Fatty acid chlorides, C8-14 (even numbered), reaction products with glycine

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

The registration substance Hostapon SG (70% Sodium N-cocoyl glycinate) was investigated for its reproduction toxicity 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. 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 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 respect to 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. It is well-known that high salt maternal intake is associated with adverse health effects(i.e. reviewed by EPA 2003: "Drinking Water Advisory: Consumer Acceptability Advice and Health Effects Analysis on Sodium).The reduction of alive pups is considered as consequence of reduced food uptake and high salt content of the test material and therefore not considered as evidence of specific reproduction toxicity of Sodium N-cocoyl glycinate.

The NOAEL of 250 mg/kg bw/day was established for reproduction toxicity of Hostapon SG.

Link to relevant study records

Reference

Reference 1

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

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

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

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

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

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.

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: 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.

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.

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

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.

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

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.

Conclusions:

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

Executive summary:

The registration substance Hostapon SG (70% Sodium N-cocoyl glycinate) was investigated for its reproduction toxicity 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. 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 respect to 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. It is well-known that high salt maternal intake is associated with adverse health effects (i.e. reviewed by EPA 2003: "Drinking Water Advisory: Consumer Acceptability Advice and Health Effects Analysis on Sodium). The reduction of alive pups is considered as consequence of reduced food uptake and high salt content of the test material and therefore not considered as evidence of specific reproduction toxicity of Sodium N-cocoyl glycinate.

The NOAEL of 250 mg/kg bw/day was established for reproduction toxicity of Hostapon SG.

Reference 2

Endpoint:

extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the extended one-generation reproductive toxicity study does not need to be conducted because there are no results from available repeated dose toxicity studies that indicate adverse effects on reproductive organs or tissues, or reveal other concerns in relation with reproductive toxicity

Effect on fertility: 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:

One valid guideline study

Additional information

.

Effects on developmental toxicity

Description of key information

The developmental toxicity of the registration substance Hostapon SG was assesses via read-across to Sodium N-Lauroyl-Sarcosinate.

The developmental toxicity of Sodium N-Lauroyl-Sarcosinate in rats was investigated according to the Guideline OECD 414. Time mated pregnant rats were treated at doses of 0, 30, 100 and 250 mg/kg/day. Reduced food consumption at 250 mg/kg/day and reduced body weight gain was noted at doses of 100 and 250 mg/kg/day. Sloughing on the non-glandular region of the stomach was found at 250 mg/kg/day. No effect was found for fetuses. NOAEL of 250 mg/kg/day was determined for overall developmental toxicity.

Likewise, Hostapon SG is expected to be of no developmental toxicity.

Link to relevant study records

Reference

Reference 1

Endpoint:

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 Chapter 13.

Reason / purpose for cross-reference:

assessment report

Conclusions:

The developmental toxicity the registration substance Hostapon SG is derived based on the read-across to sodium N-lauroyl sarcosinate. Hostapon is expected to be of no developmental toxicity.

Executive summary:

The developmental toxicity of the registration substance Hostapon SG was assesses via read-across to Sodium N-Lauroyl-Sarcosinate.

The developmental toxicity of Sodium N-Lauroyl-Sarcosinate in rats was investigated according to the Guideline OECD 414. Time mated pregnant rats were treated at doses of 0, 30, 100 and 250 mg/kg/day. Reduced food consumption at 250 mg/kg/day and reduced body weight gain was noted at doses of 100 and 250 mg/kg/day. Sloughing on the non-glandular region of the stomach was found at 250 mg/kg/day. No effect was found for fetuses. NOAEL of 250 mg/kg/day was determined for overall developmental toxicity.

Likewise, Hostapon SG is expected to be of no developmental toxicity.

Reference 2

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

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

adopted 22 January 2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)

Version / remarks:

adopted August 1998

Deviations:

no

Qualifier:

according to guideline

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. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD® (SD)IGS BR

Details on test animals or test system 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.

Key result

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

Key result

Developmental effects observed:

no

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:

Sodium N-Lauroyl-sarcosinate was investigated according to the Guideline OECD 414. NOAEL of 250 mg/kg/day was determined.

Executive summary:

The developmental toxicity of the registration substance Hostapon SG was assesses via read-across to Sodium N-Lauroyl-sarcosinate.

The developmental toxicity of Sodium N-Lauroyl-sarcosinate in rats was investigated according to the Guideline OECD 414. Time mated pregnant rats were treated at doses of 0, 30, 100 and 250 mg/kg/day. Reduced food consumption at 250 mg/kg/day and reduced body weight gain was noted at doses of 100 and 250 mg/kg/day. Sloughing on the non-glandular region of the stomach was found at 250 mg/kg/day. No effect was found for fetuses. NOAEL of 250 mg/kg/day was determined for overall developmental toxicity.

Likewise, Hostapon SG is expected to be of no developmental toxicity.

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:

One valid guideline study on the read-across source substance

Justification for classification or non-classification

No indication of reproduction toxicity was found in the reproduction toxicity screening test on the registration substance and in the developmental toxicity study on the read-across source chemical. No classification is assigned to the registration substance.