Reaction mass of hexadecyl dihydrogen phosphate and cetyl alcohol (mono- C16 PSE and C16-OH)

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Based on the available weight of evidence from read across studies on the main constituents, the test substance ‘mono- C16 PSE and C16-OH’ is not considered to pose reproductive or development concern up to highest tested doses ranging from 1000 -2000 mg/kg bw/day. Further, as a conservative approach,the lower NOAEL of 1000 mg/kg bw/day has been considered further for hazard/risk assessment.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

Refer to section 13 of IUCLID for details on the read-across justification. The study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar 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:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Rat Wistar aged 8 (males) - 9 (females) weeks at start of exposure period. 12M+12F/group

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

- Type of exposure: Dietary
- Duration of test/exposure: males 41-44 d, females approx. 54 d
- Treatment: Control group and treatment
- Vehicle: Diet
- Concentration in vehicle: 0, 1500, 7500 & 30,000 ppm

Details on mating procedure:

14 d premating exposure, then 1M+1F caged together. Inspection for vaginal plugs thrice daily. If mating did not occur after 14 d cohabitation the female was placed with another male for 8 d.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Actual dose received: 0, 100, 500 and 2000 mg/kg bw/day

Duration of treatment / exposure:

Males 41 - 44 d
Females up to 54 d

Frequency of treatment:

continuous in diet

Remarks:

0, 1500, 7500 & 30,000 ppm test substance in diet

Remarks:

Actual dose received: 0, 100, 500, 2000 mg/kg bw/day

No. of animals per sex per dose:

12 animals per sex per dose

Control animals:

yes

Parental animals: Observations and examinations:

Body weight, weight gain, food consumption, and food efficiency were recorded.

Oestrous cyclicity (parental animals):

Exposure was for 14 d premating covering at least 2 oestrous cycles. Ovaries were weighed and examined histopathologically at section (21 d after birth).

Sperm parameters (parental animals):

Exposure 14 d premating, no specific sperm analyses carried out, the testes & epididymis were weighed and examined histopathologically.

Postmortem examinations (parental animals):

- Organ weights P: liver, kidneys, thymus, testis, and epididymis.
- Histopathology P: liver, kidneys, adrenals, brain, heart, spleen, ovaries, thymus, testes, epididymides and any organs showing abnormality on macroscopic examination were fixed. The above tissues from all controls and top dose treated rats (except the thymus) plus abnormalities were examined.
- Macroscopic P: Full macroscopic examination.

Statistics:

Analysis of variance followed if significant differences were established by Dunnett’s T-test to assess possible intergroup differences. For pregnancy rate a Qui2-test was carried out to confirm lack of significance.

Reproductive indices:

Pregnancy rate, length of gestation, implantations, corpora lutea and resorptions were recorded.

Offspring viability indices:

Offspring (and dams) were sacrificed on post-natal Day 5 and the pups were weighed and examined for external malformations than sexed and examined for internal malformations.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Haematological findings:

no effects observed

Description (incidence and severity):

None considered of biological significance

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

None considered of biological significance

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There were no treatment related histopathological changes including no effects in the testes and ovaries.

Reproductive function: oestrous cycle:

not specified

Description (incidence and severity):

Not reported

Reproductive function: sperm measures:

not specified

Description (incidence and severity):

Not reported

Reproductive performance:

no effects observed

Description (incidence and severity):

There was no statistically significant difference in pregnancy rates although they were reduced in treated groups C 92%, 100 & 500 mg/kg bw/day 83%, 2000 mg/kg bw/day 75% these were within the normal historical control range according to the authors (actual historical control data not presented).
- Fertility index: Not reported
- Precoital interval: Not reported
- Duration of gestation: Comparable in treated and control dams (23 d in all groups).
- Gestation index: Not reported
- Changes in lactation: Not reported
- Number of implantations: No significant differences in the numbers of implantations between treated and control groups (mean 13 in control group, 14 in each treated group). There were no resorptions.
- Number of corpora lutea: No significant differences between treated and control groups (mean 14 in test and controls).
- Ovarian primordial follicle counts: Not reported

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 2 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects were observed on reproductive parameters

Critical effects observed:

no

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Haematological findings:

no effects observed

Description (incidence and severity):

None considered of biological significance

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

None considered of biological significance

Histopathological findings:

no effects observed

Description (incidence and severity):

There were no treatment related histopathological changes including no effects in the testes and ovaries.

Other effects:

no effects observed

Description (incidence and severity):

- Litter size and weights: No effect of treatment. Litter size mean Controls 13.25, low dose 13.27, mid dose 13.2, high dose 13.33. Mean litter weights at day 1 were 75, 75, 71 and 77 gm and at day 4 106, 107, 101 and 104 gm for control, low, mid and high dose respectively. No statistical significance.
- Sex and sex ratios: No treatment related effects.
- Post natal survival until Day 5: Similar in treated and control groups.

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

ca. 2 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects observed at highest tested dose

Critical effects observed:

no

Key result

Reproductive effects observed:

no

Conclusions:

Based on the results of the read across study, the test substance, mono- C16 PSE + C16-OH, NOAEL for reproductive and developmental effects is considered to be 2000 mg/kg/day.

Executive summary:

A screening study was conducted to determine the toxicity to reproduction of the read across substance, 1-dodecanol (Purity not specified) according to the method similar to the OECD Guideline 422, in compliance with GLP. The read across substance at 0, 1500, 7500 and 30000 ppm was administered as dietary feed to male Wistar rats up to 41 to 44 d, whereas to female Wistar rats up to 54 d (12 animals per sex per dose). Actual received doses were calculated to 0, 100, 500 and 2000 mg/kg bw/day. After 14 d premating exposure, one male and one female were caged together. Inspection for vaginal plugs were done thrice daily. If mating did not occur after 14 d cohabitation the female was placed with another male for 8 d. Body weight, weight gain, food consumption and food efficiency were examined. Ovaries, testes and epididymis were weighed and examined histopathologically. Pregnancy rate, length of gestation, implantations, corpora lutea and resorptions were recorded. Offspring (and dams) were sacrificed on post-natal Day 5 and the pups were weighed and examined for external malformations than sexed and examined for internal malformations. Organ weights were examined for liver, kidneys, thymus, testis, and epididymis. Liver, kidneys, adrenals, brain, heart, spleen, ovaries, thymus, testes, epididymides and any organs showing abnormality on macroscopic examination were fixed. The above tissues from all controls and top dose treated rats (except the thymus) plus abnormalities were examined. Haematological and biochemical parameters were measured. There were no treatment related effects observed on food/water consumption, body weight, organ weights, duration of gestation, biochemistry and haematological parameters. There was no statistically significant difference in pregnancy rates although they were reduced, 92% in control, 83% in 100 and 500 mg/kg bw/day, 75% in 2000 mg/kg bw/day, these were within the normal historical control range according to the study authors (actual historical control data was not presented). There were no changes attributable to exposure to the read across substance noted in gross pathology and histopathology. No significant differences in the numbers of implantations between treated and control groups (mean 13 in control group, 14 in each treated group) were noted. There were no resorptions, also no significant differences between treated and control groups (mean 14 in test and controls) were noted for number of corpora lutea. No statistical significant difference noted in litter size and weights, sex and sex ratios and post-natal survival until Day 5. No adverse effects were observed on reproductive parameters. Under the study conditions, the NOAEL for reproductive and developmental effects was established at 2000 mg/kg/day (OECD SIDS, 2006). Based on the results of the read across study, similar absence of reproductive toxicity up to 1000 mg/kg bw/day can be expected for the test substance, mono- C16 PSE and C16-OH.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Good quality Guideline compliant study

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 absence of reproductive toxicity study with the test substance, the endpoint has been assessed based on studies for substances representative of the main constituents, which can be categorised as phosphate esters (PSE) and alcohol. The results are presented below:

Constituent 1: PSE - read across study:

A screening study was conducted to determine the toxicity to reproduction of the read across substance, mono- C12 PSE, Na+ (Purity no specified) according to the OECD Guideline 422, in compliance with GLP. The test substance was administered at 0 (control group), 250, 500 or 1000 mg/kg bw to male Sprague-Dawley SPF rats for 14 d before mating, through the mating period, and up to 1 d before necropsy (42 d in total) and to female Sprague-Dawley SPF rats for 14 d before mating, through the mating period and the gestation period, up to Day 4 of lactation (42 to 45 d in total) to investigate the repeated-dose, reproductive and developmental toxicities. In the 0 and 1000 mg/kg bw groups, a 14 d recovery period was allowed after the 42 d administration period to investigate the reversibility of the toxic changes. No test substance-related effects were observed regarding clinical signs, detailed clinical findings, function tests, grip strength, amount of spontaneous movement, body weights, food consumption, urinalysis (including water intake), and haematology or blood chemistry parameters. Gross pathological examination at the end of the administration period revealed recessed areas in the forestomach at 250 mg/kg bw and above and rough mucosa or white foci at 500 mg/kg bw and above, and erosion/ulceration, mucosal thickening and submucosal edema at 250 mg/kg bw and above on histopathological examination. In the light of the absence of a forestomach in humans, observed effects on this tissue were of questionable relevance with reference to the extrapolation of the toxic properties of a test substance in humans. Further, administration of the test substance did not have any effect on the oestrous cycle, days to copulation, copulation rate, fertility rate, or conception rate. Similarly, administration of the test substance did not have any effect on the delivery rate, gestation period, number of corpora lutea, number of implantation sites, implantation rate, stillbirth rate, number of live-born pups, live-birth rate in the mother animals, or on the sex ratio of the littermates. No abnormalities were observed in the lactating behaviour during the lactation period either. These results suggest that administration of the test substance even at 1000 mg/kg bw had no effect on the reproductive function, such as that shown by the copulation rate, of the males or females, or in the fertility rate, conception rate, or on the gestation maintenance, delivery, or lactating behaviour in the mother animals. Pups showed no changes caused by the administration of the test substance regarding the observation at birth, necropsy findings on Day 4 of lactation, body weight, or viability rate, which suggested that administration of the test substance even at 1000 mg/kg bw had no effect on the development. Under the study conditions, the systemic and reproductive rat NOAELs (P0 generation) of the read across substance, were determined to be <250 and 1000 mg/kg bw/day, respectively and the rat NOAEL (F1 generation) was determined to be 1000 mg/kg bw/day (BRC, 2005).

Constituent 2: Alcohol - read across study:

A screening study was conducted to determine the toxicity to reproduction of 1-dodecanol (Purity not specified) according to the method similar to the OECD Guideline 422, in compliance with GLP. The read across substance at 0, 1500, 7500 and 30000 ppm was administered as dietary feed to male Wistar rats up to 41 to 44 d, whereas to female Wistar rats up to 54 d (12 animals per sex per dose). Actual received doses were calculated to 0, 100, 500 and 2000 mg/kg bw/day. After 14 d premating exposure, one male and one female were caged together. Inspection for vaginal plugs were done thrice daily. If mating did not occur after 14 d cohabitation the female was placed with another male for 8 d. Body weight, weight gain, food consumption and food efficiency were examined. Ovaries, testes and epididymis were weighed and examined histopathologically. Pregnancy rate, length of gestation, implantations, corpora lutea and resorptions were recorded. Offspring (and dams) were sacrificed on post-natal Day 5 and the pups were weighed and examined for external malformations than sexed and examined for internal malformations. Organ weights were examined for liver, kidneys, thymus, testis, and epididymis. Liver, kidneys, adrenals, brain, heart, spleen, ovaries, thymus, testes, epididymides and any organs showing abnormality on macroscopic examination were fixed. The above tissues from all controls and top dose treated rats (except the thymus) plus abnormalities were examined. Haematological and biochemical parameters were measured. There were no treatment related effects observed on food/water consumption, body weight, organ weights, duration of gestation, biochemistry and haematological parameters. There was no statistically significant difference in pregnancy rates although they were reduced, 92% in control, 83% in 100 and 500 mg/kg bw/day, 75% in 2000 mg/kg bw/day, these were within the normal historical control range according to the study authors (actual historical control data was not presented). There were no changes attributable to exposure to the read across substance noted in gross pathology and histopathology. No significant differences in the numbers of implantations between treated and control groups (mean 13 in control group, 14 in each treated group) were noted. There were no resorptions, also no significant differences between treated and control groups (mean 14 in test and controls) were noted for number of corpora lutea. No statistical significant difference noted in litter size and weights, sex and sex ratios and post-natal survival until Day 5. No adverse effects were observed on reproductive parameters. Under the study conditions, the NOAEL for reproductive and developmental effects was established at 2000 mg/kg/day (OECD SIDS, 2006). Based on the results of the read across study, similar absence of reproductive toxicity up to 1000 mg/kg bw/day can be expected for the test substance, mono- C16 PSE and C16-OH.

Overall, based on the available weight of evidence from read across studies on the main constituents, the test substance ‘mono- C16 PSE and C16-OH’ is not considered to pose reproductive or development concern up to highest tested doses ranging from 1000 -2000 mg/kg bw/day. Further, as a conservative approach, the lower NOAEL of 1000 mg/kg bw/day has been considered further for hazard/risk assessment.

Effects on developmental toxicity

Description of key information

See above discussion.

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 study available

Justification for classification or non-classification

Based on the available weight of evidence from read across studies on the main constituents, the test substance, ‘mono- C16 PSE and C16-OH’ does not warrant classification for reproductive toxicity, according to EU CLP criteria (Regulation 1272/2008/EC).​​​​​