Registration Dossier

Administrative data

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- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'is not considered to pose reproductive or development concern. However, as a conservative approach,the lowest NOAEL of 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day) from the study with the isoalkylester constituent, has been considered further for hazard/risk assessment.

Link to relevant study records

Referenceopen allclose all

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:
read-across source
Qualifier:
equivalent or similar to
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 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 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- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'.

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:
From November 01, 2012 to December 13, 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
KL2 due to RA
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:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
(22 Mar 1996)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Behoerde fuer Soziales, Familie, Gesundheit und Verbraucherschutz; Hamburg, Germany
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Research Models and Services Germany GmbH, Sulzfeld, Germany
- Age at study initiation: males: 50 days; females: 60 days
- Mean weight at study initiation: males: 248.8 to 298.7 g; females: 195.2 to 228.1 g
- Fasting period before study: no
- Housing: single housing in MAKROLON cages (type III plus)
- Diet: commercial ssniff R-Z V1324 (ssniff Spezialdiäten GmbH, Soest, Germany); ad libitum
- Water: tap water; ad libitum
(Analyses of diet and water was performed.)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
VEHICLE
- Concentration in vehicle: 50, 150 and 500 mg/mL
- Amount of vehicle (if gavage): 2 mL/kg
Details on mating procedure:
- M/F ratio per cage: 1 male and 1 female animal were placed in one cage during the dark period
- Length of cohabitation: The female was placed with the same male until pregnancy had occurred or 2 weeks had elapsed.
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After 2 weeks of unsuccessful pairing replacement of first male by another male with proven fertility.
- After successful mating each pregnant female was caged singly.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For the analysis of the test item-vehicle mixtures samples were taken at the following time points and stored at ≤ -20°C until analysis:
Start of treatment period; immediately after preparation of the test item-vehicle mixtures; 8 and 24 hours after storage of the test item preparations at room temperature; end of treatment period; during treatment with the test item always before administration to the last animal of the dose level group
The following parameters were determined: linearity, accuracy, precision, sensitivity, specificity, stability at +2°C to +8°C or -20°C (0, 24, 72 and 168 hours)
Duration of treatment / exposure:
Males: The daily administration of the test item was started two weeks before mating and lasted until test day 35, which was one day before sacrifice.
Females: The daily administration of the test item was started two weeks before mating and continued to at least day 3 of lactation.
Maximum: 56 days of treatment
Frequency of treatment:
once daily; 7 days/week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a 14-day range-finding study (Leuschner, 2012)
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: before and after dosing
- Cage side observations checked: skin/fur, eyes, mucous membranes, respiratory and circulatory systems, somatomotor activity and behaviour patterns

MORTALITY AND CLINICAL SIGNS
- Time schedule: at least once daily (the frequency was increased when signs of toxicity were observed); deaths were recorded twice daily (animals which died or were sacrificed during the study were necropsied as soon as possible after exitus)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before the first exposure (to allow within-subject comparisons) and once a week thereafter
- Paramters: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, pilo-erection, pupil size, and unusual respiratory pattern); changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. self-mutilation, walking backwards)

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

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

For further systemic effects (water intake, haematology, clinical chemistry, neurobehaviour), see "Repeated dose toxicity: oral" (chapter 7.5.1)

OTHER
Reproduction paramters: number of pregnant females, pre-coital time, gestation length
Sperm parameters (parental animals):
Parameters examined in P male parental generation:
testis weight, epididymis weight, and qualitative sperm staging
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of all pups/litter

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

GROSS EXAMINATION OF DEAD PUPS:
Yes, for external and internal abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [males were sacrificed on day 36]
- Maternal animals: All surviving animals [females were sacrifices on day 4 post-partum or shorty thereafter]

GROSS PATHOLOGY: Yes
- Organ weights: epididymes and testicles (all males); adrenal gland, brain, heart, kidney, liver, spleen, thymus (5/sex/dose)
- Fixation: epididymis, gross lesions, mammary gland, ovary, prostate, seminal vesicle, testicle, uterus (incl. cervix and oviducts), vagina (all animals); adrenal gland, bone marrow (os femoris), brain (cerebrum, cerebellum, brain stem), heart (left and right ventricle, septum), intestine, small (duodenum, jejunum, ileum, incl. Peyer's patches, Swiss roll method), intestine, large (colon, rectum), kidney and ureter, liver, lungs (with mainstem bronchi and bronchioles), preserved by inflation with fixative and then immersion, lymph node (1 cervical, 1 mesenteric), nerve (sciatic), oesophagus, spinal cord (3 sections), spleen, stomach, thyroid (incl. parathyroids), thymus, tissue masses or tumours (incl. regional lymph nodes), tongue (incl. base), trachea (incl. larynx), urinary bladder (5/sex/dose)
HISTOPATHOLOGY: Yes, all organs that were included for fixation (5/sex of control and high dose group)
Postmortem examinations (offspring):
SACRIFICE
- The surviving F1 offspring were sacrificed at 4 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations]
Dead pups and pups sacrificed at day 4 post-partum, or shortly thereafter, were carefully examined externally for gross abnormalities.

HISTOPATHOLOGY / ORGAN WEIGTHS
not performed
Statistics:
STUDENT's t-test (p ≤ 0.01): all numerical functional tests
Multiple t-test based on DUNNETT (p ≤ 0.05 and p ≤ 0.01): body weight, food consumption, haematology, clinical chemistry, absolute and relative organ weights
For all numerical values homogeneity of variances was tested by using the BARTLETT chi-square test. If the variances were homogeneous, the DUNNETT test (p ≤ 0.01) was used to compare the experimental groups with the control group. In case of heterogeneity of variances, the STUDENT's t-test was carried out; limit of significance was p ≤ 0.01.
Reproductive indices:
For each group the gestation index was determined:
- Fertility Index female [%] = Number of pregnant rats/Number of females used x 100
- Gestation Index [%] = Number of litters with live pups/Number of pregnant rats x 100
Offspring viability indices:
For each litter and group the following indices were determined:
- Birth Index [%] = Total number of pups born (live + dead)/Number of implantation scars x 100
- Live Birth Index [%] = Number of pups born alive on day 0/1 Total number born (live + dead) x 100
- Viability Index [%] = Number of pups alive on day 4/Number of pups live on day 0/1 x 100
- Pre-implantation loss [%] = Corpora lutea - implantations/Corpora lutea x 100
- Post-implantation loss [%] = Implantations - number of pups born alive/Implantations x 100
Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
slight reduction in body weight and food consumption of the high dose females
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
slight reduction in body weight and food consumption of the high dose females
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:
no effects observed
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
increase in post implantation loss, decrease in birth index, increase in the number of stillbirths (all high dose group)
CLINICAL SIGNS AND MORTALITY
Piloerection was seen in 1 female of the high dose group at on day 2-4 of lactation.

BODY WEIGHT AND WEIGHT GAIN AND FOOD CONSUMPTION
Reduction in body weight (-9.7%) in high dose females during lactation period. Reduction in food intake (-21.7%) in high dose females during gestation/lactation.

ORGAN WEIGHTS
All effects observed (slight increase in absolute and relative liver weight in males) were still within the historical control data of the laboratory and thus not of toxicological relevance.

GROSS PATHOLOGY
No effects observed.

HISTOPATHOLOGY
No effects observed.

REPRODUCTION
The qualitative sperm staging revealed no test item-related specific spermatogenic changes in the male animals from the high dose group (1000 mg/kg b.w./day).
- Pre-coital time: No effects observed
- Gestation length: No effects observed
- Reproduction parameters of the dams: statistically significant increase in post implantation loss, non-statistically significant decrease in birth index, elevated number of stillbirths, leading to a statistically significant reduction in the live birth index.
No effects were found for the fertility index, the gestation index and the preimplantation loss.
Key result
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: adverse effects on body weight and body weight gain and food consumption at 1000 mg/kg bw/d
Remarks on result:
other: (i.e., equivalent to 233.4 mg a.i./kg bw/day)
Key result
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: no adverse effects observed
Remarks on result:
other: (i.e., equivalent to 778 mg a.i./kg bw/day)
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: an increase in post-implantation loss, a decrease in the birth index and a decrease in the live birth index at 1000 mg/kg bw/d
Remarks on result:
other: (i.e., equivalent to 233.4 mg a.i./kg bw/day)
Key result
Dose descriptor:
NOAEL
Remarks:
reproduction
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: no spermatogenic changes
Remarks on result:
other: (i.e., equivalent to 778 mg a.i./kg bw/day)
Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Description (incidence and severity):
reduced viability (high dose group)
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
reduced litter weight of the high dose female offspring
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
A test item-related decrease in the viability index was noted in the high dose group (1000 mg/kg bw/day) (71.3 % in the high dose group vs. 98.7% in the control group). The high number of dead pups in the high dose group was due to 2 dams with no surving pups on lactation day 4 (deaths partly due to cannibalization). The total litter loss in 2 of 7 dams (28.6%) was considered as test item related.

LITTER WEIGHT
A non-statistically significant reduction in mean litter weight on lactation day 1 by 17.2% was noted in the high dose group (high dose group), which is regarded to be test item-related. Total litter weight was also reduced by 24.4% in comparison to controls.

GROSS PATHOLOGY (OFFSPRING)
No effects observed.
Key result
Dose descriptor:
NOAEL
Remarks:
developmental
Generation:
F1
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: effects on litter weight and viability of the offspring at 1000 mg/kg bw/day.
Remarks on result:
other: (i.e., equivalent to 233.4 mg a.i./kg bw/day)
Key result
Reproductive effects observed:
not specified

Individual body weights (g) of females during the pre-mating and lactation period.

Control group

Day(s) relative to start

 Animal No.

1

8

15

11

196

209

212

12

217

228

243

13

210

220

213

14

217

234

244

15

3211

215

232

16

195

208

197

17

205

224

239

18

212

238

233

19

224

236

259

20

200

218

214

Mean

209

223

229

SD

9

10

19

1000 mg/kg bw/d

Day(s) relative to start

 

1

8

15

71

200

210

225

72

210

231

234

73

206

234

247

74

210

222

235

75

194

219

218

76

218

243

223

77

214

230

227

78

222

225

210

79

198

200

201

80

203

223

231

Mean

207

224

225

SD

9

12

13

Control group

Day(s) relative to littering

 

 Animal No.

1

4

11

286

309

12

323

330

13

325

311

14

331

327

15

311

322

16

282

302

17

309

327

18

312

328

19

315

331

20

300

329

Mean

309

322

SD

16

10

1000 mg/kg bw/d

Day(s) relative to littering

 

 Animal No.

1

4

71

270

281

72

339

301

73

289

309

75

289

317

77

253

247

78

280

271

79

290

296

80

293

308

Mean

288

291

SD

24

23

Relative food consumption (mg) of females between day 1 and 4 of lactation

Control group

1000 mg/kg bw/d

 Animal No.

 

11

122

71

115

12

91

72

96

13

105

73

63

14

107

75

110

15

106

77

30

16

121

78

63

17

114

79

98

18

100

80

110

19

101

 

 

20

126

 

 

Mean

109

Mean

86

SD

11

SD

30

Viability index [%], day 1 to 4 of lactation

Control group

1000 mg/kg bw/d

 Animal No.

 

 

 

11

93

71

93

12

100

72

100

13

100

73

0

14

93

74

not pregnant

15

100

75

100

16

100

76

not pregnant

17

100

77

0

18

93

78

no viable pubs

19

89

79

100

20

100

80

100

Mean

96

Mean

70

SD

4

SD

48

Summary of Live Birth Index, Pre-implantation loss, and Post-implantation loss in female animals

 

Control group

100 mg/kg bw/d

300 mg/kg bw/d

1000 mg/kg bw/d

Live Birth Index

 

 

 

 

Mean

100

100

100

86

SD

0

0

0

35

Total %1

100

100

100

91

 

 

 

 

 

Pre-implantation loss

 

 

 

 

Mean

2.4

18.3

0.6

8.1

SD

3.1

26.0

2.0

9.8

Total %2

2.5

20.3**

0.7

9.1*

 

 

 

 

 

Post-implantation loss

 

 

 

 

Mean

7.6

9.0

7.5

20.8

SD

10.5

14.8

8.8

32.9

Total %3

7.6

10.2

7.7

21.7**

*:p < 0.05 / **: p < 0.01, Chi2-test

#1: based on the total number of live born pups and the total number of pups at birth (alive and dead)

#2: based on the total number of corpora lutea and the total number of implantation sites

#3: based on the total number of implantation sites and the total number of live born pups

 


 

Conclusions:
Under the read across study conditions, NOAEL for toxicity to reproduction was determined to be 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day) and 1000 mg/kg bw/day (or 778 mg a.i./kg bw/day) for females and males, respectively. The NOAEL for development toxicity was determined to be 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day).
Executive summary:

A study was conducted to determine the repeated dose toxicity of the read across substance, 'Isodecyl oleate' (purity: 77.8%), according to OECD Guideline 422, in compliance with GLP. Three groups of ten male and ten female Sprague-Dawley rats received the read across substance daily by oral (gavage) administration before mating, through mating and, for the females, through gestation until Day 3 post-partum. The read across substance was administered as a suspension in the vehicle, corn oil, at dose-levels of 0, 100, 300 or 1000 mg/kg bw/day. Another group of ten males and ten females received the vehicle alone, under the same experimental conditions and acted as a control group. The systemic parameters for evaluating parental toxicity have been discussed in section 7.5.1 and only the reproductive and development parameters/effects have been described here. The animals were checked for macroscopic examination of the appearance and size of the gonads, adrenal glands, uterus, and accessory reproductive organs. Male animals were checked for testis weight, epididymis weight, and qualitative sperm staging. Female animals were checked for food consumption and body weight changes, number of pregnant females, pre-coital time, gestation length. Offspring (F1 generation) were observed for litter weight, number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies and dead pups were gross examined for external and internal abnormalities. F1 pups which survived were sacrificed at 4 days age, and carefully examined externally for gross abnormalities. Changes in reproduction parameters of the dam (which could be attributed maternal toxicity) and in the development of pups were seen in the high dose females and their offspring, respectively. A statistically significant increase in post-implantation loss was noted in high dose group dams along with non-statistically significant decrease in birth index and elevated number of stillbirths leading to statistically significant reduction in the live birth index. No read across substance related influence was noted for the fertility index, gestation index and the pre-implantation loss. The qualitative sperm staging revealed no read across substance related spermatogenic changes. A read across substance-related decrease in the viability index was noted in the high dose group ((71.3 % in the high dose group vs. 98.7% in the control group). The high number of dead pups in the high dose group was due to 2 dams with no surving pups on lactation Day 4 (deaths partly due to cannibalization). The total litter loss in 2 of 7 dams (28.6%) was considered as read across substance related. Further, anon-statistically significant reduction in mean litter weight on lactation Day 1 by 17.2% was noted in the high dose group, which is regarded to be test item-related. Total litter weight was also reduced by 24.4% in comparison to controls. Therefore, the NOAEL for systemic or maternal toxicity was 300 mg/kg bw/day (i.e., equivalent to 233.4 mg a.i./kg bw/day) in females, because of reduced food consumption and body weight observed in the high dose group; and at ≥ 1000 mg/kg bw/day (i.e., equivalent to 778 mg a.i./kg bw/day) in males, due to absence of any adverse effects. Further, the NOAEL for reproductive toxicity was established at 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day), due to an increase in post-implantation loss, a decrease in the live birth index at 1000 mg/kg bw/day; and the NOAEL for development toxicity was also established at 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day), due to effects on litter weight and viability of the offspring in the high dose group (Hansen, 2013). Based on the results of the read across study, NOAELs for systemic and reproductive/development toxicity can be considered for the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
233.4 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Good quality Guideline compliant studies
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), alcohol and isoalkylester. The results are presented below:

Constituent: 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: 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.

Constituent: Isoalkylester - read across study:

A study was conducted to determine the repeated dose toxicity of the read across substance, 'Isodecyl oleate' (purity: 77.8%), according to OECD Guideline 422, in compliance with GLP. Three groups of ten male and ten female Sprague-Dawley rats received the read across substance daily by oral (gavage) administration before mating, through mating and, for the females, through gestation until Day 3 post-partum. The read across substance was administered as a suspension in the vehicle, corn oil, at dose-levels of 0, 100, 300 or 1000 mg/kg bw/day. Another group of ten males and ten females received the vehicle alone, under the same experimental conditions and acted as a control group.The systemic parameters for evaluating parental toxicity have been discussed in section 7.5.1 and only the reproductive and development parameters/effects have been described here. The animals were checked for macroscopic examination of the appearance and size of the gonads, adrenal glands, uterus, and accessory reproductive organs. Male animals were checked for testis weight, epididymis weight, and qualitative sperm staging. Female animals were checked for food consumption and body weight changes, number of pregnant females, pre-coital time, gestation length. Offspring (F1 generation) were observed for litter weight, number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies and dead pups were gross examined for external and internal abnormalities. F1 pups which survived were sacrificed at 4 days age, and carefully examined externally for gross abnormalities.Changes in reproduction parameters of the dam (which could be attributed maternal toxicity) and in the development of pups were seen in the high dose females and their offspring, respectively. A statistically significant increase in post-implantation loss was noted in high dose group dams along with non-statistically significant decrease in birth index and elevated number of stillbirths leading to statistically significant reduction in the live birth index. No read across substance related influence was noted for the fertility index, gestation index and the pre-implantation loss. The qualitative sperm staging revealed no read across substance related spermatogenic changes. A read across substance-related decrease in the viability index was noted in the high dose group ((71.3 % in the high dose group vs. 98.7% in the control group). The high number of dead pups in the high dose group was due to 2 dams with no surviving pups on lactation Day 4 (deaths partly due to cannibalization). The total litter loss in 2 of 7 dams (28.6%) was considered as read across substance related. Further, anon-statistically significant reduction in mean litter weight on lactation Day 1 by 17.2% was noted in the high dose group, which is regarded to be test item-related. Total litter weight was also reduced by 24.4% in comparison to controls. Therefore, the NOAEL for systemic or maternal toxicity was 300 mg/kg bw/day (i.e., equivalent to 233.4 mg a.i./kg bw/day) in females, because of reduced food consumption and body weight observed in the high dose group; and at ≥ 1000 mg/kg bw/day (i.e., equivalent to 778 mg a.i./kg bw/day) in males, due to absence of any adverse effects. Further, the NOAEL for reproductive toxicity was established at 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day), due to an increase in post-implantation loss, a decrease in the live birth index at 1000 mg/kg bw/day; and the NOAEL for development toxicity was also established at 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day), due to effects on litter weight and viability of the offspring in the high dose group (Hansen, 2013).Based on the results of the read across study, NOAELs for systemic and reproductive/development toxicity can be considered for the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'.

Overall, based on the available weight of evidence from read across studies on the main constituents, the test substance 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate' is not considered to pose reproductive or development concern. As a conservative approach, the lowest NOAEL of 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day) from the study with the isoalkylester constituent, 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- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate' does not warrant classification for reproductive toxicity, according to EU CLP criteria (Regulation 1272/2008/EC).​​​