Reaction mass of sodium 1-methoxy-1-oxohexadecane-2-sulphonate and sodium methyl 2-sulphooctadecanoate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Data from OECD test guideline study and GLP principles on the main component of the reaction mass:

C16MES: NOAEL >= 300 mg/kg bw/day for reproductive performance of parental animals and for offspring development

Data on similar substances:

C14MES: NOAEL >= 740 mg/kg bw/day for male rats and > 1039 mg/kg bw/day for female rats for the reproductive and developmental toxicity

Link to relevant study records

Reference

Reference 1

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

The study has been performed according to OECD guidelines. No data was present in the publication on whether the study was performed according to GLP principles. All data related to the reproduction study was not included in the publication, so the results cannot be assessed.

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

not specified

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

not specified

Details on mating procedure:

During the mating period, each male was placed in the cage of his paired female overnight until pregnancy occurred. The females rats were examined each morning for the presence of sperm and the day it was found was stated as day 0.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Males: 56 days including 14 days prior to mating, during the mating and post-mating periods
Females: 41-45 days (from 14 days prior to mating until post-partum Day 4)

Frequency of treatment:

Contineously

Dose / conc.:

0.3 other: % in diet (nominal)

Remarks:

175 and 249 mg/kg bw/day for males and females, respectively

Dose / conc.:

0.6 other: % in diet (nominal)

Remarks:

360 and 497 mg/kg bw/day for males and females, respectively

Dose / conc.:

1.2 other: % in diet (nominal)

Remarks:

740 and 1039 mg/kg bw/day for males and females, respectively

No. of animals per sex per dose:

5

Control animals:

yes, plain diet

Dose descriptor:

NOEL

Effect level:

> 740 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: reproductive toxicity

Dose descriptor:

NOEL

Effect level:

> 1 039 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: reproductive toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

> 740 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: reproductive toxicity

Critical effects observed:

not specified

Remarks on result:

not measured/tested

Critical effects observed:

not specified

Dose descriptor:

NOEL

Generation:

F1

Effect level:

> 1 039 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: developmental toxicity

Dose descriptor:

NOEL

Generation:

F1

Effect level:

> 1 039 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: developmental toxicity

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

> 1 039 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: developmental toxicity

Critical effects observed:

not specified

Remarks on result:

not measured/tested

Critical effects observed:

not specified

Reproductive effects observed:

not specified

The reproduction/developmental parameters were essentially the same in all treatment groups and the control.

These absence of adverse effects in the C14 MES dosed groups indicate that the NOAEL for the reproductive and developmental toxicity was more than 1.2% of the diet in both sexes.

Conclusions:

These absence of adverse effects in the C14 MES dosed groups indicate that the NOAEL for the reproductive and developmental toxicity was more than 1.2% of the diet in both sexes.

Reference 2

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 2, 2000 to May 29, 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- strain: Crj: CD (SD) IGS
- Source:Charles River Laboratories Japan, Inc
- Age at study initiation: 8 weeks
- Weight at study initiation: 364 to 420 g for males and 223 to 264 g for females.
- Fasting period before study: no
- Housing: in individual stainless steel cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): >= 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Administered suspensions were prepared by mixing the substance with Japanese Pharmacopoeia-grade olive oil to designated concentration doses.

Details on mating procedure:

Estrous cycle test:
estrous cycle stages were determined microscopically

Observation of mating and delivery:
- on Day 15 of administration. Each pair was housed together in the same cage until mating was confirmed, with two weeks as the limit. The mating rate (%) [(number of animals that had mated/number of animals housed together) x 100] was calculated. Mating was confirmed with formation of the copulatory plug or sperm in vaginal smears (day 0 of pregnancy). Observation of the delivery state was also carried out at the same time, and the day upon which completion of delivery was confirmed was designated as Day 0 of lactation. Based on the observation results of mating and delivery, the conception rate (%) [(number of females conceiving/number of females that had mated) x 100] and birth rate (%) [(number of females giving birth/number of pregnant females) x 100] were calculated.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The hexadecanoic acid, 2-sulfo-, 1-methylester, sodium salt suspensions of 0.1 w/v% (for 5 mg/kg group), 0.4 w/v% (for 20 mg/kg group), 1.6 w/v% (for 80 mg/kg group) and 6.0 w/v% (for 300 mg/kg) prepared for the first and last times were analyzed using the HPLC method. The analysis was repeated three times, and the mean values were calculated.

Date of preparation Displayed value (w/v %)
0.1 0.4 1.6 6.0

May 30, 2000 0.10 0.40 1.7 6.2
July 8, 2000 0.09 0.40 1.6 5.8

The results shown below confirm that the suspensions were prepared at the designated concentrations.

Duration of treatment / exposure:

The administration duration ranged from 14 days before start of mating for both males and females, lasting 47 days for males and to 42 days at the shortest to 55 days at longest after birth just before day 4 of lactation for females.

Frequency of treatment:

once a day

Details on study schedule:

Premating exposure period for male and female: 14 days

Dose / conc.:

5 mg/kg bw/day (actual dose received)

Dose / conc.:

20 mg/kg bw/day (actual dose received)

Dose / conc.:

80 mg/kg bw/day (actual dose received)

Dose / conc.:

300 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Positive control:

Not applicable.

Parental animals: Observations and examinations:

See Section 7.5.1 RA 223-676-0 (OECD 422)

Oestrous cyclicity (parental animals):

Estrous cycle
No. of pairs with successful copulation
No. of pregnant females
No. of pregnant females with live pups gestation lenght
No. of corpora lutea
No. of implantation sites

Sperm parameters (parental animals):

Parameters examined in all male parental generations: testis weight

Litter observations:

No. of pups born
No. of pups alive on day 0 of lactation
Sex ratio
No. of dead pups

Postmortem examinations (parental animals):

See Section 7.5.1 RA 223-676-0 (OECD 422)

Postmortem examinations (offspring):

The main organs in the chest and abdomen were observed macroscopically in dead pups at the time of death and in live pups, which were exsanguinated under ether anesthetic on Day 4 of lactation.

Statistics:

See Section 7.5.1 RA 223-676-0 (OECD 422)

Reproductive indices:

Copulation index (No. pairs with successful copulation/No. of pairs mated x 100)
fertility index (No. of pregnant animals/No. of animals with successful copulation x 100)
gestation index (No. of females with live pups/No. of pregnant females x 100)
implantation index (No. of implants/No. of corpora lutea x 100)
delivery index (No. of pups born/No. of implants x 100)

Offspring viability indices:

live birth index (No. of live pups born/No. of pups born x 100)
viability index on day 4 (No. of live pups on day 4 after birth/No. of live pups born x 100)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

See section 7.5.1 RA 223-676-0 (OECD 422)

Estrous cycle examination: No significant differences in the estrous cycle were noted between the study groups and the control group.

Mating and conception rates: All pairs successfully mated except for one in the 20 mg/kg group. No significant difference was seen in the time taken for mating. Conception in the control group was 70% because of failure in three pairs whose males exhibited abnormality of spermatogenesis, while the parameter was 100% for all study groups.

Corpora lutea count, number of implantations, and implantation rate:
No significant differences in corpora lutea count, the number of implantations, or implantation rate were found between any of the study groups and the control group.

Key result

Dose descriptor:

NOAEL

Effect level:

>= 300 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive performance

Critical effects observed:

not specified

Remarks on result:

not measured/tested

Critical effects observed:

not specified

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

No effects of the substance were observed on the viability and body weight of offspring. No malformations were found in offspring in any groups.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 300 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: for offspring development, no effects observed

Remarks on result:

not measured/tested

Critical effects observed:

not specified

Reproductive effects observed:

not specified

Reproductive and developmental parameters: No effects of this substance were observed on reproductive performance in males and females or on viability and body weight of offspring. No malformations were found in offspring in any groups.

Dose (mg/kg bw/day)	0	5	20	80	300
Estrous cycle (days)
Mean	4.2	4.0	4.0	4.0	4.0
SD	0.6	0.0	0.0	0.1	0.1
No. of pairs mated	10	10	10	10	10
No. of pairs copulated	10	10	9	10	10
Copulation index (%)	100	100	90	100	100
No. of pregnant females	7	10	9	10	10
Fertility index (%)	70	100	100	100	100
No. of pregnant females with parturition	7	8	9	10	10
No. of pregnant females with live pups	7	8	9	10	10
Gestation length(days)
Mean	22.4	22.5	22.4	22.6	22.5
SD	0.5	0.5	0.5	0.5	0.5
No. of corpora lutea
Mean	19.7	18.4	19.3	17.5	18.2
SD	2.8	4.1	3.3	1.9	2.6
No. of implantation sites
Mean	15.6	13.2	14.9	15.9	15.3
SD	1.6	6.3	2.4	1.5	2.4
No. of pups born
Mean	15.0	11.9	13.1	14.8	13.9
SD	1.7	6.4	4.0	2.0	1.7
Delivery index(%)
Mean	96.3	73.8	85.4	93.3	91.4
SD	4.8	39.4	20.0	10.1	6.1
Sex ratio(Male/female)	0.78	0.98	0.97	0.85	0.88
No. of pups alive on day 0 of lactation
Mean	15.0	14.9	13.0	14.4	12.3
SD	1.7	1.6	3.9	2.0	4.2
Live birth index(%)
Mean	100	100	99.3	97.4	88.3
SD	0	0	2.1	4.6	27.2
No. of pups alive on day 4 of lactation
Mean	14.7	14.9	12.9	14.2	12.0
SD	1.4	1.6	3.8	1.8	4.7
Viability index
Mean	98.3	100	99.3	98.7	89.3
SD	2.9	0	2.2	2.7	31.5

Conclusions:

No effects of the substance were observed on reproductive performance in males and females or on viability and body weight of offspring. The NOAEL was determined to be 300 mg/kg bw/day for reproductive performance of parental animals and for offspring development.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

300 mg/kg bw/day

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

In an oral reproduction/developmental toxicity screening study, performed according to OECD 422 test guidelines, rats were exposed to 0, 5, 20, 80 and 300 mg/kg bw/d C16MES by gavage. The administration duration ranged from 14 days before start of mating for both males and females, lasting 47 days for males and to 42 days at the shortest to 55 days at longest after birth just before day 4 of lactation for females. Accuracy of formulations was demonstrated by analyses. No treatment related changes were noted in any reproductive parameters investigated in this study (i.e. mating, fertility and conception indices, precoital time, and numbers of corpora lutea and implantation sites). Therefore, the reproduction NOAEL was determined to be >= 300 mg/kg bw/d.

In a supporting oral reproduction/developmental toxicity screening study, performed according to OECD 422 test guidelines, rats were exposed to 0, 0.3, 0.6 and 1.2% C14MES via the diet. Males were exposed for 56 days including 14 days prior to mating, during the mating and post-mating periods, females were exposed for 41-45 days (from 14 days prior to mating until post-partum Day 4). No treatment related changes were noted, therefore, the reproduction NOAEL was determined to be >= 780 mg/kg bw/d (equivalent to 1.2%).

Although no data is available on the component C18MES, it was judged that the NOAEL for fertility of the reaction mass could be based on the data of the main component of the reaction mass and the data of a similar substance with shorter alkyl chain length.

Effects on developmental toxicity

Description of key information

Data from OECD test guideline 414 and GLP principles dermal study with C12/C14/C16 (1/2/7) MES: NOAEL systemic is >=15% (equivalent to 101.3 mg/kg bw/d). The maternal NOAEL for local effects is 5% (equivalent to 33.2 mg/kg bw/day) based on slight irritation at the application site.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The study was not performed according to GLP, but was well described, and basically agreed with OECD 414 (2001), although there were deviations.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

See below.

Qualifier:

according to guideline

Guideline:

other: See below

Deviations:

not specified

Principles of method if other than guideline:

It was reported that the study complied with the following guidelines: Guidelines for Toxicity Studies Required for Applications to Manufacture (Import) Drugs” Notification No. 24 of the Pharmaceutical Affairs Bureau, Ministry of Health and Welfare, September 11, 1989; 1990 Guidelines for Toxicity Studies of Drug Manual, First Edition (published in 1991)., Editorial Supervision by New Drugs Division Pharmaceutical Affairs Bureau, Ministry of Health and Welfare of Japan, Yakuji Nippo Ltd.
The study was performed in agreement with OECD 414 (2001), with the following deviations: the scheme for dose administration (day 7 through 17 of presumed gestation) does not agree with OECD 414 (2001), which states day 5 post mating to the day prior to scheduled caesarian section. In addition, the gravid uterine weight was not determined.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc.
- Age at study initiation: males 9 weeks, females 8 weeks
- Weight at study initiation: upon receipt (day -6) males 273–299 g, females 156–183 g
- Fasting period before study: no
- Housing: Rat cage; Quarantine period: 3–4 animals per cage; Mating period 1 male and 1 female per cage; Test period: 1 animal per cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26
- Humidity (%): 30-70
- Air changes (per hr): >= 15
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: November 5, 1990 To: December 17, 1990

Route of administration:

dermal

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: 3 x 4 centimeter area
- % coverage: the entire 3 x 4 cm area was treated with 0.2 mL of test formulation
- Type of wrap if used: none
- Time intervals for shavings or clipplings: On the day before administration (Day 6 of pregnancy), the hair of animals was clipped using an electric hair clipper to create a 3 x 4 centimeter area for application. Thereafter, hair was clipped several times per week during the administration period, according to hair growth.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.2 mL
- Concentration (if solution): 1.5, 5 and 15 w/v%
- Constant volume or concentration used: yes
- For solids, paste formed: The test substance is solid at an ambient temperature. In order for it to assume the form of a uniform paste, it was melted in a water bath of approximately 70°C, then heated at approximately 40°C to prepare a study solution for administration at a concentration of 15 w/v% (calculated on the assumption of 70.3% purity), using distilled water. This preparation process was performed once a week, and the prepared solution was designated as “the preservation solution.” The preservation solution was stored at ambient temperature. Some of the solution was taken each day and diluted with distilled water to prepare 5- and 1.5-w/v% study solutions for administration. In cases in which the test substance was separated in the preservation solution at the time of preparation, the preparation was heated in warm water at approximately 40°C before it was used as the study solution.

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1 M with 1 F
- Length of cohabitation: overnight
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility: no data.
- Further matings after two unsuccessful attempts: no data
- Verification of same strain and source of both sexes: yes
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no

Duration of treatment / exposure:

from day 7 through 17 of presumed gestation.

Frequency of treatment:

Once daily

Duration of test:

21 days (Caesarian section was performed on day 20 of presumed pregnancy).

Remarks:

Doses / Concentrations:
1.5, 5, 15
Basis:
other: % w/v active ingredient

No. of animals per sex per dose:

There were 27 or 28 rats per group that were confirmed to have mated at the time of group assignment. The number of pregnant animals was 26 in the control group and 25 in each of the treated groups.

Control animals:

yes, concurrent vehicle

Details on study design:

Three sets of doses were chosen for the study, with 15 w/v% solution being set as the maximum concentration (highest dose) based on the results of a 28-day repeated dose dermal toxicity study (study ID: 90007/90077) in which mild skin irritation was observed. The other doses were 5 w/v% and 1.5 w/v% .

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily. Observation of the topical application site was performed daily prior to application. The findings were rated in accordance with the Draize method .
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: Days 0, 3, 5, 7-18 and 20 of pregnancy.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Time schedule for examinations: Days 0, 3, 5, 7-18 and 20 of pregnancy.
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus with ovaries were excised for intrauterine examinations.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: placental weight was determined; the number of placental remnants per uterine horn was determined. Any case of remaining placenta without a fetus (i.e. a case in which placental formation was followed by resorption of fetus, leaving placenta only) was counted as a placental remnant case.

Fetal examinations:

- External examinations: Yes: [all per litter]; fetuses were also weighed and sex was determined
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
- Head examinations: Yes, external examination

Statistics:

Animals that showed no sign of pregnancy during the test period and were confirmed not to be pregnant on Day 20 or those that gave birth before the scheduled cesarean operation on Day 20 were excluded from the analysis. Measurements obtained in this study were first checked by Bartlett test for homoscedasticity at a 1% level of significance. If they were found to be homoscedastic, multiple comparisons were performed using Dunnett's test between the control group and each study substance group; if there was no homoscedasticity, attribution to an outlier was checked. If an outlier was excluded from statistical analysis, results were dismissed and the homoscedasticity check was performed again, followed by multiple comparisons between the control group and each of the study substance groups using Dunnett's test. If an outlier could not be excluded from the statistical analysis, Dunnett's test was still performed for multiple comparisons between the control group and each of the study substance groups.
For rates of occurrence of anomalies and alternations in fetuses, rates of implantations, and embryonic/fetal mortality rates, multiple comparisons were performed between the control group and each of study substance groups using Steel's test.
To examine the sex ratio, a chi-square test was performed.
Measurements regarding fetuses were based on a litter as a sample unit.

Indices:

The following rates (indices) were calculated:
Implantation rate = (No. of implantations / corpora lutea counts) x 100
Embryonic/fetal mortality rate = (No. of embryonic/fetal deaths / No. of implantations) x 100
Rate of implantation sites = (No. of implantation sites / No. of implantations) x 100
Rate of placental remnants = (No. of placental remnants / No. of implantations) x 100
Rate for early (late) fetus resorption = (No. of fetuses resorbed / No. of implantations) x 100
Pre-implantation mortality = [(corpora lutea counts - No. of implantations) / corpora lutea counts] x 100
Post-implantation mortality = (No. of embryonic/fetal deaths / No. of implantations) x 100

Historical control data:

No data

Details on maternal toxic effects:

Maternal toxic effects:no effects. Remark: No systemic maternal toxicity, but mild skin reactions were observed at high dose (see below).

Details on maternal toxic effects:
At 5% w/v, very mild skin reactions (max Draize score 0.12) were observed in dams from day 10-12 of pregnancy. At 15% w/v, mild skin reactions (max Draize score 1.20) were observed in dams from day 8-14 of pregnancy.

Dose descriptor:

NOAEC

Effect level:

>= 15 other: % w/v

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

>= 101.3 mg/kg bw/day

Based on:

act. ingr.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEC

Effect level:

>= 15 other: % w/v

Based on:

act. ingr.

Basis for effect level:

other: developmental toxicity

Key result

Dose descriptor:

NOAEL

Effect level:

>= 101.3 mg/kg bw/day

Based on:

act. ingr.

Basis for effect level:

other: developmental toxicity

Dose descriptor:

NOAEC

Effect level:

5 other: % w/v

Based on:

act. ingr.

Basis for effect level:

other: see 'Remarks'

Key result

Dose descriptor:

NOAEL

Effect level:

33.2 mg/kg bw/day

Based on:

act. ingr.

Basis for effect level:

other: see 'Remarks'

Abnormalities:

no effects observed

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Dose descriptor:

NOAEL

Effect level:

>= 101.3 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Basis for effect level:

other: no embryotoxic/teratogenic effects observed.

Abnormalities:

not specified

Developmental effects observed:

not specified

Maternal toxicity.

No deaths were observed in any of the study groups. No changes were observed in general toxicity symptoms during pregnancy. Skin reactions caused by primary irritation of the study substance were observed at the administration site of animals in the medium (5 w/v%) and high dose (15 w/v%) groups. At 5% w/v, very mild skin reactions (max Draize score 0.12) were observed in dams from day 10-12 of pregnancy. As these observations were very mild and very occasionally, they were not seen as significant. At 15% w/v, mild skin reactions (max Draize score 1.20) were observed in dams from day 8-14 of pregnancy. Body weight, body weight gain and feed consumption of female animals during pregnancy were comparable across all study groups. Autopsy of female animals on day 20 of pregnancy showed no influence of α-SFMe.

Intra-uterine data.

There were no statistically significant differences observed between the control and α-SFMe groups for the following: number of corpora lutea; number of implantations (implantation rate); number of live fetuses; number of embryonic/fetal deaths (embryonic/fetal mortality rate) and their classification [number of implantation sites (rate of implantation sites); number of placental remnants (rate of placental remnants); number of resorbed fetuses (rates for early and late fetus resorption)]; and pre- and post-implantation mortalities.

Fetal examinations

Fetal weights were slightly increased compared to the control at mid and high dose (by 6% and 9%, respectively, for males and by 6% and 8%, respectively, for females), but the differences were not statistically significant and the change is not an adverse effect. No statistically significant differences were observed in the sex ratio of living fetuses between the control and treated rats. No statistically significant differences were observed in weight of placenta of living fetuses between the control and treated rats.

Fetal external examination

Fetal external examination was performed on 25-26 litters and 370-385 fetuses per group. Fetal external anomalies were observed in 2, 1, 3 and 4 cases of the control and low, mid and high dose group, respectively. All of them had low incidence (≤0.8%), and the report stated that the external anomalies concerned may be considered to be naturally occurring (historical control data were however not reported). There was no statistically significant difference between the control and treated rats.

Fetal visceral examination

Fetal visceral examination was performed on 25-26 litters and 169-178 fetuses per group. Fetal external anomalies were observed in 1, 1, 1 and 3 cases of the control and low, mid and high dose group, respectively. All of them had low incidence (≤1.18%), and the report stated that the visceral anomalies concerned may be considered to be naturally occurring (historical control data were however not reported). There was no statistically significant difference between the control and treated rats.

Fetal skeletal examination

Fetal skeletal examination was performed on 25-26 litters and 195-207 fetuses per group. Fetal skeletal anomalies were observed in 0, 0, 2 (1.0%) and 1 (0.5%) cases of the control and low, mid and high dose group, respectively. There was no statistically significant difference observed between the control and treated rats. Fetal skeletal alterations were observed in 41, 35, 37 and 19 cases of the control and low, mid and high dose group, respectively. Skeletal alternations were found across study groups, including isolation, division, or shrinkage of cervical, thoracic, and lumbar vertebral bodies, shrinkage of the 13th rib, cervical ribs, and lumbar ribs. There was no statistically significant difference between the control and treated rats, and there were no remarkable trends for a possible treatment related non-statistically significant effect.

Degrees of ossification examined based on the ossification of bone throughout the bodies of fetuses were lower in the control group than the low, mid and high dose groups for the trunk, vertebral column, 4^thcaudal vertebral body (ossification 53%, 73%, 75% and 81% respectively), 5^thcaudal vertebral body (ossification 14%, 14%, 22% and 33% respectively), sternum, 5^thand 6^thsterna ossification centre (ossification 33-46%, 61-62%, 63-66% and 64-74% respectively), for the right forelimb, 5^thmetacarpal bone (ossification 36%, 57%, 69% and 77% respectively), digit coffin bones (ossification 60-64%, 86-87%, 88-91% and 91-97% respectively), and for the right forelimb, coffin bones of the toe (ossification 45-46%, 66%, 71-72% and 83-86% respectively). The author of the report stated the following: “Rats, however, are born with incomplete ossification that is completed after birth; thus, the regions mentioned above are all prone to varying degrees of ossification in fetuses at the end of pregnancy. The time differences across autopsies on the day of cesarean operations may also result in observation of varying degrees of ossification. It is also possible that litters with more fetuses can yield lower body weights and delayed development for fetuses, resulting in partially delayed ossification. In the present study, fetuses in the control group tended to have slightly lower (although not statistically significant) body weights compared with those in α-SFMe groups, which indicates that there were varying degrees of ossification. It should be noted that the degrees of ossification for fetuses in the α-SFMe groups were within the normal range, based on our background data.” This is an acceptable explanation. Moreover, increased degree of ossification is not an adverse effect.

The maternal NOAEL for local effects is 5% (equivalent to 33.2 mg/kg bw/day) based on mild irritation at the application site. The maternal NOAEL for systemic effects is ≥15%, equivalent to ≥101.3 mg/kg bw/day in the absence of maternally toxic effects at the highest test dose.The developmental NOAEL is also ≥15 % w/v equivalent to ≥101.3 mg/kg bw/day, in the absence of embryofetal toxic effects at the highest test dose.

 

 

Conclusions:

Developmental toxicity, dermal treatment (treatment site not covered) from day 7-17 of gestation: The maternal NOAEL for local effects is 5% (equivalent to 33.2 mg/kg bw/day) based on mild irritation at the application site. The maternal NOAEL for systemic effects is ≥15%, equivalent to ≥101.3 mg/kg bw/day in the absence of maternally toxic effects at the highest test dose. The developmental NOAEL is also ≥15 % w/v equivalent to ≥101.3 mg/kg bw/day, in the absence of embryofetal toxic effects at the highest test dose.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

101.3 mg/kg bw/day

Additional information

In a dermal OECD 414 test guideline study rats were exposed from day 7 to 17 of presumed gestation to 0, 1.5, 5 and 15% %w/v of a mixture of C12/14/16 (1/2/7) MES. No maternal and developmental toxicity occurred, so the NOAEL >= 15% (equivalent to 101.3 mg/kg bw/d). However, dermal effects were observed as irritation at the highest dose, resulting in a NOAEL of 5% (equivalent to 33.2 mg/kg bw/d) for local effects.

Although no data is available on the components C16MES and C18MES, it was judged that the NOAEL for developmental toxicity could be based on the data of a substance containing the same main component as the reaction mass.

Justification for classification or non-classification

Based on the data, there is no classification necessary for developmental or reproduction toxicity according to CLP Regulation 1272/2008.