according

to

other guideline: EPA, Health Effects Test Guidelines OPPTS 870.3650, Combined Repeated Dose Toxicity Study

with the Reproduction/Developmental Toxicity Screening Test, July 2000.

no

according

to

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test) no

according

to

other guideline: The United States EPA Health Effects Test Guidelines, OPPTS 870.3550,

Reproduction/Developmental Toxicity Screening Test, July 2000.

no

Principles of method if other than guideline

In addition, the procedures described in the report essentially conform to the following guidelines:

- Commission regulation (EC) No 440/2008 Part B: Methods for the Determination of Toxicity and other Health Effects; B.7: "Repeated

Dose (28 days) Toxicity (oral)". Official Journal of the European Union No. L142, May 2008.

- OECD Guidelines for Testing of Chemicals, Guideline 407, Repeated Dose 28-day Oral Toxicity Study in Rodents, October 2008.

- The United States EPA Health Effects Test Guidelines, OPPTS 870.3050, Repeated dose 28-day oral toxicity study in rodents, July

2000.

GLP compliance

yes

Test materials

Identity of test material same as for substance defined in section 1 (if not read-across)

yes

Test material identity

Identifier Identity

common name COPS_1 or CS-AHPS

Test material form

other: Liquid

Details on test material

- Identification: 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) 

- Name of test material (as cited in study report): COPS_1 or CS-AHPS

- Substance type: Clear colourless liquid

- Physical state: liquid

- Lot/batch No.: 12E1101

- Expiration date of the lot/batch: 13 August 2013

- Storage condition of test material: At room temperature in the dark

- Purity/composition correction factor required: No

- Hygroscopic: No

- Volatile: No

- Specific Gravity / Density: 1.17

- pH: 7 at concentration of 40.9%

- Stability in water: 1 year

- Solubility in water: 1-45%
Endpoint study record: Toxicity to reproduction. 28-d combi-screening 500682

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Confidential details on test material

None.

Test animals

Species

rat

Strain

other: Wistar (Han)

Sex

male/female

Details on test animals and environmental conditions

TEST ANIMALS

- Source: Charles River Deutschland, Sulzfeld, Germany.

Nulliparous and nonpregnant females and untreated animals were used at initiation of the study.

- Age at study initiation: Approximately 11 weeks.

- Weight at study initiation: mean weight at start of treatment was 322 gr (males) or 204 gr (females). 

- Fasting period before study: no

- Housing:

Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon cages.

Mating: Females were caged together with males on a one-to-one-basis in Macrolon cages.

Post-mating: Males were housed in their home cage with a maximum of 5 animals/cage. Females were individually housed in Macrolon

cages. Pups were kept with the dam until termination

General: Sterilised sawdust as bedding material and paper as cage enrichment were supplied. 

- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).

- Water: Free access to tap water.

- Acclimation period: At least 5 days 

Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40 to 70%, approximately 15 room air

changes/hour, and a 12-hour light/12-hour dark cycle. Any variations to these conditions were maintained in the raw data and had no

effect on the outcome of the study. 

IN-LIFE DATES

From: 29 January - 18 March 2013

Administration / exposure

Route of administration

oral: gavage

Vehicle

water

Details on exposure

- Method of formulation: Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenized to a visually

acceptable level. Adjustment was made for specific gravity of the test substance. No correction was made for the purity/composition of the

test substance.

- Storage conditions of formulations: At ambient temperature.

- Dose volume: 5 mL/kg body weight. Actual dose volumes were calculated according to the latest body weight.

Details on mating procedure

- M/F ratio per cage:1/1 (one female was cohabitated with one male of the same treatment group, avoiding sibling mating.

- Length of cohabitation: A maximum of 14 days was allowed for mating.

- Proof of pregnancy: Detection of mating was confirmed by evidence of sperm in the vaginal lavage and/or or by the appearance of an

intravaginal copulatory plug. This day was designated Day 0 post-coitum. Once mating had occurred, the males and females were

separated. 

- After successful mating each pregnant female was caged individually in Macrolon cages (MIII type, height 18 cm).

Analytical verification of doses or concentrations

yes

Details on analytical verification of doses or concentrations
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Analyses were conducted on a single occasion after the in-life phase (no validated analytical method was available during the in-life

phase) according to a validated method (Project 500674). Samples of formulations were analyzed for homogeneity (highest and lowest

concentration) and accuracy of preparation (all concentrations). The accuracy of preparation was considered acceptable if the mean

measured concentrations were 90-110% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was =

10%.

Duration of treatment / exposure

Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy. Females

were exposed for 42-48 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of

lactation (up to the day prior to scheduled necropsy). Two females of Group 1 and two females of Group 2 were not dosed during

littering..

Pups were not dosed directly but could have potentially be exposed to the test substance in utero, via maternal milk or from exposure to

maternal urine/faeces.

Frequency of treatment

Once daily for 7 d/w.

Details on study schedule

- Age at mating of the animals in the study: Approximately 13 weeks

Doses / concentrations

0, 100, 300, 1000 mg/kg bw/day

Basis actual ingested

No. of animals per sex per dose

10

Control animals

yes, concurrent vehicle

Further details on study design

- Dose selection rationale: Dose levels were based on information supplied by the sponsor.

- Selection of animals for selected measurements: 5 animals/sex/group were randomly selected at allocation for functional observations,

clinical pathology, macroscopic examination (full list), organ weights (full list) and histopathology. Selected females had live pups only for

Groups 1 and 2, and were randomly selected (as no litters with live pups) for Groups 3 and 4. 

- Parturition: The females were allowed to litter normally. Day 1 of lactation was defined as the day when a litter was found completed (i.e.

membranes, placentas cleaned up, nest built up and/or feeding of pups started). Females that were littering were left undisturbed.

- Identification of pups: On Day 1 of lactation, all pups were individually identified by means of subcutaneous injection of Indian ink.

Positive control

Not required.

Examinations

Parental animals: Observations and examinations

CAGE SIDE OBSERVATIONS

- Time schedule:At least twice daily. The time of death was recorded as precisely as possible.

DETAILED CLINICAL OBSERVATIONS

- Time schedule: Daily from treatment onwards up to the day prior to necropsy, detailed clinical observations were made in all animals.

Once prior to start of treatment and at weekly intervals during the treatment period this was also performed outside the home cage in a

standard arena. Clinical observations (clinical signs and arena) were conducted after dosing at no specific time point, but within a similar

time period after dosing for the respective animals. The time of onset, grade and duration of any observed sign was recorded. Signs were

graded for severity

BODY WEIGHT

- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females

were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum, and during lactation on Days 1 and 4.

FOOD CONSUMPTION

- Weekly, for males and females. Food consumption was not recorded during the mating period. Food consumption of mated females was

measured on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and on Days 1 and 4 of lactation.

FOOD EFFICIENCY: yes
Endpoint study record: Toxicity to reproduction. 28-d combi-screening 500682

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WATER CONSUMPTION

No. Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

OPHTHALMOSCOPIC EXAMINATION

No

HAEMATOLOGY

- Time schedule for collection of blood: immediately prior to scheduled post mortem examination.

- Anaesthetic used for blood collection: Yes (isoflurane)

- Animals fasted: Yes (with a maximum of 20 hours). Water was provided.

- How many animals: 5 animals/sex/group

- Parameters checked were: According to test guidelines

CLINICAL CHEMISTRY

- Time schedule for collection of blood: immediately prior to scheduled post mortem examination.

- Anaesthetic used for blood collection: Yes (isoflurane)

- Animals fasted: Yes (with a maximum of 20 hours). Water was provided.

- How many animals: 5 animals/sex/group

- Parameters checked were: According to test guidelines 

URINALYSIS

No

NEUROBEHAVIOURAL EXAMINATION

- Time schedule for examinations: The selected males were tested during Week 4 of treatment and the selected females were tested

during lactation (all before blood sampling).

- Dose groups that were examined: all

- Battery of functions tested: According to test guidelines 

GENERAL REPRODUCTION DATA

- Male number paired with, mating date, confirmation of pregnancy, and delivery day was recorded. 

- Pregnant females were examined to detect signs of difficult or prolonged parturition, and cage debris of these females was examined to

detect signs of abortion or premature birth. 

- Any deficiencies in maternal care (such as inadequate construction or cleaning of the nest, pups left scattered and cold, physical abuse

of pups or apparently inadequate lactation or feeding) were examined.

Estrous cyclicity (Parental animals)

Not determined.

Sperm parameters (Parental animals)

Slides of the testes were prepared for histopathological staging of spermatogenesis (5 males of the control and high dose group and

animals suspected to be infertile).

Litter observations

PARAMETERS EXAMINED

The following parameters were examined in F1 offspring:

number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross abnomalies, weight gain, physical or behavioural

abnormalities.

- Mortality: The numbers of live and dead pups on Day 1 of lactation and daily thereafter were determined. If possible, defects or cause of

death were evaluated.

- Clinical signs: At least once daily, detailed clinical observations were made in all animals.

- Body weights: Live pups were weighed on Days 1 and 4 of lactation.

- Sex: Sex was determined for all pups on Days 1 and 4 of lactation (by assessment of the ano-genital distance).

GROSS EXAMINATION OF DEAD PUPS

Yes, if possible, defects or cause of death were evaluated.

Postmortem examinations (Parental animals)

GROSS PATHOLOGY 

- All animals were fasted overnight (with a maximum of 24 hours) prior to planned necropsy, but water was provided. Animals surviving to

scheduled necropsy and all moribund animals were deeply anaesthetised and subsequently exsanguinated. 

- The number of former implantation sites and corpora lutea were recorded for all paired females.

- Selected 5 animals/sex/group and all animals that died spontaneously or were killed in extremis: According to test guidelines 

- All remaining animals and females which failed to deliver: According to test guidelines 
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ORGAN WEIGHTS

- Selected 5 animals/sex/group: According to test guidelines 

- All remaining males: Epididymides and Testes

HISTOPATHOLOGY

- According to test guidelines

Postmortem examinations (Offspring)

SACRIFICE

Pups surviving to planned termination were killed by decapitation on lactation Days 5-7.

GROSS NECROPSY

All pups were sexed and descriptions of all external abnormalities were recorded. The stomach was examined for the presence of milk. If

possible, defects or cause of death were evaluated. Any abnormal pup, organ or tissue was preserved in 10% buffered formalin for

possible further examination.

HISTOPATHOLOGY / ORGAN WEIGHTS

No.

Statistics

The following statistical methods were used to analyze the data:

- If the variables could be assumed to follow a normal distribution, the Dunnett-test (many-to-one t-test) based on a pooled variance

estimate was applied for the comparison of the treated groups and the control groups for each sex.

- The Steel-test (many-to-one rank test) was applied if the data could not be assumed to follow a normal distribution.

- The Fisher Exact-test was applied to frequency data.

- The Kruskal-Wallis nonparametric ANOVA test was applied to motor activity data to determine intergroup differences. In case intergroup

differences were seen, the Wilcoxon test was applied to compare the treated groups to the control group.

All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance. Group means were calculated for

continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the

basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off

before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics

values.

Reproductive indices

For each group, the following calculations were performed:

- Mating index: Number of females mated/Number of females paired x 100

- Fertility index: Number of pregnant females/Number of females paired x 100

- Conception index: Number of pregnant females/Number of females mated x 100

- Gestation index: Number of females bearing live pups/Number of pregnant females x 100

- Duration of gestation: Number of days between confirmation of mating and the beginning of parturition

Offspring viability indices

- Percentage live males at First Litter Check: Number of live male pups at First Litter Check/Number of live pups at First Litter Check x

100

- Percentage live females at First Litter Check: Number of live female pups at First Litter Check/Number of live pups at First Litter Check

x 100

- Percentage of postnatal loss Days 0-4 of lactation: Number of dead pups on Day 4 of lactation/Number of live pups at First Litter Check

x 100

- Viability index: Number of live pups on Day 4 of lactation / Number of pups born alive x 100

Results and discussions

Effect levels

Endpoint Generation Sex Effect

level

Based

on

Basis for effect level / Remarks

NOAEL P male/female >= 1000

mg/kg

bw/day

(actual

dose

received)

test

mat.

No parental toxicity was observed up to the highest dose level tested. 

NOAEL F1 male/female < 100

mg/kg

bw/day

(actual

test

mat.

A reproduction NOAEL could not be determined, as a possible treatmentrelated effect on reproduction at this dose could not be excluded (One female

was found not pregnant, and one female had a total litter loss (her single pup

died on Day 5 of lactation. Two females showed a reduced number of corpora
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dose

received)

lutea and implantation sites).

Results of examinations: parental animals

Clinical signs (parental animals)

no effects

Body weight and food consumption (parental animals)

no effects

Test substance intake (parental animals)

not examined

Reproductive function: estrous cycle (parental animals)

not examined

Reproductive function: sperm measures (parental animals)

not examined

Reproductive performance (parental animals)

yes

Organ weights (parental animals)

no effects

Gross pathology (parental animals)

no effects

Histopathology (parental animals)

no effects

Details on results (parental animals)

MORTALITY

No mortality occurred during the study period. One female at 100 mg/kg was sacrificed before scheduled necropsy due to total litter loss

on Day 5 of lactation.

CLINICAL SIGNS

No clinical signs of toxicity were noted during the observation period. For one female at 300 mg/kg blood coming out of the vagina was

recorded on the scheduled day of necropsy. This was considered to be related to the observation of a large fetus at necropsy of this

same animal, and no such finding was noted for any of the other females. The incidence of alopecia and scabs occurred within the range

of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study. At

the incidence observed, these were considered signs of no toxicological relevance.

FUNCTIONAL OBSERVATIONS

Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all animals tested. No toxicologically relevant

changes in motor activity were noted. For motor activity, all groups showed a similar habituation profile with high activity in the first interval

that decreased over the duration of the test period. The higher motor activity of females at 300 and 1000 mg/kg (statistically significant at

1000 mg/kg) was considered due to these females not having delivered offspring. This variation was therefore considered to be of no

toxicological relevance.

BODY WEIGHTS

No toxicologically relevant changes in body weights and body weight gain were noted. The lower body weight gain of the two females at

300 mg/kg found pregnant (based on implantation sites) and two females at 100 mg/kg during the end of the post-coitum period was

considered to be due to a small litter size or absence of litters, and was therefore not considered toxicologically relevant. The pregnant

females at 100 mg/kg showed a normal body weight gain during the complete study period. No data on body weight gain during lactation

was obtained at 300 and 1000 mg/kg as no litters were born in these groups.

FOOD CONSUMPTION

No toxicologically relevant changes in food consumption before or after correction for body weight was noted. The lower food intake of the

two females at 300 mg/kg found pregnant based on implantation sites (but no offspring) and two females at 100 mg/kg during the end of

the post-coitum period (and for one of these two females also during lactation) was considered to be due to a small litter size or absence

of litters, and was therefore not considered toxicologically relevant.

HAEMATOLOGY
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No toxicologically relevant changes were noted in haematological parameters. The lower reticulocyte counts at 300 and 1000 mg/kg were

considered to be due to the pregnancy status (small litter size or absence of litters), and to be within the physiological range of nonpregnant females. The higher relative neutrophil counts with concurrently lower relative lymphocyte counts for one female at 1000 mg/kg

was considered to be a secondary non-specific response to stress, and similar changes in other animals of the same group were absent.

The statistically significantly higher activated partial thromboplastin time (APTT) for females at 300 mg/kg occurred in the absence of a

dose-related trend. The higher reticulocyte, red cell distribution width (RDW) and/or mean corpuscular volume (MCV) values shown by two

females at 100 mg/kg were not seen for other animals of this dose group. These changes were therefore considered to be of no

toxicological relevance.

CLINICAL BIOCHEMISTRY

No toxicologically relevant changes were noted in clinical biochemistry parameters. All statistically significant changes in clinical

biochemistry parameters in males at 100 and 300 mg/kg were considered to be of no toxicological significance as they occurred in the

absence of a doserelated trend and remained within the range considered normal for rats of this age and strain. These changes consisted

of higher total bilirubin, creatinine, bile acid and sodium levels and lower cholesterol levels. The statistically significantly changes for

females at 300 and 1000 mg/kg were considered to have occurred due to the pregnancy status of these females (small litter size or

absence of litters), and means remained within the range of values expected for non-pregnant females of a similar age and same strain.

These changes consisted of a lower alanine aminotransferase activity level (ALAT), higher albumin, sodium and chloride level, and lower

urea level. The lower sodium level of females at 100 mg/kg remained within the range considered normal for rats of this age and strain.

These changes were therefore considered to be of no toxicological relevance.

MACROSCOPIC EXAMINATION

Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a direct result of treatment.

The large fetus noted in utero of one female at 300 mg/kg was considered due to it being the only fetus growing larger than normal

compared to fetuses from a larger litter. This was therefore considered to be an indirect effect of treatment. The incidence of other

necropsy findings among control and treated animals was within the background range of findings that are encountered among rats of this

age and strain, and did not show a dose-related incidence trend. No toxicological relevance was therefore ascribed to these findings.

ORGAN WEIGHTS

No toxicologically relevant changes were noted in organ weights and organ to body weight ratios. The statistically significant lower liver

weight, and higher thymus and ovary weight of females at 300 and 1000 mg/kg were considered to be due to their pregnancy status

(small litter size or absence of litters). The statistically significantly higher thyroid to body weight ratio of females at 1000 mg/kg remained

within the range considered normal for rats of this age and strain, and occurred in the absence of correlating treatment-related

histopathology findings. These changes were therefore not considered toxicologically relevant.

MICROSCOPIC EXAMINATION

No toxicologically relevant microscopic findings were noted. The microscopic findings for one female treated at 1000 mg/kg were regarded

to be related to a gavage accident, because of the single incidence and the recorded disruption of the normal glandular stomach

architecture. The remainder of microscopic findings recorded were within the normal range of background pathology encountered in

Wistar (Han) rats of this age. No histological correlation could be made for the lack of offspring. All females showed a normal

reproductive cycle. For the males, no abnormalities were seen in the reproductive organs which could account for their infertility.

Furthermore, the spermatogenic staging profiles were normal for all examined males. 

REPRODUCTION DATA

Reproduction parameters were affected at all dose levels. At 1000 mg/kg, none of the females were found pregnant. At 300 mg/kg, only

two females were found pregnant based on presence of implantation sites, resulting in a reduced fertility and conception index. The two

pregnant females showed a reduced number of corpora lutea and/or implantation sites. At 100 mg/kg, nine females were found pregnant.

One of these females was found pregnant based on presence of implantation sites. One female was found not pregnant, and one female

had a total litter loss (her single pup died on Day 5 of lactation). Two of those females showed a reduced number of corpora lutea and

implantation sites. Other pregnant females of this dose group showed a normal number of corpora lutea and implantation sites. Mating

index and precoital time were unaffected by treatment up to 1000 mg/kg.

Results of examinations: offspring

Viability (offspring)

no effects

Clinical signs (offspring)

no effects

Body weight (offspring)

no effects

Sexual maturation (offspring)

not examined

Organ weights (offspring)

not examined
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Gross pathology (offspring)

no effects

Histopathology (offspring)

not examined

Details on results (offspring)

DEVELOPMENTAL DATA

No toxicologically relevant effects on gestation index and duration, parturition, maternal care and early postnatal pup development

(mortality, clinical signs, body weight and macroscopy) were observed at 100 mg/kg. At 300 and 1000 mg/kg no pups were delivered, and

hence developmental effects could not be assessed for these dose levels.

MORTALITY PUPS

One pup of the control group was missing (most likely cannibalised) on Day 2 of lactation, three pups at 100 mg/kg were found dead at

first litter check, and one pup at 100 mg/kg was found dead on Day 5 of lactation. No toxicological relevance was attributed to these

dead/missing pups since the mortality incidences remained within the range considered normal for pups of this age.

OBSERVATIONS PUPS

The one pup of a litter at 100 mg/kg that was found dead on Day 5 of lactation showed a lean appearance from first litter check onwards

and absence of milk in the stomach from Day 2 onwards. In addition, this pup showed a slight weight loss over Days 1-4 of lactation. At

this single occurrence, it was not considered toxicologically significant.

Incidental findings consisted of a wound, scabbing, absence of milk in the stomach, blue spot, and abnormal posture of the hindleg. The

pups that were found dead at first litter check showed severe autolysis. The nature and incidence of these findings remained within the

range considered normal for pups of this age, and were therefore considered to be of no toxicological relevance. Absence of milk in the

stomach was noted for one litter of the control group. This finding for these control pups surviving to the scheduled necropsy is likely

secondary to the time when the actual necropsies were performed, and not considered toxicologically relevant.

Any other information on results incl. tables

The concentrations analysed in the formulations of Group 2, Group 3 and Group 4 were in agreement with target concentrations (i.e.

mean accuracies between 90% and 110%). No test substance was detected in the Group 1 formulations. The formulations of Group 2

and Group 4 were homogeneous (i.e. coefficient of variation = 10%).

Overall remarks, attachments

Remarks on results including tables and figures

Parental results:

No toxicologically relevant findings were observed up to the highest dose level tested (1000 mg/kg). Several effects considered to be

secondary to a small litter size or absence of litters were noted for females at 300 and 1000 mg/kg in motor activity, body weight gain,

food consumption, clinical biochemistry parameters, organ weights and microscopic examinations. No direct treatment-related changes

were noted in any of the parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weight, food

consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).

Reproductive results:

At 1000 mg/kg, none of the females were found pregnant, and at 300 mg/kg only two females were found pregnant (based on presence

of implantation sites) and showed a reduced number of corpora lutea and implantation sites. At 100 mg/kg, nine females were found

pregnant (one of these based on presence of implantation sites). One female was found not pregnant, and one female had a total litter

loss (her single pup died on Day 5 of lactation). Two females showed a reduced number of corpora lutea and implantation sites, while

other pregnant females of this dose group showed a normal number of corpora lutea and implantation sites. As these findings at 100

mg/kg were in line with the affected reproduction noted at 300 and 1000 mg/kg, a possible treatment related effect could not be excluded.

Mating index and precoital time were normal up to the highest dose level tested (1000 mg/kg).

Developmental results:

No developmental toxicity was observed at 100 mg/kg in any of the developmental parameters investigated in this study (i.e. gestation

index and duration, parturition, maternal care and early postnatal pup development consisting of mortality, clinical signs, body weight and

macroscopy). Developmental effects could not be assessed at 300 and 1000 mg/kg as no pups were born.

Applicant's summary and conclusion

Conclusions

In this OECD 422 study, treatment with COPS_1 or CS-AHPS by oral gavage in male and female Wistar Han rats at dose levels of 100,
Endpoint study record: Toxicity to reproduction. 28-d combi-screening 500682

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300 and 1000 mg/kg revealed no parental toxicity up to 1000 mg/kg. Reproduction toxicity was observed at all dose levels. No

developmental toxicity was observed at 10 mg/kg. However, developmental toxicity could not be assessed at 300 and 1000 mg/kg since

no pups were born.

Based on these results, the following No Observed Adverse Effect Levels (NOAEL) were derived: 

Parental NOAEL: at least 1000 mg/kg (409 mg active ingredient/kg); 

Reproduction NOAEL: could not be determined (below 100 mg/kg (41 mg active ingredient/kg)); 

Developmental NOAEL: at least 100 mg/kg (41 mg active ingredient/kg)