[No public or meaningful name is available]

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 Aug - 11 Nov 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

(adopted 22 January 2001)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Japenese Ministry of Agriculture, Forestry and Fisheries Testing guidelines for Toxicology studies, 12 NohSan No 8147, (24 November, 2000)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

The Department of Health of the Government of the United Kingdom

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Alcohols, lanolin

EC Number:

232-430-1

EC Name:

Alcohols, lanolin

Cas Number:

8027-33-6

Molecular formula:

UVCB

IUPAC Name:

Alcohols, lanolin

Details on test material:

- Name of test material (as cited in study report): Lanolin Alcohol
- Physical state: yellow solid
- Analytical purity: >90%
- Batch No.: 0000664004
- Expiration date of the batch: 19 Feb 2014
- Storage condition of test material: room temperature in the dark
- Other: Label: SUPER HARTOLAN-PA-(RB) 0000664004 BATCH

Test animals

Species:

rat

Strain:

other: Sprague-Dawley Crl:CD (SD) IGS BR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent, UK
- Weight at study initiation: 214 to 299 g
- Housing: The animals were housed individually in solid-floor polypropylene cages with stainless steel lids furnished with softwood flakes (Datesand Ltd., Cheshire, UK)
- Diet: pelleted diet (Rodent 2018C Teklad Global Certified Diet, Harlan UK, Oxon, UK); ad libitum
- Water: drinking water; ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 50±20
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 18 Aug 2013 To: 05 Sept 2013

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Arachis oil BP

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test item was prepared at the appropriate concentrations as a solution in Arachis oil BP. The stability and homogeneity of the test item formulations were previously determined by Harlan Laboratories Ltd., Shardlow, UK Analytical Services (Harlan Laboratories Ltd., Project Number 41301173). Results showed the formulations to be stable for at least twenty days. Formulations were therefore prepared once and stored at approximately +4°C in the dark.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples were taken of each test item formulation and were analyzed for concentration of Lanolin Alcohols at Harlan Analytical Laboratory, Shardlow. The test item concentration in the test samples was determined by HPLC with UV detection using an external standard technique. The results indicate that the prepared formulations were within ± 2% of the nominal concentration.

Details on mating procedure:

- Impregnation procedure: purchased timed pregnant
- The day that positive evidence of mating was observed was designated Day 0 of gestation.

Duration of treatment / exposure:

Day 5 - 19 of gestation

Frequency of treatment:

daily

Duration of test:

15 days

No. of animals per sex per dose:

24 (except for the intermediate treatment group: Only 23 animals were used in the intermediate treatment group as upon delivery of the animals, one female was found to have a damaged eye. This female was unsuitable to be used on the study and was humanely killed.)

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dose levels were chosen based on previous toxicity data.
- Other: The oral route was selected as the most appropriate route of exposure, based on the physical properties of the test item, and the results of the study are delieved to be of value in predicting the likely toxicity of the test item to man.

Examinations

Maternal examinations:

CLINICAL OBSERVATIONS: Yes
- Time schedule: Following arrival, all animals were examined for overt signs of toxicity, ill-health or behavioral changes once daily during the gestation period. Additionally, during the dosing period, observations were recorded immediately before and soon after dosing and one hour post dosing.

BODY WEIGHT: Yes
- Time schedule for examinations: on Day 3 of gestation (before the start of treatment) and on Days 5, 6, 7, 8, 11, 14 and 17 of gestation. Body weights were also recorded for animals at terminal kill (Day 20).

FOOD CONSUMPTION: Yes
- Time schedule for examinations: for each individual animal at Day 3, 5, 8, 11, 14, 17 and 20 of gestation.

WATER CONSUMPTION: Yes
- Time schedule for examinations: daily by visual inspection of the water bottles for any overt changes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day 20
- All animals were subjected to a full external and internal examination and any macroscopic abnormalities were recorded.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number, position and type of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Placental weight: Yes

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: all per litter
- Skeletal examinations: Yes: half per litter
- Visceral examinations: Yes: half per litter

Statistics:

Female body weight change, food consumption and gravid uterus weight: Bartlett’s test for homogeneity of variance and one way analysis of variance, followed by Dunnett’s multiple comparison test or, if unequal variances were observed, on alternative multiple comparison test.
All caesarean necropsy parameters and fetal parameters: Kruskal-Wallis non-parametric analysis of variance; and a subsequent pairwise analysis of control values against treated values using the Mann-Whitney ‘U’ test, where significance was seen.
Fetal evaluation parameters, including skeletal or visceral findings: Kruskal-Wallis nonparametric analysis of variance and Mann-Whitney ‘U’ test.
Probability values (p) are presented as follows:
p<0.001 ***
p<0.01 **
p<0.05 *
p≥0.05 (not significant)

Indices:

Pre- and post-implantation losses

Historical control data:

Historical control data from several former embryofetal development toxicity studies were provided, giving information on skeletal findings, visceral examination, and reproductive data.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: Observed effects on body weight, food consumption, clinical signs, pre-implantation loss and number of corpora lutea. However, these effects were considered not to represent an adverse effect or to be incidental and unrelated to treatment.

Details on maternal toxic effects:
Mortality: There were no unscheduled deaths (table 3).

Clinical Observations: Females treated with 1000 and 300 mg/kg bw/day showed isolated incidences of increased salivation between Days 6 and 13 (table 2). No such effects were detected in females treated with 100 mg/kg bw/day. Observations of this nature are commonly observed following the oral administration of an unpalatable test item formulation and in isolation are considered not to be of toxicological significance. One female treated with 1000 mg/kg bw/day also showed fur loss between days 16 and 20 (table 2). Observations of this nature are commonly observed in isolation are not considered to be related to test item toxicity. Therefore, no treatment-related effects were found.

Body Weight: No adverse effects on body weight development were detected. Statistical analysis of the data did not reveal any significant intergroup differences (table 3).

Food Consumption: Females treated with 1000 mg/kg bw/day showed a statistically significant increase in food consumption between Days 14 and 17 (table 4). The increase in food consumption is not considered to represent an adverse effect of treatment. Furthermore, intergroup differences on body weight were considered of no toxicological significance.

Water Consumption: Daily visual inspection of water bottles did not reveal any overt intergroup differences.

Pathological examinations: No macroscopic abnormalities were detected.

Females treated with 1000 and 300 mg/kg bw/day showed a statistically significant reduction in the number of corpora lutea (table 5). This intergroup difference was not dose-related and was considered to be incidental and unrelated to treatment due to ovulation and mating occurring prior to the administration of the test item. Females treated with 1000 mg/kg bw/day also showed a statistically significant reduction in pre-implantation loss (table 5). A reduction in this parameter is considered not to represent an adverse effect of treatment therefore the intergroup difference was considered of no toxicological importance.

Effect levels (maternal animals)

1

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

2

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Litter Data and Litter Placenta and Fetal Weights:
There were no treatment-related effects on in utero offspring survival, as assessed by the mean numbers of early or late resorptions, live litter size and post-implantation losses. There was also no adverse effect in sex ratio.
For all dose groups, there were no significant treatment-related trends in the proportion of fetuses (or litters) with evidence of external, visceral or skeletal anomalies (tables 6, 7 and 8). The type of external, visceral and skeletal anomalies, were those commonly observed for this type of study. There were no findings that were considered to represent any known malformations. Statistical analysis of the data did not reveal any significant differences.
Therefore, no treatment-related effects were detected in the uterine parameters examined, in fetal viability or in growth and development.

Effect levels (fetuses)

Key result

Remarks on result:

not determinable because of methodological limitations

Fetal abnormalities

Key result

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Table 1: Summary of female performance

	Control group	100 mg/kg bw/day	300 mg/kg bw/day	1000 mg/kg bw/day
Initial group size	24	24	23	24
Pregnant animals	22	24	20	24

 

Table 2: Summary incidence of daily clinical observations

Dose level [mg/kg bw/day]	Number of animals	Clinical observations	Number of animals showing effect (day of observation)
0	24	no abnormalities detected	-
100	24	no abnormalities detected	-
300	23	Increased salivation	3 (6, 9)
1000	24	Increased salivation	15 (6-13)
		Generalized fur loss	1 (16-20)

 

Table 3: Group mean values on pregnant females

Parameter	Control group	100 mg/kg bw/day	300 mg/kg bw/day	1000 mg/kg bw/day
Number of dams examined	22	24	20	24
Mortality of dams [%]	0.0	0.0	0.0	0.0
Body weight gain [g] Day 5–20 of gestation	121.3±15.5	125.6±15.6	124.4±16.4	126.8±14.7
Gravid uterus weight [g]	82.3±9.7#	80.3±14.6	80.3±9.3	81.2±11.2

#: n=21 for gravid uterus weight. Gravid uterus weight not recorded for one female in error.

 

 

Table 4: Group mean food consumption values

Dose level [mg/kg bw/day]	Number of dams examined	Food consumption (g/rat/day) between days of gestation
		3-5	5-8	8-11	11-14	14-17	17-20
0	22	25.6±2.0	21.0±2.2	23.0±2.6	24.7±1.9	25.7±2.2	25.2±2.1
100	24	25.0±2.2	21.8±3.4	23.3±2.6	25.2±3.4	26.1±2.6	27.2±5.0
300	20	25.5±2.5	21.5±2.2	23.0±3.3	25.2±2.8	26.9±3.6	26.5±3.9
1000	24	25.4±3.1	22.0±2.0	24.7±2.3	26.2±2.0	27.9±2.6**	27.4±3.0

** Significantly different from control: p** < 0.01

 

Table 5: Group mean litter data values

Parameter	Control group	100 mg/kg bw/day	300 mg/kg bw/day	1000 mg/kg bw/day
Number of corpora lutea	14.9±1.5	14.4±1.8	13.0±1.5***	13.5±2.0**
Number of implants	13.5±1.7	12.8±2.6	12.5±1.2	13.1±1.8
Total number of live implants	12.8±2.4	12.6±2.7	12.2±1.4	12.5±2.0
Total number of embryonic/fetal deaths	1.0±1.6	0.3±0.5	0.3±0.4	0.6±1.6
Pre-implantation loss [%]	8.8±7.9	12.0±13.7	4.0±4.9	2.8±5.5**
Post-implantation loss [%]	5.8±12.0	1.0±2.7	2.1±3.7	4.0±10.4
Total number of litters	22	24	20	24
Mean fetal weight[g]	4.073±0.156	4.157±0.196	4.217±0.156	4.166±0.262
Mean placental weight[g]	0.559±0.049	0.553±0.067	0.577±0.037	0.582±0.056

** Significantly different from control: p** < 0.01

*** Significantly different from control: p*** < 0.001

Table 6: Summary incidence of fetal external findings

Parameter	0 (Control)	100 mg/kg bw/day	300 mg/kg bw/day	1000 mg/kg bw/day
Number of foetuses examined	281	303	244	301
Small fetus	0	1	1	1
Omphalocele	1	0	0	0
Pale	0	0	1	1
Total number of foetuses with fetal external findings (% of foetuses with fetal external findings)	1 (0.4)	1 (0.4)	2 (0.8)	2 (0.7)

 

Table 7: Summary incidence of fetal visceral findings

Parameter	0 (Control)	100 mg/kg bw/day	300 mg/kg bw/day	1000 mg/kg bw/day
Number of foetuses examined	146	157	128	157
Total number of foetuses with fetal visceral findings (% of foetuses with fetal external findings)	51 (33.5)	53 (34.3)	34 (26.5)	70 (45.1)

 

Table 8: Summary incidence of fetal skeletal findings

Parameter	0 (Control)	100 mg/kg bw/day	300 mg/kg bw/day	1000 mg/kg bw/day
Number of foetuses examined	135	146	117	144
Total number of foetuses with fetal skeletal findings (% of foetuses with fetal external findings)	117 (88.6)	123 (84.7)	96 (81.5)	114 (78.8)

 

NOTE: a fetus may appear in more than one category

 

Applicant's summary and conclusion

Conclusions:

The NOAEL is considered to be >= 1000 mg/kg bw/day for maternal toxicity and developmental toxicity.

Executive summary:

In a test according to OECD 414, Lanolin alcoholswas administered by gavage to three dose groups each of twenty-four (twenty-three for the 300 mg/kg bw/day dose group) time mated rats, between Days 5 and 19 of gestation at dose levels of 100, 300 and 1000 mg/kg bw/day (Croda, 2014). A further group of twenty-four time mated females was exposed to the vehicle only (Arachis oil) to serve as a control.

Clinical signs, body weight change, food and water consumptions were monitored during the study. All females were terminated on Day 20 of gestation and subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weights, fetal weight, sex and external and internal macroscopic appearance were recorded. Half of each litter were examined for detailed skeletal development and the remaining half were subjected to detailed visceral examination.

There were no unscheduled deaths. Females treated with 1000 and 300 mg/kg bw/day showed isolated incidences of increased salivation between Days 6 and 13. No such effects were detected in females treated with 100 mg/kg bw/day. Observations of this nature are commonly observed following the oral administration of an unpalatable test item formulation and in isolation are considered not to be of toxicological significance. One female treated with 1000 mg/kg bw/day also showed fur loss between days 16 and 20. Observations of this nature are commonly observed and in isolation is not considered to be related to test item toxicity. No adverse effect on body weight development was detected and statistical analysis of the data did not reveal any significant intergroup differences. At necropsy, no macroscopic abnormalites were found.

There was no treatment related effects on in utero offspring survival, as assessed by the mean numbers of early or late resorptions, live litter size and post-implantation losses. There were also no adverse effects on pre-implantation losses or in sex ratio. Females treated with 1000 and 300 mg/kg bw/day showed a statistically significant reduction in the number of corpora lutea. This intergroup difference was not dose related and was considered to be incidental and unrelated to treatment due to ovulation and mating occurring prior to the administration of the test item. Females treated with 1000 mg/kg bw/day also showed a statistically significant reduction in pre-implantation loss. A reduction in this parameter is considered not to represent an adverse effect of treatment therefore the intergroup difference was considered of no toxicological importance.

For all dose groups, there were no significant treatment-related trends in the proportion of fetuses (or litters) with evidence of external, visceral or skeletal anomalies. The type of external, visceral and skeletal anomalies, were those commonly observed for this type of study. There were no findings that were considered to represent any known malformations. Statistical analysis of the data did not reveal any significant differences.

In conclusion, the oral administration of Lanolin Alcohols to rats by gavage, at dose levels of 100, 300 and 1000 mg/kg bw/day, did not result in any toxicologically significant adverse effects. The NOAEL was therefore considered to be >= 1000 mg/kg bw/day for maternal toxicity and developmental toxicity.