Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

According to the criteria of Regulation (EC) No 1907/2006, Annex IX, 8.7.3, column 1, an extended one-generation reproductive toxicity study is not indicated as the results of the 90-day repeated dose toxicity study and the 2-years repeated dose toxicity study with the registered substance do not demonstrate any adverse effects on reproductive organs or tissues. In addition, the substance has only a low potential for absorption as reviewed in the toxicokinetic assessment. The effects observed from acute and repeated dose toxicity studies are of local characteristic and reflect the irritant nature of the test substance which was demonstrated by skin and eye irritation studies.

Effects on developmental toxicity

Description of key information
Developmental toxicity NO(A)EL >= 250 mg/kg bw/day (OECD 414). 
Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04 Aug - 05 Dec 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted 22 January 2001
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted August 1998
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries Testing guidelines for Toxicology studies, 12 NohSa No 8147, 24 November 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl:CD® (SD)IGS BR
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Margate, UK
- Age at study initiation: not specified in report, animals purchased time-mated
- Weight at study initiation: 197-268 g (females on arrival, prior to GD3)
- Housing: individual in solid-floor propylene cages with stainless steel lids in a single air-conditioned room, bedding with softfood flakes
- Diet: pellet diet (Rodent 2018C Teklad Global Certified Diet, Harlan; Oxon; UK) ad libitum, environmental enrichment provided in the form of wooden chew blocks and cardboard fun tunnels
- Water: ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 50 ± 20
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 04 Aug 2013 (first day of treatment) To: 22 Aug 2013 (final day of necropsy)
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distilled water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
the test item was prepared at the appropriate concentrations as a solution in distilled water.

VEHICLE
- Concentration in vehicle: 6, 20 and 50 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test item concentration in the test samples was determined by high performance liquid chromatography (HPLC/UV) using an external standard technique.The formulations investigated during the study were found to comprise test item in the range of 101% to 104% and, thus, the required content limit ± 10% with reference to the nominal content was met. The test item was found to be stable in the formulations when kept for twenty one days in the refrigerator due to results which met the variation limit of 10% from the time-zero mean. Thus, the results indicate the accurate use of the test item and distilled water as vehicle during this study. The formulations were found to be homogeneously prepared, and sufficient formulation stability under storage conditions was approved.
Details on mating procedure:
- Impregnation procedure: purchased timed pregnant
Duration of treatment / exposure:
Gestation Day 5-19
Frequency of treatment:
daily, 7 days/week
Duration of test:
until Gestation Day 20 (terminal sacrifice)
Remarks:
Doses / Concentrations:
30, 100 and 250 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
24 P females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were chosen based on previous toxicity data.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily
- Cage side observations checked included: following arrival, all animals were examined for overt signs of toxicity, ill-health or behavioural changes once daily during the gestation period. An additional observation was also performed five hours after dosing during the normal working week. All observations were recorded.

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: individual body weights were recorded for each surviving individual animal at Day 3, 5, 6, 7, 8, 11, 14, 17 and 20 (termination kill) of gestation.

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined: Yes, at Day 3, 5, 8, 11, 14, 17 and 20 of gestation.

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: daily by visual inspection of water bottles
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: ovaries and uteri of pregnant females, full external and internal examination; any macroscopic abnormalities were recorded; the stomach was retained from all females
Ovaries and uterine content:
The ovaries and uterine content were examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

- Other: fetal sex, placental weight, position and type of intrauterine implantation
Fetal examinations:
- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [all per litter]
- Skeletal examinations: Yes: [all per litter]
- Head examinations: No
Statistics:
Group mean values were calculated to include data from all females with live fetuses on Day 20 of gestation. As the litter was the standard unit of assessment, values were first calculated within the litter, and group mean values represent the mean of these individual litter values.
The following parameters were analyzed statistically, where appropriate, using the test methods outlined below:
Female body weight change, food consumption and gravid uterus weight: Bartlett´s test for homogeneity of variance and one way analysis of variance, followed by Dunnett´s multiple comparison test or, if unequal variances were observed, an alternative multiple comparison test.
All caesarean necropsy parameters and fetal parameters: Kruskal-Wallis non-parametric analysis of variance; and a subsequent pairwise analysis of control values against treated values using the Mann-Whitney ‘U’ test, where significance was seen.
Fetal evaluation parameters, including skeletal or visceral findings: Kruskal-Wallis non-parametric analysis of variance and Mann-Whitney ‘U’ test.
Probability values (p) demonstrating different levels of significance were chosen as follows:
p<0.001 ***
p<0.01 **
p<0.05 *
p>=0.05 (not significant)
Indices:
Pre implantation loss: [(number of corpora lutea-number of implantations) / number of corpora lutea] x 100;
Post-implantation loss: [(number of implantations–number of live foetuses)/number of implantations] x 100;
Sex ratio: % male fetuses (sex ratio)= [Number of male foetuses / Total number of foetuses] x 100

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
250 mg/kg bw/day: the female found dead on Day 10 did not show any clinical signs prior to Day 10. The female found dead on Day 18 had noisy respiration on Days 10 and 17 and increased salivation and pilo-erection on Day 17.
5/22 females showed incidences of increased salivation between Days 10 and 19 and 3/22 females also showed noisy respiration on either Day 6 or 7.

100 mg/kg bw/day: 1/24 females showed increased salivation on Day 18 and 1/24 females had noisy respiration on Day 13.

Observations of this nature are commonly observed following the oral administration of an unpalatable or irritant test item formulation and are considered not to represent true systemic toxicity.

Control: 1/24 females had fur loss on Day 18. In the absence of treatment this was considered of no toxicological importance.
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, treatment-related
Description (incidence):
250 mg/kg bw/d: One female treated was found dead on Day 10 and another female was found dead on Day 18.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
250 mg/kg bw/day: body weight gain of the females was lower throughout the treatment period and cumulative body weight gain to Day 14 (where body weight is not greatly influenced by the weight of the gravid uterus) was 24% lower than the corresponding control value. By termination on Day 20, cumulative body weight gain was 15% lower than the corresponding control values. When final body weight was adjusted for the gravid uterus, an overall body weight gain reduction was evident during gestation.

100 mg/kg bw/day: females showed a slight reduction in cumulative body weight gain from Day 7 onwards. When final body weight was adjusted for the gravid uterus, an overall body weight gain reduction was evident during gestation.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
250 mg/kg bw/day: females showed a reduction in food consumption throughout the treatment period. Statistically significant reductions were evident between Days 8 and 11 (p < 0.001) and Days 14-17 (p< 0.05) compared with the control group.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Description (incidence and severity):
The female treated with 250 mg/kg bw/day that was found dead on Day 10 had gaseous distention in the stomach and gastro-intestinal tract. The other interim death female had sloughing on the non-glandular region of the stomach, seven dead foetuses in the right uterine horn and five dead foetuses in the left uterine horn. The remaining females treated with 250 mg/kg bw/day that were terminated on Day 20 all had sloughing on the non-glandular region of the stomach.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
not examined
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Dose descriptor:
LOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
effects observed, treatment-related
Description (incidence and severity):
Females from all treatment groups showed a statistically significant reduction (p<0.05 - p<0.01) in the number of corpora lutea. The intergroup differences did not show a true dose related response and were considered to be incidental and unrelated to treatment due to ovulation and mating occurring prior to the administration of the test item. As a consequence of incidentally lower number of corpora lutea in treated females litter size was statistically significantly reduced (p<0.05) in all treated females. The intergroup differences did not show a true dose related response and in the absence of any effect on post-implantation loss and mean fetal weights it supports the assumption that the intergroup differences in litter size was considered to be incidental and of no toxicological importance.
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
There was no obvious adverse effect of maternal treatment on litter data as assessed by the mean number of implantations, early and late embryonic/fetal deaths and live fetuses or sex ratio, as assessed by percentage male.
For all dose groups, there were no significant treatment-related trends in the proportion of fetuses (or litters) with evidence of visceral or skeletal anomalies. The types of external, visceral and skeletal anomalies were those commonly observed for this type of study. There were no findings that were considered to represent any known malformations.
Dose descriptor:
NOEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: No adverse effect on developmental toxicity observed at the highest tested dose level.
Abnormalities:
not specified
Developmental effects observed:
not specified

All treated females showed statistically significant reduction of corpora lutea and in consequence reduced litter sizes. The intergroup differences did not show a true dose related response and were considered to be incidental and unrelated due to ovulation and mating occurring prior to the administration of the test item starting on GD5. The absence of any effect on post-implantation loss and mean fetal weights supports the assumption that these intergroup differences in litter size were considered to be incidental and of no toxicological importance. Foetuses of the 250 mg/kg bw/day group showed a statistically significant increase in the percent of foetuses showing one or more ossified forepaw phalanges. Due to the isolated occurrence of this observation it is not considered to be a true developmental effect and it is therefore of no biological importance.

Table A: Group Mean Litter Data Values (Mean ± SD)

Dose Level (mg/kg bw/day)

Number of Litters

Number of Corpora Lutea

Number of Implants

Number of Embryonic/Fetal Deaths

Number of Live Implants

Early

Late

Total

Male

Female

Total

0 (control)

23

15.2 ± 1.8

13.9 ± 1.6

0.1 ± 0.3

0.1 ± 0.6

0.3 ± 0.7

6.9 ± 1.8

6.8 ± 1.9

13.7 ± 2.1

30

21

14.3* ± 1.8

13.1 ± 2.0

1.0 ± 2.4

0.1 ± 0.4

1.0 ± 2.5

6.0 ± 2.7

6.1 ± 2.4

12.1* ± 3.4

100

24

13.7* ± 1.9

13.0 ± 1.8

0.6 ± 1.3

0.2 ± 0.7

0.8 ± 1.4

6.5 ± 2.1

5.6 ± 1.7

12.1* ± 2.5

250

22

13.9** ± 1.2

13.2 ± 1.1

0.5 ± 1.4

0.2 ± 0.9

0.7 ± 1.6

6.5 ± 2.0

6.0 ± 1.9

12.5* ± 2.2

p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 not significant

Table A: Group Mean Litter Data Values (continued) (Mean ± SD)

Dose Level (mg/kg bw/day)

Implantation Loss (%)

Male foetuses (%)

Mean Male Fetal Weight (g)

Mean Female Fetal Weight (g)

Mean Fetal Weight (g)

Mean Placental Weight (g)

Litter Weight (g)

Total Placental Weight (g)

Pre

Post

0 (control)

8.4 ± 7.4

2.3 ± 7.2

50.4 ± 11.3

4.117 ± 0.260

3.935 ± 0.264

4.026 ± 0.257

0.564 ± 0.047

54.883 ± 8.672

7.667 ± 1.141

30

8.0 ± 10.0

8.4 ± 19.4

48.6 ± 15.2

4.152 ± 0.405

3.881 ± 0.372

4.005 ± 0.365

0.578 ± 0.155

48.345 ± 13.761

6.729 ± 1.942

100

5.0 ± 5.2

6.9 ± 12.2

53.7 ± 12.8

4.166 ± 0.339

3.886 ± 0.269

4.036 ± 0.289

0.542 ± 0.060

49.142 ± 11.318

6.540 ± 1.503

250

4.8 ± 5.3

5.5 ± 12.5

52.2 ± 13.7

4.221 ± 0.347

4.033 ± 0.327

4.126 ± 0.320

0.540 ± 0.057

51.237 ± 8.200

6.744 ± 1.369

 

Table B: Group Mean Cumulative Body Weight Change Values (Mean ± SD)

Dose Level (mg/kg bw/day)

Number of Animals

Cumulative Body Weight Change (g) from GD5

GD6

GD7

GD8

GD11

GD14

GD17

GD20

0 (control)

23

4.0 ± 3.4

7.6 ± 3.7

13.0 ± 5.1

33.6 ± 6.6

51.6 ± 8.1

82.3 ± 10.4

127.8 ± 14.9

30

21

5.1 ± 5.4

7.1 ± 8.2

12.9 ± 10.2

34.0 ± 12.9

53.5 ± 15.5

82.2 ± 19.2

127.0 ± 23.4

100

24

2.3 ± 5.3

6.1 ± 5.5

8.8 ± 7.5

27.6 ± 9.9

44.8 ± 13.8

74.2* ± 12.4

114.3* ± 17.2

250

24/22+

0.5 ± 9.0

3.6 ± 10.1

7.3 ± 11.2

21.7** ± 14.6

39.3* ± 17.8

68.2** ± 20.0

109.1** ± 24.7

+ = n=23 on GD10, n=22 on GD18
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 not significant

 

Table C: Group Mean Gravid Uterus Weight , Adjusted Body Weight and Body Weight Change Values (Mean ± SD)

Dose Level (mg/kg bw/day)

Number of Animals

Body Weight (g) on Days of Gestation

Body Weight Change (g) during GD5-20

Gravid Uterus Weight (g)

Adjusted Body Weight (g) GD20

Adjusted Body Weight (g) Change GD5-20

GD5

GD20

0 (control)

23

262.6 ± 20.6

390.4 ± 29.8

127.8 ± 14.9

84.663 ± 12.502

305.8 ± 23.4

43.2 ± 8.2

30

21

261.7 ± 12.9

288.7 ± 27.3

127.0 ± 23.4

75.544 ± 18.733

313.2 ± 28.6

51.5 ± 22.7

100

24

260.6 ± 14.4

374.9 ± 24.8

114.3** ± 17.2

75.893 ± 15.615

299.0 ± 20.7

38.4* ± 14.6

250

24/22+

257.6 ± 15.5

368.1 ± 32.4

109.1** ± 24.7

77.535 ± 11.935

290.6 ± 25.0

31.6** ± 18.7

+ = n=23 on GD10, n=22 on GD18
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 not significant

 

Table D: Group Mean Food Consumption Values (Mean ± SD)

Dose Level (mg/kg bw/day)

Number of Animals

Food Consumption (g/rat/day) between Days of Gestation

3-5

5-8

8-11

11-14

14-17

17-20

0 (control)

23

23.0 ± 3.1

25.1 ± 2.5

26.5 ± 2.6

26.1 ± 2.8

25.9 ± 3.0

28.0 ± 2.4

30

21

23.6 ± 3.7

24.5 ± 3.9

27.1 ± 3.7

26.5 ± 3.7

26.0 ± 4.2

28.7 ± 3.8

100

24

22.8 ± 2.9

23.9 ± 2.8

24.9 ± 3.3

24.6 ± 3.7

24.5 ± 3.4

27.5 ± 2.9

250

24/22+#

22.3 ± 3.4

22.4 ± 5.3

21.5*** ± 4.7

23.3 ± 4.3

22.6* ± 4.2

25.4 ± 3.8

+ = n=23 on GD10, n=22 on GD18
# = Food consumption not performed on GD 8 and 11 for female Nos. 85 to 90 due to technician error
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 not significant

In conclusion the oral administration of the test substance to pregnant rats by oral gavage during gestation at dose levels of 30, 100 and 250 mg/kg bw/day resulted in treatment related effects detected in females treated with 250 and 100 mg/kg bw/day. The NOAEL was therefore considered to be 30 mg/kg bw/day.

No toxicological significant changes were detected in the offspring parameters measured. The NOEL for developmental toxicity was therefore considered to be 250 mg/kg bw/day. 

Conclusions:
Under the conditions of the study the test substance did not induce any treatment-related biologically relevant malformations in the developing unborn organism in the presence of maternal toxicity. Therefore, the test substance does not meet the criteria for classification for prenatal developmental toxicity according to Regulation (EC) No 1272/2008. The available data is thus conclusive but not sufficient for classification.
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 Nov 2016 - 15 March 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
adopted Jan 2001
Deviations:
yes
Remarks:
(purity of the test material not given)
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Deviations:
not specified
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries, Test Data for Registration of Agricultural Chemicals, 12 Nohsan No. 8147
Version / remarks:
24 Nov 2000
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Remarks:
The Department of Health of the Government of the United Kingdom
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS
- Age at study initiation: 18 - 22 weeks
- Weight at study initiation: 2.31 - 4.36 kg
- Housing: The animals were individually housed during acclimatization and gestation period, and co-housed (M/F ration: 1/1) during mating period in suspended cages fitted with perforated floor panels. An aspen chew block was provided to each cage as environmental enrichment.
- Diet: Teklad 2930 pelleted diet, initially 150 g/animal/day during acclimatization up to one week prior to the onset of mating and 200 g/animal/day thereafter. In addition, a small supplement of autoclaved hay was given on a daily basis and a small amount of chopped fresh vegetables were given twice weekly.
Water: Potable water, ad libitum
- Acclimation period: 19 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 15 - 21
- Humidity (%): 45 - 70
- Photoperiod (hrs dark / hrs light): 10/14

IN-LIFE DATES: From: 23 Nov 2016 To:
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
purified
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Dosing solutions were prepared weekly by dissolving appropriate amounts of the test material in purified water yielding a final concentration of 100 mg/mL. The remaining concentrations were prepared by serial dilution with further quantities of vehicle.

VEHICLE
- Amount of vehicle (if gavage): 5 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability of the test substance in water formulations at nominal concentrations of 2 mg/mL and 200 mg/mL was confirmed following refrigeration for 17 days at the same testing facility (Study No. KK40KJ)
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1
- Verification of same strain and source of both sexes: yes
- Other: The day of mating is considered to be Day 0 of Gestation. Where possible only females mating at least twice were allocated.
Duration of treatment / exposure:
Day 6 - 28 of gestation
Frequency of treatment:
daily, 7 days/week
Duration of test:
Day 29 post mating
Dose / conc.:
125 mg/kg bw/day (nominal)
Dose / conc.:
250 mg/kg bw/day (nominal)
Dose / conc.:
500 mg/kg bw/day (nominal)
No. of animals per sex per dose:
22
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Doses were chosen based on the results of a preliminary study for effects on embryo-fetal development in the New Zealand White rabbit at the same testing facility (Study No. SF87BJ). In that study, treatment with the test substance at 125 or 250 mg/kg/day was well-tolerated, with no effects apparent that precluded the use of these doses on the subsequent main study for effects on embryo-fetal development. Treatment at 500 mg/kg/day elicited effects of reduced body weight gain and food consumption on the does, and reduced litter and fetal weights amongst the litters of females which received the test substance at 500 mg/kg/day, the magnitude of which did not preclude the use of this dose on the subsequent main study for effects on embryo-fetal development where the highest dose was chosen with the aim to induce some developmental and/or maternal toxicity. Doses of 125, 250 and 500 mg/kg/day were considered appropriate for the subsequent main study for effects on embryo-fetal development.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes, all animals were inspected visually for evidence of clinical signs and mortality.
- Time schedule: At least twice daily

DETAILED CLINICAL OBSERVATIONS: Yes, a detailed phyisical examination was performed on all animals to monitor general health.
- Time schedule: On Days 0, 6, 12, 18, 24 and 29 after mating

BODY WEIGHT: Yes, body weights of all animals were recorded.
- Time schedule for examinations: Weekly during acclimatizatuin, on the day of mating and on Days 0, 3 and 6 - 29 post mating.

FOOD CONSUMPTION: Yes
- The weight of food supplied to each animal, that remaining and an estimate of any spilled was recorded daily from Day 1 after mating.

POST-MORTEM EXAMINATIONS: Yes, all adult animals were subject to a detailed necropsy. Animals surviving until the end of the scheduled study period were killed on Day 29 post mating. Females that exhibited pregnancy loss were killed on the day of detection.
- Organs examined: All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative.
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes, for females surviving to term.
- Number of corpora lutea: Yes, for all animals.
- Number of implantations: Yes, for all animals.
- Number of early resorptions: Yes, for all animals.
- Number of late resorptions: Yes, for all animals.
Fetal examinations:
- External examinations: Yes, all fetuses were weighed, and were subject to an external examination and a gross internal examination of the viscera of the neck, thorax and abdominal cavities and the sex of each fetus was recorded.
- Soft tissue examinations: Yes, serial sections were examined for soft tissue abnormalities.
- Skeletal examinations: Yes, the torsos of the decapitated fetuses and the remaining fetuses were eviscerated and fixed in Industrial Methylated Spirit.
- Head examinations: Yes, half per litter were decapitated and the heads were fixed in Bouin`s fluid.
Statistics:
The following data types were analysed at each timepoint separately:
- Body weight, using absolute values and gains over appropriate study periods
- Gravid uterine weight and adjusted body weight
- Food consumption, over appropriate study periods
- Litter size and survival indices
- Fetal, placental and litter weight

Detailed information on statistical information is provided under "any other information on materials and methods incl. tables".
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
125 mg/kg bw/d: 1/22 females had shown swaying and gasping respiration prior to death, and was found dead on Day 15 of gestation.
500 mg/kg bw/d: 1/22 females showed signs of respiratory distress and was getting very stressed when the technicians tried to touch her. The animal was despatched to necropsy on Day 9 of gestation (plase refer to "Mortality").
Dermal irritation (if dermal study):
not examined
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Control: 1/22 females was found dead on Day 21 of gestation. The animal did not show any clinical signs prior to death, and there were no abnormalities detected at macroscopic examination.

125 mg/kg bw/d: 1/22 females was found dead on Day 15 of gestation. Clinical signs and a loss of body weight were observed prior to death. Macroscopic examination revealed abnormalities (please refer to "Gross pathological findings"). However, according to the study director the death was unclear and was not considered to be treatment-related. 1/22 females was despatched to necropsy on Day 24 of gestation following evidence of abortion. No clinical signs were observed prior to death and no abnormalities were detected at macroscopic examination.

500 mg/kg bw/d: 1/22 females was despatched to necropsy on Day 9 of gestation having shown signs of respiratory distress and a lost 410 g of body weight since the start of dosing. Macroscopic examination revealed abnormalitites (please refer to "Gross pathological findings"). According to the study director, the death was considered to be non-treatment related.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
There was no clear effect of treatment on the body weight performance of females receiving the test substance at 125 or 250 mg/kg bw/d when compared with that of the control group. The mean body weight loss of females receiving dosed at 500 mg/kg bw/d was slightly greater than that of the control group after the start of treatment (between Days 6 and 8 after mating), and body weight gain was slightly less than that of the control animals from Days 8 - 29 after mating.
When mean values of body weight and body weight gain were adjusted for the contributions of the gravid uterus, overall maternal mean body weight loss during Days 6 - 29 of gestation was greater at 500 mg/kg bw/d than in the control group.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
There was no clear effect of treatment on the food consumption of females receiving the test substance at 125 or 250 mg/kg bw/d when compared with that of the control group. The food consumption of females dosed at 500 mg/kg bw/d was slightly lower than that of the control group from the start of treatment (Day 6 after mating), resulting in a lower overall (Day 6 - 28 after mating) food consumption.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
125 mg/kg bw/d: The macroscopic examination of the animal found dead on Day 15 of gestation, revealed discoloured (dark) adipose tissue in the thoracic region, dark staining to the muzzle, dark fluid in the nasal turbinates, and dark right lungs.
500 mg/kg bw/d: The animal which was despatched to necropsy on Day 9 of gestation had poorly defined, dark areas on the lungs, gas in the jejunum and duodenum, and findings in the stomach consisting of a raised and firm antrum with a depressed area on the glandular mucosa at necropsy.

These findings were not considered to be treatment related or toxicologically relevant.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Number of abortions:
effects observed, non-treatment-related
Description (incidence and severity):
125 mg/kg bw/d: The animal which was despatched to necropsy on Day 24 of gestation following evidence of abortion, was confirmed as pregnant with 7 implantations, 5 of which were early resorptions, 1 late resorption and 1 aborted implantation. However, according to the study director, this single abortion is considered not to be related to treatment.
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not examined
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not examined
Changes in number of pregnant:
effects observed, non-treatment-related
Description (incidence and severity):
2/22 females of the control group and of the highest dose group, respectively, 1/22 females of the low-dose group and 3/22 females of the mid-dose group were not pregnant.
Key result
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
other: No adverse maternal toxicity was observed at the highest tested dose level.
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
500 mg/kg bw/d: Fetal weights were marginally lower than the fetal weight of the control group.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not examined
Reduction in number of live offspring:
not examined
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
effects observed, non-treatment-related
Description (incidence and severity):
500 mg/kg bw/d: Litter weights were marginally lower than the fetal weight of the control group.
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
effects observed, non-treatment-related
Description (incidence and severity):
500 mg/kg bw/d: There were two minor fetal pathology findings including a slightly high incidence of additional cranial sutures, and a slightly high incidence of delayed ossification of the cervical vertebrae. According to the study director, the slightly high incidence of additional cranial sutures is in isolation considered not to represent an adverse effect of treatment as cranial bone development was normal. The delayed ossification of the cervical vertebrae represents the results of an evaluation at a snapshot in time (ossification would be expected to continue as the animals matured) and may be linked to the marginally lower mean fetal weights. The incidence of 7th costal cartilage not connected to sternum was lower compared to concurrent controls. According to the study director, the values were slightly high in the control and low-dose group, respectively, and slightly low in the high-dose group when compared with historical control data, thus the finding was considered to be not toxicologically significant.
Visceral malformations:
no effects observed
Other effects:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse embryo-fetal toxicity was observed at the highest tested dose level.
Key result
Abnormalities:
effects observed, non-treatment-related
Localisation:
skeletal: skull sutures
skeletal: vertebra
Description (incidence and severity):
Two minor fetal pathology findings including a slightly high incidence of additional cranial sutures, and a slightly high incidence of delayed ossification of the cervical vertebrae were observed at 500 mg/kg bw/d. According to the study director, the slightly high incidence of additional cranial sutures is considered not to represent an adverse effect of treatment as cranial bone development was normal. The delayed ossification of the cervical vertebrae represents the results of an evaluation at a snapshot in time (ossification would be expected to continue as the animals matured) and may be linked to the marginally lower mean fetal weights.
Key result
Developmental effects observed:
no
Conclusions:
Under the conditions of the study the test substance did not induce any treatment-related biologically relevant malformations in the developing unborn organism in the presence of maternal toxicity. Therefore, the test substance does not meet the criteria for classification for prenatal developmental toxicity according to Regulation (EC) No 1272/2008. The available data is thus conclusive but not sufficient for classification.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
250 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The available information comprises an adequate and reliable study performed with sodium N-lauroylsarcosinate (CAS 137-16-6)
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Rat

The test item sodium N-lauroylsarcosinate (CAS 137-16-6) was administered by gavage to three groups of time-mated pregnant Sprague-Dawley rats, between Days 5 and 19 of gestation, at dose levels of 30, 100, and 250 mg/kg bw/day. A further group of time-mated pregnant females was exposed to the vehicle only (control).

Clinical signs, body weight change, food and water consumptions were monitored during the duration of the study.

At sacrifice (at Day 20 of gestation) all females were subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weight, foetal weight, sex and external and internal macroscopic appearance were recorded. Half of each litter were examined for detailed skeletal development and the remaining half were subjected to detailed visceral examination.

With regard to maternal toxicity one female treated with 250 mg/kg bw/day was found dead on day 10 of gestation. Another female from this treatment group was found dead on Day 18 of gestation. There were no further unscheduled deaths.

Surviving females treated with 250 mg/kg bw/day showed incidences of increased salivation and noisy respiration during the treatment period. One female treated with 100 mg/kg bw/day also showed increased salivation on Day 18 of gestation and a further female showed noisy respiration on Day 13. No such effects were detected in females treated with 30 mg/kg bw/day.

The female found dead on Day 10 did not show any clinical signs prior to Day 10. The female found dead on Day 18 had noisy respiration, increased salivation and pilo-erection.

Females treated with 250 mg/kg bw/day and to a lesser extent females treated with 100 mg/kg bw/day showed a reduction in body weight development throughout the treatment period. Body weight gain adjusted for gravid uterus weight between Days 5 and 20 was also redued in these females. No such effects were detected in females treated with 30 mg/kg bw/day.

Correspondingly, females treated with 250 mg/kg bw/day also showed a reduction in food consumption throughout the treatment period. No such effects were detected in females treated with 100 or 30 mg/kg bw/day.

With regard to developmental toxicity no toxicologically significant effects were detected in the uterine parameters examined, in foetal viability or in growth and development. In addition, no treatment-related effects were detected on foetal external findings. No treatment-related effects were detected on skeletal development or in the type and incidence of skeletal or visceral findings in foetuses from females treated with 250, 100 or 30 mg/kg bw/day. Furthermore there were no findings considered to represent any known malformations.

In conclusion, as the oral administration of the test item to pregnant rats by gavage during gestation at dose levels of 30, 100 and 250 mg/kg bw/day resulted in substance-related effects in females treated with 250 and 100 mg/kg bw/day, the NOAEL for maternal toxicity was therefore considered to be 30 mg/kg bw/day.

No toxicological significant changes were detected in the offspring parameters measured. The NOAEL for reproductive and developmental toxicity was therefore considered to be 250 mg/kg bw/day.

Rabbit

Three groups of 22 females received the test item Sodium N-lauroylsarcosinate (CAS 137-16-6) at doses of 125, 250 or 500 mg/kg/day by oral gavage administration, from Day 6 to 28 after mating. A similarly constituted control group received the vehicle, purified water at the same volume dose as treated groups. Animals were killed on Day 29 after mating for reproductive assessment and fetal examination. Clinical observations, body weight and food consumption were recorded. Adult females were examined macroscopically at necropsy on Day 29 after mating and the gravid uterus weight recorded. All fetuses were examined macroscopically at necropsy and subsequently by detailed internal visceral examination of the head or skeletal examination.

With regard to maternal toxicity one female of the control group and one female each of the mid- and high-dose group died during the study period. However, the deaths were not considered to be treatment-related, since no treatment-related clinical signs were observed after treatment with the test substance at 125, 250 or 500 mg/kg bw/d. In addition, there was no clear effect of treatment on the body weight or food consumption performance of females receiving Sodium N-lauroylsarcosinate (CAS 137-16-6) at 125 or 250 mg/kg bw/d when compared with that of the control group. The mean body weight loss of females exposed to 500 mg/kg bw/d was slightly greater than that of the control group after the start of treatment (between Days 6 and 8 after mating), and body weight gain was slightly lower than that of the controls from Days 8-29 after mating. The food consumption of females treated with 500 mg/kg bw/d was slightly lower than that of the Control from the start of treatment (Day 6 after mating), resulting in a lower overall (Day 6 - 28 after mating) food consumption.

When mean values of body weight and body weight gain were adjusted for the contributions of the gravid uterus, overall maternal mean body weight loss during Days 6-29 of gestation was greater than in controls at 500 mg/kg bw/d. There were no test item-related macroscopic abnormalities detected among the females at scheduled termination on Day 29 after mating.

Embryo-fetal survival was unaffected by treatment at 125, 250 or 500 mg/kg bw/d with mean numbers of implantations, resorptions, live young and percentages of sex ratio and pre and post-implantation loss being similar to Control values.

Mean placental, litter and fetal weights at 500 mg/kg bw/d were marginally lower (ca 10%) than controls.

Mean placental, litter, and male, female and overall fetal weights at 125 or 250 mg/kg bw/day were similar to the control group and unaffected by treatment.

At 500 mg/kg bw/d there was a slightly higher incidence of the minor fetal abnormality additional cranial sutures, which was outside Historical Control Data (HCD).An increase was also seen in the incidence of 20 thoracolumbar vertebrae, however this was within HCD.At 500 mg/kg bw/d there was also a slightly higher incidence of delayed ossification of the cervical vertebrae but this was considered to be related to the lower mean fetal weights seen in this group.

In conclusion, at 500 mg/kg bw/d, maternal body weight performance and food consumption and mean fetal weights were slightly reduced. Embryo-fetal survival was unaffected by treatment and fetal development was not adversely affected. The No-Observed-Adverse-Effect-Level (NOAEL) for maternal and embryo-fetal toxicity was concluded to be 500 mg/kg bw/d.

Justification for classification or non-classification

The available information on reproductive and developmental toxicity of sodium N-lauroylsarcosinate according to the criteria laid down in regulation (EC) No 1907/2006, Annex IX, 8.7.2, does not meet the criteria for classification according to Regulation (EC) No 1272/2008. The available data is thus conclusive but not sufficient for classification.