Amines, C12-18-alkyl, reaction products with acrylic acid, sodium salts

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 Apr - 28 Nov 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD test guideline and GLP compliant study, performed on a test material with the same alkyl chain distribution.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 22. March 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3650, July 2000

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

The study was performed on a commercial product (aqueous solution) as the test item is manufactured and used in a liquid form.

Species:

rat

Strain:

other: HanRcc: WIST(SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories Ltd., Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: mean: males 294 - 344 g; females 176 - 221 g
- Housing: individually in Makrolon type-3 cages with wire mesh tops and sterilized standard softwood bedding
- Diet: Pelleted standard Kliba Nafag 3433 rat/mouse maintenance diet, ad libitum
- Water: community tap-water, ad libitum
- Acclimation period: 7 days

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

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

highly purified

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The dosage formulations were prepared weekly prior to administration using the test item as supplied by the Sponsor and using a factor of 3.72 taking in account the purity of 87% and the content of the active ingredient of 30.89% (CAS 3655-00-3). Sodium coco β-iminodipropionate (CAS 3655-00-3) was weighed into a glass beaker on a tared precision balance and approximately 80% of the vehicle was added (w/v). Using an appropriate homogenizer, a homogeneous suspension was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear, copulation plug observed referred to as day 0 post coitum
- After successful mating each pregnant female was caged individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Several application formulations were prepared and representative analytical samples were collected and dispatched to the analytical laboratories internally. The test item concentrations were determined by HPLC coupled to an ELSD detector and quantified with the area under the peak.

Duration of treatment / exposure:

Males: minimum of 4 weeks (2 weeks prior to mating and 2 weeks of mating)
Females: approximately 7 weeks (2 weeks prior to mating up to day 4 post partum)

Frequency of treatment:

daily, 7 days/week

Remarks:

Doses / Concentrations:
43 (group 2), 160 (group 3), 600 (group 4) mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dosage levels were selected based on a previous dose range finding toxicity study in Han Wistar Rats.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
- changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported

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

FOOD CONSUMPTION AND COMPOUND INTAKE:
- was recorded for male animals weekly during pre-pairing, for females weekly in during the pre-pairing period, gestation days 0-7, 7-14 and 14-21 post coitum, and days 1-4 post partum; food consumption was not recorded during the pairing period

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, weight gain, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for any gross anomalies

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals, after at least 28 days of dosing
- Maternal animals: All surviving animals, on day 5 post partum

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

HISTOPATHOLOGY / ORGAN WEIGHTS
For the parent animals, special attention was directed at the organs of the reproductive system. The number of implantation sites and corpora lutea was recorded for all dams with litters. The uteri of non-pregnant females were placed in a solution of ammonium sulfide to visualize possible hemorrhagic areas of implantation sites
- the following tissues from all parental males were preserved in neutral phosphate buffered 4% formaldehyde solution:
Prostate, Seminal vesicles with coagulating gland, Testes (in Bouin’s fixative), Epididymides (in Bouin’s fixative)
- the following tissues from all parental females were preserved in neutral phosphate buffered 4% formaldehyde solution:
Ovaries
- In addition, from the five males and females per group selected for organ weights, the following tissues were preserved in neutral phosphate buffered 4% formaldehyde solution:
Gross lesions, Brain (cerebrum, cerebellum, pons), Spinal chord, Small and large intestines (incl. Peyer’s patches), Stomach, Liver, Kidneys, Adrenals, Spleen, Heart, Thymus, Thyroid and Parathyroid, Trachea and lungs (preserved by inflation with fixative and then immersion), Uterus (with vagina), Urinary bladder, Lymph nodes (mandibular and mesenteric), Peripheral nerve (sciatic), Bone marrow

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring was sacrificed at day 5 post partum
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: examined macroscopically for any structural changes

Statistics:

The following statistical methods may be used to food consumption, body weight, macroscopic findings, organ weights and reproduction data:
• The Dunnett-test (many to one t-test) based on a pooled variance estimate was 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) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test was applied to the macroscopic findings.

Offspring viability indices:

birth index = number of pups born alive/number of implantations

Clinical signs:

no effects observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY
One female in group 2 died accidentally after blood sampling. This death was not considered to be test item-related.
In group 4, all animals were noted to push the head through the bedding material starting on day 9 of the pre-pairing period onwards. One female had salivation after administration of the test item on day 9 of the gestation period. These signs are considered to be signs of discomfort and not to be adverse. In groups 2 and 3, no clinical signs or observations were noted.

BODY WEIGHT AND BODY WEIGHT GAIN
- Males - pre-pairing period: In group 4, mean body weight gain was statistically significany decreased during the pre-pairing period (+6.6% versus +13.7% in the control group). Although this decrease had no statistically significant impact on the mean body weight, it was considered to be a test item related effect. During the pairing period, mean body weight and mean body weight gain were not affected by treatment with the test item. In groups 2 and 3, mean body weight and mean body weight gain were not affected by the treatment with the test item for the entire duration of the study. In group 3, mean body weight gain was statistically significant lower on day 6 of the pre-pairing period however, statistical significance occurred on a single day only and therefore it was not considered to be an adverse effect. During the pairing period, mean body weight gain was lower (+8.2% versus +12.0% in the control group) attaining statistically significance on days 9, 13 and 14. However, this was considered to be incidental as there was no-dose dependency.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean body weight gain was statistically significant decreased during the pre-pairing period (+3.5% versus +8.2% in the control group). This was considered to be a test item related effect, although it did not affect the mean body weight. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 3, mean body weight gain was lower during the pre-pairing period (+5.0% versus +8.2% in the control group) attaining statistical significance between day 6 and 8 and on days 11 and 14. This was considered an effect of the test item although mean body weight was not affected. During the gestation and lactation periods, mean body weight and mean body weight gain were not affected by treatment with the test item. In group 2, mean body weight and mean body weight gain were not affected for the entire duration of the treatment with the test item.

FOOD CONSUMPTION
- Males - pre-pairing period: In group 4, mean food consumption was statistically significant reduced between days 1-8 of the pre-pairing period (-19.2% compared to the control group) and it remained lower between days 8-14 (-10.6% compared to the control group) without attaining statistical significance. In group 3, mean food consumption was not considered to be affected by the treatment with the test item. As mean food consumption was slightly lower between days 1 and 8 of the pre-pairing period (-4.9% compared to the control group) and slightly higher between days 8 and 14 of the pre-pairing period (+2.1% compared to the control group), these variations were considered to be incidental. In group 2, no test item-related effects were noted.
- Females - pre-pairing, gestation and lactation periods: In group 4, mean food consumption was statistically significant decreased between days 1-8 of the pre-pairing period (-15.6% compared to the control group). This was considered to be a test item-related effect. During the gestation and lactation periods mean food consumption was not considered to be affected by treatment with the test item. The statistically significant higher food consumption during the lactation period (+23.0% compared to the control group) was considered to be incidental. In group 3, mean food consumption was decreased between days 1-8 of the pre-pairing period (-10.8% compared to the control group). This was considered to be a transient test item-related effect even thought it was not statistically significant. During the gestation and lactation periods, no test item-related effects were noted. In group 2, mean food consumption was not affected by treatment with the test item. During the lactation period, the statistically significant higher mean food consumption was considered to be incidental.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
All females mated within the first pairing period. The median and mean precoital times were unaffected by treatment with the test item. Mean precoital times were 4.6, 3.5, 5.3 and 4.2 days in groups 1, 2, 3 and 4, respectively. The median precoital time was 3, 3, 6 and 3 days in order of ascending dose level. The higher median precoital time noted in group 3 was considered to be incidental since there was no dose dependency.
The mean duration of gestation was unaffected by exposure to the test item. Mean duration of gestation was 21.3, 21.6, 20.7 and 21.3 days, in order of ascending dose level.
The mean number of implantations per litter and mean incidence of post-implantation loss as a percentage of total implantations were not affected by the treatment with the test item. Mean number of implantation sites was 12.9, 13.8, 13.6 and 13.4 and mean incidence of postimplantation loss was 12.1, 10.9, 7.4 and 6.6% in groups 1, 2, 3 and 4, respectively.

ORGAN WEIGHTS (PARENTAL ANIMALS)
There were no changes in organ weights for organs of the reproductive system.

GROSS PATHOLOGY (PARENTAL ANIMALS)
No gross anomalies were detected.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Histopathological evaluation of the reproductive organs did not reveal any relevant changes in high-dose animals. Special emphasis on stages of spermatogenesis and histopathology of interstitial testicular cell structure did not reveal any differences between control (group 1) and high-dose (group 4) males.

Dose descriptor:

NOEL

Effect level:

600 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects on reproductive parameters observed

Clinical signs:

not examined

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)
The birth index (number of pups born alive/number of implantations) was not affected by treatment with the test item. The birth index was 87.9, 89.1, 92.6 and 93.4% in groups 1, 2, 3 and 4, respectively. Mean litter size at first litter check was 11.3 12.3, 12.6 and 12.6 in groups 1, 2, 3 and 4, respectively. Mean postnatal loss was 0.1, 0.0, 0.2 and 0.0 in groups 1, 2, 3 and 4, respectively. Correspondingly, the viability index was 99.0, 100.0, 98.2 and 100.0%.

BODY WEIGHT (OFFSPRING)
Mean pup weights on day 1 post partum and during the lactation period (to day 4 post partum) were unaffected by treatment with the test item. On day 1 post partum mean pup weights were 5.9, 6.2, 5.9 and 5.7 g in order of ascending dose level. Mean pup weights on day 4 post partum were 8.4, 8.9, 8.2 and 8.0 g in order of ascending dose level.

SEXUAL MATURATION (OFFSPRING)
Sex ratios at first litter check and on day 4 post partum were unaffected by exposure to the test item. In group 3, the statistically significant lower number of females was considered to be incidental. The proportion of males was 45, 44, 58 and 50%, in order of ascending dose level.

GROSS PATHOLOGY (OFFSPRING)
No abnormal findings were noted at macroscopic examination of F1 pups.

Dose descriptor:

NOEL

Generation:

F1

Effect level:

600 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: within the scope of F1 generation evaluations in a screening study

Reproductive effects observed:

not specified

Conclusions:

In absence of altered parameters, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 600 mg/kg/day, the highest dose tested.

Executive summary:

The purpose of this study was to generate information concerning the effects of Sodium cocoβ- iminodipropionate (CAS 3655-00-3) on the possible health hazards likely to arise from repeated exposure over a relatively limited period of time. In addition, it provides information on possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

Sodium cocoβ-iminodipropionate (CAS 3655-00-3) was administered to male rats for at least 28 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum. The following dosage levels were administered:

Group 1: 0 mg/kg body weight/day (control group)

Group 2: 43 mg/kg body weight/day

Group 3: 160 mg/kg body weight/day

Group 4: 600 mg/kg body weight/day

Dose levels were adjusted for content of the active ingredient (30.89%) and its purity (87%), applying a correction factor of 3.72. A standard dosage 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 (highly purified water).

The NOAEL for systemic toxicity was determined to be 160 mg/kg bw. For detailed results see chapter 7.5.1 Repeated dose toxicity: oral.

No effects on reproduction or reproductive organs in parental animals were determined. The litter size and mean number of pups at first litter check and on day 4 post partum were not affected by the treatment with the test item. The sex ratio was also not affected. Mean pup weight gain was not affected by treatment with the test item. At necropsy of pups, there were no abnormal findings.

In absence of any altered parameters, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 600 mg/kg/day, the highest dose tested.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Study duration:

subacute

Species:

rat

Quality of whole database:

OECD test guideline and GLP-compliant study (Klimisch 1)

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

A study was conducted to generate information concerning the effects of sodium cocoβ- iminodipropionate (CAS # 3655-00-3) on the possible health hazards likely to arise from repeated exposure and provide information on possible effects on male and female reproductive performance such as gonadal function, mating behavior, conception, development of the conceptus and parturition, according to OECD 422 test guideline and in compliance with GLP, resulting in high reliability. Administered dose levels were 0 (control group), 43, 160 and 600 mg/kg bw/day.

 

No effects on reproduction or reproductive organs in parental animals were determined. The litter size and mean number of pups at first litter check and on day 4 post partum were not affected by the treatment with the test item. The sex ratio was also not affected. Mean pup weight gain was not affected by treatment with the test item. At necropsy of pups, there were no abnormal findings. In the absence of altered parameters, the NOEL (No Observed Effect Level) for reproduction/developmental toxicity was considered to be 600 mg/kg/day, the highest dose tested.

 

In summary results from a combined subacute toxicity / reproduction screening study did not reveal any reason of concern for offspring and for parent animals with regard to developmental toxicity or fertility up to and including the limit dose (600mg/kg bw). Since significant scientific evidence for a lack of reproduction toxic effects of the substance is drawn from these results, an additional two generation study is not expected to add any further relevant knowledge on this endpoint. No additional study is therefore proposed due to animal welfare aspects.

Short description of key information:
subacute toxicity / reproduction screening, rat, OECD 422: no adverse effects for reproduction/developmental toxicity, NOAEL (reproduction and development) >= 600 mg/kg bw/day

Justification for selection of Effect on fertility via oral route:
Only one reliable study available

Effects on developmental toxicity

Description of key information

no embryotoxic or teratogenic effects were observed in rats following dermal application of a complex formulation containing 1.5%  of an analogue substance, once every three days over the gestation period.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted similarly to OECD test guideline 414 with acceptable restrictions. The study was performed on a formulation containing an analogue substance (for justification of read-across, please refer to the corresponding assessment report in Section 13).

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

yes

Remarks:

Only one dose was tested. The animals were treated once every three days during the gestation period instead of daily.

GLP compliance:

no

Remarks:

study conducted before GLP implementation

Limit test:

yes

Species:

rat

Strain:

other: CD

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River
- Age at study initiation: no data
- Weight at study initiation: no data
- Fasting period before study: no data
- Housing: individually (the mated females)
- Diet (e.g. ad libitum): ad libitum (Ralston Purina Laboratory)
- Water (e.g. ad libitum): ad libitum (Ralston Purina Laboratory)
- Acclimation period: 2 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): controlled during the study.
- Humidity (%): controlled during the study.
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

Route of administration:

dermal

Type of inhalation exposure (if applicable):

other: not applicable

Vehicle:

water

Details on exposure:

TEST SITE
- Area of exposure: the dorso-scapular area.
- % coverage: no data
- Type of wrap if used: no data
- Time intervals for shavings or clipplings: 1 day

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no data
- Time after start of exposure: no data

TEST MATERIAL
- Concentration (if solution): 20 mL/Kg


USE OF RESTRAINERS FOR PREVENTING INGESTION: no data

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: no data
- Length of cohabitation: no data
- Further matings after two unsuccessful attempts: no data
- Verification of same strain and source of both sexes: no data
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Any other deviations from standard protocol: no data

Duration of treatment / exposure:

Days 1, 4, 7, 10, 13, 16 and 19 of gestation

Frequency of treatment:

Once every three days during the gestation period

Duration of test:

From gestation day 1 to 20 (sacrifice)

No. of animals per sex per dose:

20 pregnant rats/dose

Control animals:

yes, concurrent no treatment

other: A positive control group received acetylsalicylic acid by gavage at a dose of 250 mg/kg on days 6 through 16 of gestation.

Details on study design:

No data

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: no data


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: no data


BODY WEIGHT: Yes
- Time schedule for examinations: determined on days 1, 4, 7, 10, 13, 16 and 19 of gestation.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: no data

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Number if live and dead fetus: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one-third per litter
- Skeletal examinations: Yes: two-third per litter
- Head examinations: No data

Statistics:

The number of females exhibiting resorption sites, number of females exhibiting two or more resorptions, number of dead or resorbed fetuses, and the number of fetuses with soft-tissue or skeletal anomalies and accessory ribs was compared using chi-square test criterion or Fisher's exact probability test.
The mean number of corpora lutea, implantation sites, live fetuses, and resorption sites was compared by analysis of variance using Dunnett's multiple comparison tables to judge the significance of differences.
Statistically significant differences between groups were judged valid only when there were significant diffences between the dye treated group and each of the three untreated controls groups.

Indices:

No data

Historical control data:

Not reported

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
- No signs of toxicity were seen throughout the study, except for the changes in the color of the skin and hair at the site of dye application. No irritation or other changes in appearance were seen.
- Changes in female body weights were similar for rats in the untreated controls and the treated group. A marked reduction in maternal weight gain through gestation was observed in the rats receiving acetyl-salicylic acid (positive control) as compared with either the untreated control rats or dye-treated rats.
- Mean food consumption for all groups (treated + untreated) throughout gestation was similar except for rats in the acetylsalicylic acid group; these rats showed a moderates decrease in food consumption from days 7 to 13 of gestation. This decrease was not seen from days 13 to 20 of gestation.

Dose descriptor:

NOEL

Effect level:

2 other: mL/Kg

Based on:

other: test formulation containing 1.5% of Sodium lauriminodipropionate

Basis for effect level:

other: maternal toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
- No significant differences were observed between the treated and untreated groups in the mean number of corpora lutea, implantation sites and live fetuses and the sex ratio.
- No differences between groups were seen regarding the number of females exhibiting resorption sites or mean resorptions per pregnancy. No significant changes were observed regarding soft-tissue anomalies between the dye-treated group and the untreated control groups.
- The skeletal variations observed in this study was accessory ribs. However, this variation was more frequent in the untreated animals than the treated group.
- Increase in teratogenicity, increase in embryo-death and decrease in fetal weight were observed in the positive control group.

Dose descriptor:

NOEL

Effect level:

2 other: mL/Kg

Based on:

other: test formulation containing 1.5% of Sodium lauriminodipropionate

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Table 7.8.2/2. Summary of Teratology Study in Rats receiving p-23 (test formulation containing 1.5% of Sodium lauriminodipropionate)

Observations	Control I untreated	Control II untreated	Control III Iuntreated	Acetylsalicylic acid (250 mg/kg-day)	P-23 (2mL/Kg) (test formulation containing 1.5% of Sodium lauriminodipropionate)
Maternal parameters
Total no. females gravid	20	20	20	20	20
Mean no. corpora lutea	15.35	13.55	15.25	16.15	14.80
Mean no. implantation sites	12.4	12.10	13.90	13.25	13.50
No. females exhibiting resorption sites	13	14	12	15	12
No. females exhibiting 2 or more resorption sites	9	7	6	15	6
No. females aborting	0	0	0	0	0
Fetal parameters
Mean no.live fetuses/group	10.65	10.85	12.5	8.70	12.15
Mean live fetal weight (g)	3.38	3.58	3.62	2.89	3.96
No. dead or resorbed fetuses (%)	35 (14.11)	25 (10.33)	28 (10.07)	91 (34.34)	27 (10.00)
Mean no. resorptions/pregnancy	1.75	1.25	1.4	4.55	1.35
Sex ratio, M: F	106:107	102:115	119:131	100:75	123:120
No. fetuses with soft-tissue anomalies (%)	4 (6.35)	4 (6.06)	6 (7.79)	21 (36.84)	9 (12.16)
No. fetuses with skeletal anomalies (%)	0 (0.00)	1 (0.67)	2 (1.16)	40 (34.19)	3 (1.78)
No. fetuses with accessory ribs only (%)	75 (50.00)	56 (37.09)	72 (41.62)	32 (27.35)	66 (39.05)

 

Conclusions:

Under the test conditions of this study, no embryotoxic or teratogenic effects were observed in the CD rats following dermal application of formulation P-23 containing 1.5% of sodium lauriminodipropionate, once every three days over the gestation period.

Executive summary:

In a developmental toxicity study, one hair dye formulation containing 1.5% of sodium lauriminodipropionate was administered topically to groups of 20 CD female rats at a dose level of 2 mL/kg on days 1, 4, 7, 10, 13, 16 and 19 of gestation. Pilot study had shown that the potential skin irritancy would not permit more frequent application. Three negative control groups were tested in this study. The positive control group received acetylsalicylic acid by gavage at a dose of 250 mg/kg on days 6 through 16 of gestation.

Twenty pregnant rats from each group were sacrificed on day 20 of gestation, and Cesarean sections were performed.

No signs of toxicity were seen throughout the study, except for the changes in the color of the skin and hair at the site of dye application. No irritation or other changes in appearance were seen. Changes in female body weights were similar for rats in the untreated controls and the treated group. A marked reduction in maternal weight gain through gestation was observed in the rats receiving acetyl-salicylic acid (positive control) as compared with either the untreated control rats or dye-treated rats. Mean food consumption for all groups (treated + untreated) throughout gestation was similar except for rats in the acetylsalicylic acid group. These rats showed a moderates decrease in food consumption from days 7 to 13 of gestation. This decrease was not observed from days 13 to 20 of gestation.

No significant differences were observed between the treated and untreated groups in the mean number of corpora lutea, implantation sites and live fetuses and the sex ratio. No differences between groups were seen regarding the number of females exhibiting resorption sites or mean resorptions per pregnancy. No significant changes were observed regarding soft-tissue anomalies between the dye-treated group and the untreated control groups. The skeletal variation observed in this study was accessory ribs. However, this variation was more frequent in the untreated animals than the treated group. Increase in embryo-death and decrease in fetal weight were observed in the positive control group.

Under the test conditions of this study, no embryotoxic or teratogenic effects were observed in the CD rats following dermal application of formulation P-23 containing 1.5% of sodium lauriminodipropionate, every every three days over the gestation period.

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

Quality of whole database:

Study conducted on a complex formulation, using only one dose concentration and the dermal route of administration, considered useful together with the reproduction toxicity screening (OECD 422)

Additional information

In a developmental toxicity study, one hair dye formulation containing 1.5% of sodium lauriminodipropionate was administered topically to groups of 20 CD female rats at a dose level of 2 mL/kg on days 1, 4, 7, 10, 13, 16 and 19 of gestation. Twenty pregnant rats from each group were sacrificed on day 20 of gestation, and Cesarean sections were performed.

No significant differences were observed between the treated and untreated groups in the mean number of corpora lutea, implantation sites and live fetuses and the sex ratio. No differences between groups were seen regarding the number of females exhibiting resorption sites or mean resorptions per pregnancy. No significant changes were observed regarding soft-tissue anomalies between the dye-treated group and the untreated control groups. The skeletal variation observed in this study was accessory ribs. However, this variation was more frequent in the untreated animals than the treated group. Under the test conditions of this study, no embryotoxic or teratogenic effects were observed in the CD rats following dermal application of formulation P-23 containing 1.5% of sodium lauriminodipropionate, every every three days over the gestation period.

Despite significant limitations, this study was selected as a key study for the present endpoint. In the absence of specific toxicokinetic data, the exact dose of substance administered remains uncertain. However, this study with extensive fetal examinations can be used in a weight-of-evidence approach together with the data from the screening OECD 422 study, where no adverse findings regarding development (e.g. fetal and pup growth and survival...) were observed up to and including the limit dose (600mg/kg bw). Taken together, these data suggest that there is no concern with regards to potential developmental toxicity. The performance of a specific OECD 414 -compliant study appears to be unjustified according to Annex XI Regulation (EC) No 1907/2006 chapter 1.2 "weight of evidence .... Where sufficient weight of evidence for the presence or absence of a particular dangerous property is available:.....—further testing on vertebrate animals for that property shall be omitted, ... ", and also in line with the Title III 'Data sharing and avoidance of unnecessary testing' of Regulation (EC) No. 1907/2006.

Justification for selection of Effect on developmental toxicity: via dermal route:
Only developmental toxicity study with extensive examinations available

Justification for classification or non-classification

Based on the absence of evidence for any reproductive or developmental toxicity potential from a human or animal study, the test material does not fulfil the requirement according to GHS (Regulation (EU) 1272/2008) or DSD (67/548/EEC) to be labelled as reproductive toxicant.

 

Labelling reproductive toxicant:

GHS: no labelling

DSD: no labelling