Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose for cross-reference:
read-across source
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
other information
Justification for type of information:
Justification is provided in the separate statement.
Reason / purpose for cross-reference:
assessment report
Conclusions:
The reproduction toxicity the registration substance sodium N-lauroyl glutamate is derived based on the read-across sodium N-cocoyl glycinate. No significant potential for reproduction toxicity can be reliably derived.
Executive summary:

The reproduction toxicity of the registration substance "sodium N-lauroyl glutamte" was evaluated based on the read-across approach using the reproduction toxicity data for sodium N-cocoyl glycinate.

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compound are likely exhibit comparable toxicity profiles.

The reproduction toxicity of sodium cocoyl glycinate was investigated according to the Guideline OECD 421. The test material, composed of 70% sodium cocoyl glycinate and 20 % sodium chloride, was dissolved in water and was administered to rats via gavage at dosages of 0, 62.5, 250, and 1000 mg/kg bw/day, corresponding to up to 700 mg/kg bw/day of sodium cocoyl glycinate. It was given to male rats for 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the offsprings reached day 4 post partum.

At high dose, the body weight and the food consumption of males and females were reduced. These effects were considered as not adverse nature for parental animals, because the relative difference to the control values were within 10% and seemed to stagnate with the treatment progression. No other effect was noted for parental animals.

With respecto the reproduction performance no effects on mating performance, fertility, corpora lutea count or duration of gestation were observed at any dose level. At high dose, the numbers of alive pups at first check and on lactation day 4 were reduced. This effect was considered as consequence of reduced food uptake and/or high salt content of the test material and therefore not considered as evidence of specific reproduction toxicity of sodium cocoyl glycinate.

The NOAEL of 250 mg/kg bw/day was established for reproduction toxicity of the test material.

Likewise, the registration substance "sodium N-lauroyl glutamate" is considered as ot low reproduction toxicity.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Reference substance name:
Sodium cocoyl glycinate (SCG) [INCI]
IUPAC Name:
Sodium cocoyl glycinate (SCG) [INCI]
Details on test material:
Stability of Test Item: Stable under storage conditions
Stability of Test Item Dilution: Unknown in PEG 300; excluded from the statement of compliance.
Storage Conditions: At room temperature (range of 20 ± 5 °C, provided by Harlan Laboratories Ltd.), light protected.
Safety Precautions: Routine hygienic procedures were used to ensure the health and safety of the personnel.
Description: Colorless solid
Specific details on test material used for the study:
Composition of the test material:
68.1 % N-cocoyl glycine sodium salt
19.6% Sodium chloride
4.4 % Glycine
6.9% Fatty acid

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories BV
- Age at study initiation: approx. 7 weeks
- Weight at study initiation: 199 - 233 g (males), 133 - 154 g (females)
- Fasting period before study: yes (overnight)
- Housing: in groups of 5 in Makrolon type-4 cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 10 - 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hour dark / light cycle

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): n.a.
- Mixing appropriate amounts with (Type of food): n.a.
- Storage temperature of food: n.a.

VEHICLE
- Justification for use and choice of vehicle (if other than water): purified water
- Concentration in vehicle: 0 - 100 mg/mL
- Amount of vehicle (if gavage): 10 mL (dose volume)
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: until evidence of copulation (up to 14 days pairing period)
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
- After 14 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: individually
- Any other deviations from standard protocol: no
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Males: minimum 4 weeks
Females: approximately 7 weeks
Frequency of treatment:
once daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
62.5 mg/kg bw/day
Dose / conc.:
250 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
11 per sex per group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on dose-range-finding study
- Rationale for animal assignment: random

Examinations

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

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: daily from start of treatment to day of necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): n.a.


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): n.a.

Sperm parameters (parental animals):
Parameters examined in [all/P/F1/F2] male parental generations:
No abnormal microscopic findings during sperm staging regarding completeness of stages and maturation of cell populations.
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was determined for pups born or found dead
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals after treatment for 28 days.
- Maternal animals: All surviving animals on day 21 post coitum (if birth did not occur at that day, the dam was sacrificed on day 25 post coitum).

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
Postmortem examinations (offspring):
SACRIFICE
The F1 offspring were sacrificed at 4 days of age.

GROSS NECROPSY
No test item related findings in any pubs at any dose level observed.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
At the dose level of 1000 mg/kg bw/day, bedding in mouth was observed in all males starting from day 5 of the treatment and in all females starting from day 8 of the treatment until the completion of the treatment.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Males:
At the dose level of 1000 mg/kg bw/day, statistically significant reduction in body weights and body weight gains were noted in males. Reduction in body weights was noted from day 5 of the pre-pairing period until the completion of the study, reduction in body weight gain was noted from day 5 to 14 of the pre-pairing period. During pairing period, body weight gain of males at the high dose level was similar to the body weight gain in the control group.
Females:
At the dose level of 1000 mg/kg bw/day, lower body weights (not statistically significantly) and lower body weight gain (statistically significant on individual days) were noted during gestation.

The effect found for food consumption and body weight were considered as treatment related but not adverse, because the relative differecen to terminal body were not severe (less than 10%) and seemed to stagnate with the progression of the treatment.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
For males in pre-pairing Period:
At the dose level of 1000 mg/kg bw/day, statistically significant reduction in food consumption was noted in males. Mean food consumption over the pre-pairing period was 21.2 g/animal/day compared to 25.2 g/animal/day in the control group.
For females in pre-pairing, gestation and lactation periods:
At the dose level of 1000 mg/kg bw/day, statistically significant reduction in food consumption was noted in females during the pre-pairing and gestation periods. During the lactation period, food consumption remained lower but not statistically significantly. Mean food consumption during the pre-pairing, gestation and lactation periods was at the high dose level: 16.0, 19.8 and 21.7 g/animal/day, compared to the respective values of 18.3, 22.9 and 25.0 g/ animal/day in the
control group.

The effect found for food consumption and body weight were considered as treatment related but not adverse, because the relative differecen to terminal body were not severe (less than 10%) and seemed to stagnate with the progression of the treatment.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
At dose of 1000 mg/kg/day the number of pups born alive and the number of pups on LD4 were reduced.

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
250 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
reproductive performance
Remarks on result:
other:
Remarks:
The findings of reduced number of alive pups at first check and on lactation day 4 are not considered as evidence of reproduction toxicity of sodium cocoyl glycinate. - At dose of 1000 mg/kg/day, the body weight and the food consumption of males and females were reduced. These effects were considered as not adverse nature for parental animals, because the relative difference of body weight to the control values were within 10% and seemed to stagnate with the treatment progression. Nevertheless, it is reasonable to consider the reduced viability of offspings as the consequence of reduced food uptake during pregnancy and laction of dams. - The test material contained 20% NaCl and the animals were in fact treated with NaCl up to 200 mg/kg/day. It is well-known that high salt maternal intake is associated with adverse effects on the offspring (i.e. Maruyama K et al., Lif Sci.2015 Sep 1; 136:42-51). The reduced viability of offsprings could be related to the high salt intake of dams as well. In conclusion, the effects found at 1000 mg/kg/day is not to be interpreted as the evidence of reproduction toxicity of sodium cocoyl glycinate.

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
At 1000 mg/kg bw/day, the numbers of pups alive at first check and on LD4 were reduced.
Body weight and weight changes:
no effects observed

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
250 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
mortality
Remarks on result:
other:
Remarks:
The findings of reduced number of alive pups at first check and on lactation day 4 are not considered as evidence of reproduction toxicity of sodium cocoyl glycinate. - At dose of 1000 mg/kg/day, the body weight and the food consumption of males and females were reduced. These effects were considered as not adverse nature for parental animals, because the relative difference of body weight to the control values were within 10% and seemed to stagnate with the treatment progression. Nevertheless, it is reasonable to consider the reduced viability of offspings as the consequence of reduced food uptake during pregnancy and laction of dams. - The test material contained 20% NaCl and the animals were in fact treated with NaCl up to 200 mg/kg/day. It is well-known that high salt maternal intake is associated with adverse effects on the offspring (i.e. Maruyama K et al., Lif Sci.2015 Sep 1; 136:42-51). The reduced viability of offsprings could be related to the high salt intake of dams as well. In conclusion, the effects found at 1000 mg/kg/day is not to be interpreted as the evidence of reproduction toxicity of sodium cocoyl glycinate.

Overall reproductive toxicity

Key result
Reproductive effects observed:
yes
Lowest effective dose / conc.:
1 000 mg/kg bw/day
Treatment related:
yes
Relation to other toxic effects:
reproductive effects as a secondary non-specific consequence of other toxic effects
Dose response relationship:
yes
Relevant for humans:
no

Any other information on results incl. tables

Table 1: Summary of Food Consumption of Males

Doses

[mg/kg /day]

 

Food Consumption of Males [g]; n = 11

Pre-paring

Day 1-8

Pre-paring

Day 8-14

0

Mean

24.9

25.5

S.D.

1.4

1.5

62.5

Mean

24.7

24.8

S.D.

2.4

2.3

250

Mean

24.2

23.8

S.D.

2.4

2.5

1000

Mean

20.7 **

21.7 **

S.D.

3.7

3.4

*/**: DUNNETT-Test based on pooled variance sig. at 5% (*), 1% (**)

Table2: Summary of Food Consumption of Females; Prepairing, Gestation and Lactation

Doses

[mg/kg /day]

 

Food Consumption of Females [g]

Pre-pairing

 

Gestation

 

Lactation

Day 1-8

Day 8-14

GD 0-7

GD 7-14

GD 14-21

LD 1-4

0

Mean

18.1

18.4

22.8

23.5

22.4

25.0

S.D.

1.3

1.6

1.7

1.9

1.4

6.7

n

11

11

8

8

8

9

62.5

 

Mean

17.8

18.2

21.8

22.4

21.1

26.2

S.D.

1.3

1.1

1.2

0.7

1.0

4.4

n

11

11

10

10

10

11

250

Mean

18.4

18.7

22.3

23.1

21.5

26.9

S.D.

2.7

1.9

2.2

2.2

2.4

3.4

n

11

11

11

11

11

11

1000

Mean

15.6 *

16.4 *

19.6 **

20.0 **

19.8 *

21.7

S.D.

2.0

1.7

2.2

2.9

2.2

5.6

n

11

11

10

10

10

10

*/**: DUNNETT-Test based on pooled variance sig. at 5% (*), 1% (**)

Table 3: Summary of Body Weights of Males

Doses

[mg/kg /day]

 

Body weight of males [g]; n = 11

Pre-pairing period

Pairing period

1d

8d

14d

1d

8d

15d

0

Mean

311

335

354

350

367

381

S.D.

8

12

12

12

13

13

62.5

Mean

310

332

349

348

362

379

S.D.

8

7

11

13

13

12

250

Mean

310

330

345

343

360

376

S.D.

8

12

15

14

18

21

1000

Mean

310

318**

332**

328**

346*

363*

S.D.

8

17

19

19

18

18

*/**: DUNNETT-Test based on pooled variance sig. at 5% (*), 1% (**)

Table 4: Summary of Body Weights of Females; Pre-mating

Doses

[mg/kg /day]

 

Body weight [g]; n = 11

1d

8d

14d

0

Mean

195

203

209

S.D.

8

10

10

62.5

Mean

194

204

211

S.D.

6

8

11

250

Mean

196

205

210

S.D.

6

7

8

1000

Mean

197

201

207

S.D.

8

11

11

*/**: DUNNETT-Test based on pooled variance sig. at 5% (*), 1% (**)

Table 5: Summary of Body Weights of Females; Gestation and Lactation

Doses

[mg/kg /day]

 

Body weight [g]

GD0

GD7

GD14

GD21

 

LD1

LD4

0

Mean

211

240

268

345

 

240

252

S.D.

11

16

15

19

 

14

17

n

8

8

8

8

 

9

9

62.5

 

Mean

211

240

268

337

 

244

256

S.D.

11

12

13

14

 

15

13

n

10

10

10

10

 

11

11

250

Mean

216

244

269

334

 

245

258

S.D.

11

10

13

22

 

16

10

n

11

11

11

11

 

11

11

1000

Mean

214

234

261

329

 

238

251

S.D.

15

14

16

16

 

18

15

n

10

10

10

10

 

10

10

One female group 1 and one female group 2: mating not detected, therefore body weight for gestation missing

Table 6. Summary of reproductive parameters

Dose (mg/kg bw)

0

62.5 mg/kg/day

250 mg/kg/day

1000 mg/kg/day

 

 

 

 

 

Number of females paired

 

11

11

11

11

Number of females mated (confirmed by vaginal smear)

10

10

11

11

Number of females pregnant

(confirmed at littering/necropsy)

9

11

11

10

Dystocia death

0

0

0

0

Number of females with live litters

9

11

11

10

Precoital interval (days)

2.4 ± 1.3 (n = 10)

2.4 ± 1.0 (n = 10)

4.3 ± 3.3 (n = 11)

4.7 ± 4.0 (n = 11)

Gestation length (days)

21.6± 0.5(n= 8)

21.3± 0.5(n=10)

21.5 ± 0.5(n= 11)

21.6 ± 0.5(n = 10)

Corpora lutea

14.1 ± 1.2 (n= 9)

14.0 ± 1.3 (n=11)

14.3 ± 1.4 (n= 11)

14.2± 1.9 (n=10)

Implantation sites

13.8 ± 1.3 (n= 9)

13.0 ± 1.1 (n=11)

13.1 ± 3.2 (n= 11)

13.3 ± 2.3 (n=10)

Post implantation loss

0.6 ± 0.7(n= 9)

0.7 ± 1.1(n= 11)

1.5 ± 2.1(n= 11)

2.7 ± 2.8(n= 10)

Viable litter size at first check

13.2 ± 1.4(n= 9)

12.3 ± 1.6(n= 11)

11.6 ± 3.7(n= 11)

10.6 ± 2.0(n= 10) *

Dead pups at first check

0.0

0.1 ± 0.3(n= 11)a

0.0

0.6 ± 1.9(n= 10)b

Viable litter size LD4

13.2 ± 1.4(n= 9)

11.9 ± 1.8(n= 11)

11.5 ± 3.6(n= 11)

9.4 ± 3.3(n= 10) *

 

 

 

 

 

Total number born alive pups at first check

119

135

128

106

% of males/females of born alive pups at first check

54/46

49/51

50/50

42/58

Total number alive pups on LD 4

119

131

127

94

 

 

 

 

 

Body weight of pups (g) on LD1

5.9 ± 0.7 (n = 9)

5.7 ± 0.6 (n = 11)

6.0 ± 0.6 (n= 11)

5.7 ± 0.9 (n = 10)

Body weight of pups (g) on LD4

8.1 ± 1.4 (n = 9)

8.6 ± 1.0 (n = 11)

8.8 ± 1.3 (n = 11)

8.2 ± 1.5 (n = 10)

 

 

 

 

 

One female group 1 and one female group 2: mating not detected, therefore gestation length missing

*/**: Steel Test, significant at 5% (+), 1% (++)

a) One pup found dead.

b) Six pups found dead, whereas only one litter (animal no. 83) was affected. Additionally eight alive pups were found at first check for this litter.

Applicant's summary and conclusion

Conclusions:
The reproduction toxicity of sodium N-cocoyl glycinate was investigated according to the Guideline OECD 421. No significant potential of reproducton toxicity was found for sodium N-cocoyl glycinate.
Executive summary:

The reproduction toxicity of sodium cocoyl glycinate was investigated according to the Guideline OECD 421. The test material, composed of 70% sodium cocoyl glycinate and 20 % sodium chloride, was dissolved in water and was administered to rats via gavage at dosages of 0, 62.5, 250, and 1000 mg/kg bw/day, corresponding to up to 700 mg/kg bw/day of sodium cocoyl glycinate. It was given to male rats for 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the offsprings reached day 4 post partum.

At high dose, the body weight and the food consumption of males and females were reduced. These effects were considered as not adverse nature for parental animals, because the relative difference to the control values were within 10% and seemed to stagnate with the treatment progression. No other effect was noted for parental animals.

With respecto the reproduction performance no effects on mating performance, fertility, corpora lutea count or duration of gestation were observed at any dose level. At high dose, the numbers of alive pups at first check and on lactation day 4 were reduced. This effect was considered as consequence of reduced food uptake and/or high salt content of the test material and therefore not considered as evidence of specific reproduction toxicity of sodium cocoyl glycinate.

The NOAEL of 250 mg/kg bw/day was established for reproduction toxicity of the test material.