Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
toxicity to reproduction
Remarks:
other: OECD 421 Reproductive Screeing test
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2007-10-03 till 2008-06-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study under GLP without deviations
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Füllinsdorf / Switzerland
- Age at study initiation: (P) 11 weeks wks
- Weight at study initiation: (P) Males: 291 to 333 g; Females: 172 to 207 g
- Housing: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Kliba Nafag 3433 rat / mouse maintenance diet, ad libidum
- Water (e.g. ad libitum): Community tap-water from Füllinsdorf, ad libidum
- Acclimation period: Minimum 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
For groups 2 and 3: dose formulations were prepared every second week.
Fro group 4: on the first day of the study it was apparent that mixing a batch large enough to cover this length of treatment resulted in the incorporation of a lot of air. From day 2 of the study dose formulation was therefore mixed daily.

VEHICLE
- Concentration in vehicle: no data
- Amount of vehicle (if gavage): 10 mL/kg
- Purity: Milli-Q-water
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 14 days maximum
- Proof of pregnancy: vaginal plug and sperm in vaginal smear referred to as day 0 post coitum
- After 14 days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: no
- Any other deviations from standard protocol: none
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Dose formulations were stored in the refrigerator (2 - 8 °C) in sealed glass containers.

For groups 1, 2 and 3: One mL samples were taken in triplicate from the top, middle and bottom (9 in total) of the treated formulation during stirring.
Five 1 mL samples were taken from the vehicle control group (5 in total). Following 14-days refrigerated storage of the formulations 1 mL samples were taken from the middle of each formulation during stirring and from the vehicle control to confirm achived stability. After dosing on day 1 of each preparation, the residual formulation was stored frozen. From the formulations mixed on each subsequent occasion five 1 mL samples were taken from the middle of each formulation during stirring and from the vehicle control. These samples were used to confirm achieved concentration only.

For group 4: On day 1 of the treatment period, samples were taken from the dose preparation made for 14 days of treatment for homogeneity and concentration. On day 2 of the treatment period, homogeneity and concentration samples were taken when the dose formulation was mixed for the first daily preparation, as described above. Further, on days 7, 10, 13 and 15 of the treatment period and thereafter every two week (to coincide with preparation of dose for groups 2 and 3), samples were taken to confirm the achieved concentration. Five 1 mL samples were taken from the middle (5 in total) of the formulation on each occasion of sampling for achieved concentration.
Duration of treatment / exposure:
Males: 28 days
Females: approx 70 days (approx 10 weeks) (sacrificed on day 22 post partum)
Frequency of treatment:
Males were treated over a 15-day pre-pairing period and during the pairing period up to one day before necropsy.
Females were treated throughout the pre-pairing, pairing, gestation and lactation period up to weaning on day 21 post partum.
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:
no data
Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:
no data
Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:
no data
No. of animals per sex per dose:
12 males and 12 females per group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Selected by the Sponsor based on previous rat toxicity data.
Positive control:
None
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: dailiy /(recorded from treatment start to day of necropsy).


FOOD CONSUMPTION: yes
- Time schedule:
Males: Weekly during pre-pairing and after pairing periods
Females: Pre-pairing period days 1-8 and 8-15; gestation days 0-7, 7-14 and 14-21 post coitum, and lactation days 1-7 and 7-14 post partum
No food consumption was recorded during the pairing period.

OTHER:
Vaginal Smears: Throughout pre-pairing and pairing until the smear was sperm-positive or a copulation plug was observed.
Oestrous cyclicity (parental animals):
Yes
Sperm parameters (parental animals):
Parameters examined in [F1] male parental generations: testis weight, epididymis weight, spermatogenicity, prostata and seminal vesicles (histopathology)
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, body weight, external examination

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals were sacrificed after end of mating period
- Maternal animals: All surviving animals were sacrificed on day 22 post partum (at the end of the waening period)


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations with special attention at the organs of the reproductive system.


HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated below were prepared for microscopic examination and weighed, respectively:
- gross lesions
- testes
- epididymes
- prostate
- seminal vesicles
- ovaries
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 21 days of age.
- These animals were subjected to postmortem examinations (macroscopic) as follows:


GROSS NECROPSY
- Gross necropsy consisted of external examinations

HISTOPATHOLOGY / ORGAN WEIGTHS: no
Statistics:
Statistical methods used to analyze food consumption, body weight, macroscopical findings, organ weights and reproduction data:
• Means and standard deviations of various data were calculated and included in this report.
• The Dunnett-test (many to one t-test) based on a pooled variance estimate, applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test (many-one rank test), applied instead of the Dunnetttest when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test, applied if the variables could be dichotomized without loss of information
Reproductive indices:
fertility indices, mean precoital time, post-implantation losses
Offspring viability indices:
Mean litter size, pup sex ratios and viability indices.
Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
All animals survived until the scheduled necropsy, except one female at 1000 mg/kg, which died after treatment on day 14 of the pre-pairing period. The death was considered that due to a reflux the test item went into the lung (possibly due to a dosing error).
At 1000 mg/kg,one male pushed its head through the bedding and had salivation on days 14 and 15 of the pre-pairing period. Male no. 41 had ruffled fur starting on day 6 of the pairing period and seven days later also a hunched posture until the end of the study. Due to the isolated occurance these findings were considered to be not test item-related.
At 100 mg/kg, one male had a hairless area on its front leg during the pairing and after pairing period. At 1000 mg/kg, one female had a wound on its shoulder on day 2 of the gestation period onwards, which developed a scab and then a hairless area until the end of the study. These findings were considered to be incidental.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males: Mean body weight gain was slightly reduced during the study at 300 and 1000 mg/kg, and recovered in the after pairing period at 300 mg/kg but not at 1000 mg/kg. At 300 and 1000 mg/kg, mean body weight was statistically significantly reduced from days 13 and 15 of the pairing period, respectively, until the end of the study. No clear dose-dependency was noted.
Mean food consumption was slightly dose-dependently reduced from the second week of the pre-pairing period until the end of the treatment. At 1000 mg/kg in the after pairing period, the difference reached statistical significance.

Females: At 300 and 1000 mg/kg, mean body weight gain was slightly reduced during the lactation period. The difference reached statistical significance from days 10 - 13 at 1000 mg/kg. Mean body weights were not affected by treatment with the test item in all groups.
At 1000 mg/kg, mean food consumption was slightly but not statistically significantly reduced during the treatment period. At 300 mg/kg, mean food consumption was slightly but not statistically significantly reduced during the second week of the lactation period.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
The mean number of days from estrous to estrous was unaffected by treatment with the test item. Mean number of days was 4.6, 4.0, 4.1 and 4.0 days in order of ascending dose level. A shortened cycle was noted in one female at 300 mg/kg and one control female was acyclic.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
The assessment of the integrity of the spermatogenetic cycle did not provide any evidence of impaired spermatogenesis.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
The fertility rate was 100% in all groups. At all dose-levels, there were no treatment-related effects on estrous cycle, precoital time, mean duration of gestation, number of corpora lutea and implantations, post-implantation loss, pup survival or litter size from birth through to scheduled sacrifice on day 21 post partum.

ORGAN WEIGHTS (PARENTAL ANIMALS)
No significant deviations in mean organ weight of testes and epididymides were recorded which could be attributed to the treatment with the test item, since lower mean absolute weights of epididymides were considered to be in relation with lower body weights and no test item-related histopathological findings were present.

GROSS PATHOLOGY AND HISTOPATHOLOGY (PARENTAL ANIMALS)
No test item-related histopathologic findings were observed in the reproductive organs of either sex from the parental generation.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: Generation: P and F1 (migrated information)
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Remarks on result:
other: Generation: P and F1 (migrated information)
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
Neonatal mortality was generally low and considered to be unaffected by treatment with the test item.
The total numbers of pup loss during the first four days of life were 2, 2, 0 and 1 in order of ascending dose level, corresponding to 1.3, 1.4, 0.0 and 0.7% of living pups. The resulting viability indices were 98.8, 98.6, 100 and 99.3% in order of ascending dose levels.

CLINICAL SIGNS (OFFSPRING)
No findings that were considered to be test item-related were noted at first litter check or during lactation.
Two pups in the control group and at 100 mg/kg, respectively, had no milk in the stomach either at the first litter check or on day 4 post partum and died.

BODY WEIGHT (OFFSPRING)
Mean pup weights were unaffected by treatment with the test item. Mean pup weight development during lactation was unaffected by treatment with the test item.

SEXUAL MATURATION (OFFSPRING)
Not examinated

GROSS PATHOLOGY (OFFSPRING)
No test item-related findings were noted at macroscopic examination of the pups. For one pup at 100 mg/kg, which died spontaneously on day 2 post partum, an advanced austolysis was noted.

Remarks on result:
not determinable due to absence of adverse toxic effects
Reproductive effects observed:
not specified
Conclusions:
Based on these results a general NOAEL (No Observed Adverse Effect Level) was considered to be 1000 mg/kg body weight/day.
With respect to reproduction/developmental toxicity a NOEL (No Observed Effect Level) was considered to be 1000 mg/kg/day.
Executive summary:

The purpose of this study was to generate preliminary information concerning the effects of MILLED SLI (76) on male and female reproductive performance such as gonadal function, mating behavior, conception and parturition.

Four groups of 12 males and 12 females were treated by gavage with MILLED SLI (76) once daily. The dose levels were: 0 mg/kg bw/day (control group), 100 mg/kg bw/day, 300 mg/kg bw/day, and 1000 mg/kg bw/day. Males were treated over a 15-day pre-pairing period and during the pairing period up to one day before necropsy. Females were treated throughout the pre-pairing, pairing, gestation and lactation period up to weaning on day 21 post partum. A standard dose volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (Milli-Q-water).

All animals survived until scheduled necropsy, except one female at 1000 mg/kg, which died, possibly due to a dosing error.

In males at 300 and 1000 mg/kg, mean food consumption was slightly reduced. Additionally, mean body weights were reduced from the pairing period until the end of the study, whereas no clear dose-dependency was noted. Mean body weight gain was slightly reduced and recovered in the after pairing period at 300 mg/kg but not at 1000 mg/kg.

In females at 1000 mg/kg, mean food consumption was slightly reduced during the treatment period. At 300 mg/kg mean food consumption was only reduced during the second week of the lactation period. As a result, a slight transient reduction of mean body weight gain was noted in the lactation period at both dose levels. Mean body weights were similar in all groups at the end of the study.

The fertility rate was 100% in all groups. At all dose-levels, there were no treatment-related effects on estrous cycle, precoital time, mean duration of gestation, number of corpora lutea and implantations, post-implantation loss, pup survival or litter size from birth through to scheduled sacrifice on day 21 post partum.

No significant deviations in mean organ weight of testes and epididymides were recorded which could be attributed to the treatment with the test item, since lower mean absolute weights of epididymides were considered to be in relation with lower body weights and no test item-related histopathological findings were present.

No test item-related histopathologic findings were observed in the reproductive organs of either sex from the parental generation.

No abnormal findings were noted for pups at first litter check or during the lactation period. Sex ratios at first litter check and on day 21 post partum were unaffected by treatment with the test item. Mean pup weights on day 1 post partum and mean pup weight development during the lactation period were unaffected by treatment with the test item.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
We have good data to support read across to Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, the overlapping carbon chain distribution and the common metabolite which is expected to be the source of any adverse effects, justifies the use of this study.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Quality of whole database:
We have good data to support read across to Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, the overlapping carbon chain distribution and the common metabolite which is expected to be the source of any adverse effects of this oral study. Due to the common metabolite no adjustment is considered necessary in the assessment factors due to the use of read across, a factor of 1 will be used.
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Quality of whole database:
We have good data to support read across to Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, the common metabolite is expected to be the source of any adverse effects justifies the use of this oral study. Due to the overlapping carbon chain distribution and expected common metabolite no adjustment is considered necessary in the assessment factors due to the use of read across. As the available information indicates the oral adsorption is expected to be higher than by the dermal route this will give a worst case DNEL, therefore no additional assessment factor will be applied due to the use of read across, a factor of 1 will be used.
Additional information

There is no reproductive screening test available for Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0. There is however a study on the read across substance Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, which is Klimisch 1 and was carried out according to the OECD guideline 421 under GLP, RA Unilever Reproductive Screening Test KR070193. As it has been demonstrated in-vitro, using radio labeled Sodium [14C]lauryl isethionate (Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4), that rapid hydrolysis in the gastrointestinal tract and metabolism in the liver can be expected to occur, it is considered appropriate to use this study for read across, as with Coco fatty acids 2-sulfoethyl ester, sodium salt the main exposure in the body will be to the same metabolite sodium isethionate. This study with the read across substance showed no evidence of any adverse effect on fertility or any related parameters; the NOAEL was the highest dose, a limit dose of 1000mg/kg bodyweight/day.

Short description of key information:

There is no reproductive screening test available Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0.  There is however a  study on the read across substance Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4  The study is Klimisch 1 and was carried out according to the OECD guideline 421 under GLP, RA Unilever Reproductive Screening Test KR070193.  As it has been demonstrated in-vitro, using radio labeled Sodium [14C]lauryl isethionate (Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt (sodium CAS No 85408-62-4), that rapid hydrolysis in the gastrointestinal tract and metabolism in the liver can be expected to occur. Therefore it is considered appropriate to use this study for read across, as also for Coco fatty acids 2-sulfoethyl ester, sodium salt the main exposure in the body will be to the same metabolite sodium isethionate

Justification for selection of Effect on fertility via oral route:

There is no reproductive screening test available Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0.  There is however a  study on the read across substance Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt (sodium Lauryl Isethionate) CAS No 85408-62-4, which is Klimisch 1 and was carried out according to the OECD guideline 421 under GLP, RA Unilever Reproductive Screening Test KR070193.  Rapid hydrolysis in the gastro intestinal tract together with metabolism in the liver has been demonstrated in-vitro (see toxicokinetics section) with Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4 (sodium lauryl isethionate).  Such that systemic exposure to this substance would be extremely low, with exposure being to the metabolite sodium isethionate.  There is considerable overlap in the carbon chain distribution of Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4 with Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) but the lower proportion of C18 in the Coco fatty acids 2-sulfoethyl ester, sodium salt is expected to result in the same or even more rapid hydrolysis and metabolism in the liver.  Therefore it is considered appropriate to use this study for read across, for Coco fatty acids 2-sulfoethyl ester, sodium salt as the main exposure in the body for both substances would be to the same metabolite sodium isethionate (sodium 2-hydroxyethanesulfonate CAS No 1562-00-1).

Justification for selection of Effect on fertility via inhalation route:

We do not have an inhalation study investigating possible reproductive effects of Coco fatty acids 2-sulfoethyl ester, sodium salt but we do have a read across study using the oral route of exposure.  The REACH guidance provides a process to allow an inhalation DNEL to be calculated based on the oral NOAEL for the read across substance Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4.  There is an adjustment by a factor of 2 to allow for expected higher absorption via inhalation compared to ingestion.  Therefore it is not considered scientifically justified to perform an animal study by the inhalation route.

Justification for selection of Effect on fertility via dermal route:

We do not have a dermal study investigating possible reproductive effects of Coco fatty acids 2-sulfoethyl ester, sodium salt but we do have a read across study using the oral route of exposure.  The REACH guidance provides a process to allow a dermal DNEL to be calculated based on the oral NOAEL.  The available information on the dermal penetration of the read across substance Fatty acids, C12-18 and C18-unsaturated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4 indicates that Coco fatty acids 2-sulfoethyl ester, sodium salt would only be absorbed through the skin to a low to moderate extent, therefore it is expected that the oral NOAEL for the read across substance would be lower than for dermal exposure, so this will give a conservative value.

Effects on developmental toxicity

Description of key information

There are no developmental toxicity study available for Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0.  There are however two studies on the read across substance Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, a pre-natal developmental toxicity study and a reproductive screening study, which are Klimisch 1 and were carried out according to the OECD guidelines 414 and 421 respectively under GLP, RA Unilever Prenatal Development Study KT070195 and RA Unilever Reproductive Screening Test KR070193.  As it has been demonstrated in-vitro, using radio labeled Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt (sodium Lauryl Isethionate) CAS No 85408-62-4, that rapid hydrolysis in the gastrointestinal tract and metabolism in the liver can be expected to occur, it is considered appropriate to use this study for read across, for Coco fatty acids 2-sulfoethyl ester, sodium salt as the main exposure in the body for both substances would be to the same metabolite sodium isethionate.  

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2008-01-07 till 2008-04-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study under GLP without deviations
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd., Füllinsdorf / Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: 187 to 237 g
- Fasting period before study: no data
- Housing: Individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libidum
- Water (e.g. ad libitum): Community tap-water from Füllinsdorf, ad libidum
- Acclimation period: 7 days


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

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
For groups 2 and 3: every second week
For group 4: daily.

DIET PREPARATION:
not applicable (administration by gavage)


VEHICLE
- Concentration in vehicle: no data
- dose volume applied: 10 mL/kg
- Amount of vehicle (if gavage): 10 mL/kg
- Purity: Milli-Q-water
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For groups 1, 2 and 3:
The formulations mixed for the first two weeks of the study were sampled to confirm achieved concentration and homogeneity. One ml samples were taken in triplicate from the top, middle and bottom of the treated formulation during stirring. Five 1 mL samples were taken from the vehicle control group. After dosing on day 1 of each preparation, the residual formulation were stored frozen. From the formulations mixed on each subsequent occasion, five 1 mL samples were taken from the middle of each formulation during stirring and from the vehicle control. These samples were used to confirm achieved concentration only.
For group 4:
The formulation mixed for the first day of treatment was sampled to confirm achieved concentration and homogeneity. One ml samples were taken in triplicate from the top, middle and bottom of the formulation during stirring. Thereafter every two weeks samples were taken to confirm achieved concentration. Five 1 mL samples were taken from the middle of the formulation during stirring on each occasion.

All formulation samples (for all groups) despatched to the Principal Investigator were transferred to an amber glass vial and frozen (-20 ± 5 °C) pending analysis by Liquid Chromatography/Mass Spectrometry (LC/MS).
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: until evidence of copulation was observed
- Further matings after two unsuccessful attempts: no data
- Verification of same strain and source of both sexes: no (these male rats were in the possession of RCC)
- Proof of pregnancy: vaginal plug or sperm in vaginal smear, referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no
Duration of treatment / exposure:
from day 6 post-coitum to day 20 post-coitum
Frequency of treatment:
Daily
Duration of test:
20 days (day 21: females sacrificed and fetuses removed by Caesarean section)
Remarks:
Doses / Concentrations:
100 mg/kg bw/day
Basis:

Remarks:
Doses / Concentrations:
300 mg/kg bw/day
Basis:

Remarks:
Doses / Concentrations:
1000 mg/kg bw/day
Basis:

No. of animals per sex per dose:
22 females per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on a previous Reproduction/Developmental Toxicity Screening Test in the Han Wistar Rat, RCC Study Number B57982, using dose levels of 100, 300, and 1000 mg/kg/day, which showed no adverse effects on reproduction.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily


DETAILED CLINICAL OBSERVATIONS: No data


BODY WEIGHT: Yes
- Time schedule for examinations: daily from day 0 until day 21 post coitum


FOOD CONSUMPTION: Yes
- Time schedule: recorded at 3-day intervals: days 0-3, 3-6, 6-9, 9-12, 12-15, 15-18 and 18-21 post coitum.


POST-MORTEM EXAMINATIONS: Yes (gross macroscopic examination)
- Sacrifice on gestation day 21 post coitum
- Organs examined: all internal organs with emphasis to on the uterus, uterine contents, position of fetuses in the uterus and the number of corpora lutea

Ovaries and uterine content:
The uterine content was examined after termination: Yes
The ovaries content examined after termination: No
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
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
Statistics:
- Means and standard deviations of various data
- The Dunnett-test (many to one t-test) based on a pooled variance estimate, applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test), applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test, applied if the variables could be dichotomized without loss of information.
Indices:
No data
Historical control data:
Yes
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
General Tolerability - All females survived until the scheduled necropsy. No signs of discomfort or clinical symptoms from the treatment with the test item were observed.
Food Consumption and Body Weights - Mean food consumption, mean body weight and corrected body weight gain (corrected for the gravid uterus weight) were not affected by treatment with the test item in any dose group.
Reproduction Data - Post-implantation losses and the mean number of fetuses per dam were not affected by treatment with the test item at any dose level.
Macroscopical Findings - No macroscopical findings were noted during necropsy of the females.
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
External Examination - During the external examination of the fetuses, no test item-related abnormal findings were noted.
Sex Ratios - No test item-related effects on fetal sex ratios were noted in any dose group.
Body Weights - No test item-related effects on fetal body weights were noted.
Visceral Examination - No test item-related abnormalities were noted during the visceral examination of fetuses.
Skeletal and Cartilage Examination - No abnormalities, which were considered to be test item-related, were noted during examination of fetal skeletons and cartilages.
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
not specified
Developmental effects observed:
not specified

Maternal Toxicity:

General Tolerability

All females survived until the scheduled necropsy. No signs of discomfort or clinical symptoms from the treatment with the test item were observed.

Food Consumption and Body Weights

Mean food consumption, mean body weight and corrected body weight gain (corrected for the gravid uterus weight) were not affected by treatment with the test item in any dose group.

Reproduction Data

Post-implantation losses and the mean number of fetuses per dam were not affected by treatment with the test item at any dose level.

Macroscopical Findings

No macroscopical findings were noted during necropsy of the females.

Embryotoxic/teratogenic effects:

External Examination

During the external examination of the fetuses, no test item-related abnormal findings were noted.

Sex Ratios

No test item-related effects on fetal sex ratios were noted in any dose group.

Body Weights

No test item-related effects on fetal body weights were noted.

Visceral Examination

No test item-related abnormalities were noted during the visceral examination of fetuses.

Skeletal and Cartilage Examination

No abnormalities, which were considered to be test item-related, were noted during examination of fetal skeletons and cartilages.

Conclusions:
Based on the above mentioned results, the NOEL (No Observed Effect Level) for maternal and fetal organisms was considered to be 1000 mg/kg body weight/day.
Under the conditions described for this study, MILLED SLI (76) did not reveal teratogenic potential up to and including 1000 mg/kg body weight/day.
Executive summary:

Females were treated with MILLED SLI (76) to detect effects on the pregnant rat and development of the embryo and fetus.

Four groups of females were treated by gavage once daily (from day 6 post coitum to day 20 post coitum) at dose levels of 0 mg/kg body weight/day (control group), 100 mg/kg body weight/day, 300 mg/kg body weight/day and 1000 mg/kg body weight/day. A standard dose volume of 10 mL/kg body weight with a daily adjustment to the actual body weight was used. Control animals were dosed with the vehicle alone (Milli-Q-Water).

All females survived until the scheduled necropsy. No signs of discomfort or clinical symptoms from the treatment with the test item were observed. Mean food consumption, mean body weight and corrected body weight gain (corrected for the gravid uterus weight) were not affected by treatment with the test item in any dose group. Post-implantation losses and the mean number of fetuses per dam were not affected by treatment with the test item at any dose level. No macroscopical findings were noted during necropsy of the females.

No test item-related abnormal findings were noted during the external examination of the fetuses. No test item-related effects on fetal sex ratios were noted in any dose group. No test item-related effects on fetal body weights were noted. No test item-related abnormalities were noted during the visceral examination of fetuses. No abnormalities, which were considered to be test item-related, were noted during examination of fetal skeletons and cartilages.

Based on the above mentioned results, the NOEL (No Observed Effect Level) for maternal and fetal organisms was considered to be 1000 mg/kg body weight/day.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
We have good data to support read across to Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, the overlapping carbon chain distribution and the common metabolite which is expected to be the source of any adverse effects on development, justifies the use of this study.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Quality of whole database:
We have good data to support read across to Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, the overlapping carbon chain distribution and common metabolite which is expected to be the source of any adverse effects on developmental toxicity which justifies the use of this oral study. Due to the common metabolite no adjustment is considered necessary in the assessment factors due to the use of read across, a factor of 1 will be used.
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Quality of whole database:
We have good data to support read across to Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, the overlapping carbon chain distribution and common metabolite which is expected to be the source of any adverse effects justifies the use of this oral study. Due to the common metabolite no adjustment is considered necessary in the assessment factors due to the use of read across. Also the available information indicates the oral adsorption is expected to be higher than by the dermal route this will give a worst case DNEL; therefore an assessment factor of 1 will be applied.
Additional information

There are no reproduction or developmental toxicity study available for Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0. There are however two studies on the read across substance Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, which are Klimisch 1 and were carried out according to the OECD guideline 414 and 421 under GLP, RA Unilever Prenatal Development Study KT070195 and RA Unilever Reproductive Screening Test KR070193 As it has been demonstrated in-vitro, using radio labeled Sodium [14C]lauryl isethionate (Fatty acids, C12 -18 and C18 -unstaurated, 2 -sulfoethyl esters, sodium salt CAS No 85408 -62 -4), that rapid hydrolysis in the gastrointestinal tract and metabolism in the liver can be expected to occur, it is considered appropriate to use this study for read across, for Coco fatty acids 2-sulfoethyl ester, sodium salt which would be metabolized in the same way such that the main exposure in the body for both substances would be to the same metabolite sodium isethionate. 

These studies with the read across substance showed no evidence of any adverse effect on fertility or reproductive performance in the parental animals or in the pre-natal development and post natal development of rat embryos up to day 4 post partum, in particular no effects on fertility or other reproductive parameters in the parental rats or embryotoxic or teratogenic or other adverse effects on development in the offspring, the NOAEL in both studies for both reproductive and developmental effect were the highest doses used, the limit dose of 1000mg/kg bodyweight/day.

Justification for selection of Effect on developmental toxicity: via oral route:

There is no prenatal developmental or reproduction toxicity studies available for Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0.  There are however  studies on the read across substance Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, which are Klimisch 1 and was carried out according to the OECD guideline 414 under GLP, RA Unilever Prenatal Development Study KT070195 and the OECD guideline 421 under GLP, RA Unilever Reproductive Screening Test KR070193 .  As it has been demonstrated in-vitro, using radio labeled Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt (sodium Lauryl Isethionate) CAS No 85408-62-4, that rapid hydrolysis in the gastrointestinal tract and metabolism in the liver can be expected to occur, it is considered appropriate to use this study for read across, as also for Coco fatty acids 2-sulfoethyl ester, sodium salt the main exposure in the body will be to the same metabolite sodium isethionate.  .

Justification for selection of Effect on developmental toxicity: via inhalation route:

We do not have an inhalation study investigating possible developmental toxicity  and other reproductive effects of Coco fatty acids 2-sulfoethyl ester, sodium salt but we do have a read across studies for Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt (sodium Lauryl Isethionate) CAS No 85408-62-4 using the oral route of exposure.  The REACH guidance provides a process to allow an inhalation DNEL to be calculated based on the oral NOAEL.  There is an adjustment by a factor of 2 to allow for expected higher absorption via inhalation compared to ingestion.  Therefore it is not considered scientifically justified to perform an animal study by the inhalation route.

Justification for selection of Effect on developmental toxicity: via dermal route:

We do not have a dermal study investigating possible developmental toxicity effects of Coco fatty acids 2-sulfoethyl ester, sodium salt but we do have a read across study on Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, using the oral route of exposure.  The REACH guidance provides a process to allow a dermal DNEL to be calculated based on the oral NOAEL.  There is information on the dermal penetration of the read across substance Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4 This available information indicates that Coco fatty acids 2-sulfoethyl ester, sodium salt would only be absorbed through the skin to a low to moderate extent, therefore it is expected that the oral NOAEL would be lower than for dermal exposure, so this will give a conservative value.

Justification for classification or non-classification

There are no reproductive or developmental toxicity study available for Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0. There are however two studies on the read across substance Fatty acids, C12-18 and C18-unstaurated, 2-sulfoethyl esters, sodium salt CAS No 85408-62-4, which are Klimisch 1 and were carried out according to the OECD guideline 414 and 421 under GLP, RA Unilever Prenatal Development Study KT070195 and RA Unilever Reproductive Screening Test KR070193. As it has been demonstrated in-vitro, using radio labeled Sodium [14C]lauryl isethionate (Fatty acids, C12 -18 and C18 -unstaurated, 2 -sulfoethyl esters, sodium salt CAS No 85408 -62 -4) that rapid hydrolysis in the gastrointestinal tract and metabolism in the liver can be expected to occur. Therefore it is considered appropriate to use this study for read across, for Coco fatty acids 2-sulfoethyl ester, sodium salt as the main exposure in the body for both substances would be to the same metabolite sodium isethionate. 

In both these studies there was no evidence of any adverse effects on reproduction in the parental animals or on the prenatal and post natal development of the offspring at the 1000mg/kg bodyweight/day limit dose used. Therefore there is no requirement to classify Coco fatty acids 2-sulfoethyl ester, sodium salt (sodium cocoyl isethionate) CAS No 61789-32-0 for effects on fertility or developmental toxicity.