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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 April 2008 - 22 june 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
except during study days -6 to 9, and except for the absence of chemical analyses of dosage forms
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): LCE07104 = Alkylpolyglucoside
- Substance type: UVCB
- Physical state: white powder
- Purity: not indicated
- Impurities (identity and concentrations): not indicated
- Percentage of components: not indicated
- Purity test date: not indicated
- Lot/batch No.: T71225
- Expiration date of the lot/batch: 21 March 2009
- Stability under test conditions: not indicated
- Storage condition of test material: at room temperature, protected from sunlight

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeder
- Age at first treatment: approx. 10 weeks
- Weight at first treatment (mean): M=433 g, F=276 g
- Housing: individually, except during pairing
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Relative humidity: 50 ± 20%
- Light/dark cycle: 12h/12h
- Ventilation: 12 cycles/hour of filtered, non-recycled air.

IN-LIFE DATES: beginning: 29 April 2008 / end: up to 21 June 2008

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
test item was ground, heated to 80°C, mixed with vehicle heated to 80°C, forming a suspension.
The test item dosage forms were prepared daily

VEHICLE
- Justification for use and choice of vehicle (if other than water): lipophilicity of the substance
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg/day
- Lot/batch no. (if required): 057K6093, A0247283 and 1223873
- Purity: not indicated
Details on mating procedure:
- M/F ratio per cage: 1
- Length of cohabitation: until mating occurred
- Proof of pregnancy: vaginal plug, or sperm in vaginal smear - referred to as day 0 post-coitum
- In case of unsuccessful pairing: not detailed (pairing always succeeded)
- After successful mating each pregnant female was caged individually
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
Absence of a practical method of analysis
Duration of treatment / exposure:
from 2 weeks before mating until the end of mating (males: total of 39 days) or day 5 pp (females: total of 44-55 days)
Frequency of treatment:
once daily
Details on study schedule:
- no F1 parents (only one generation mated).
- Age at mating of the mated animals in the study: 13-16 weeks
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 300, 1000 mg/kg/day (m/f)
Basis:
other: nominal per gavage
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: no serious effects at up to 1000 mg/kg/day in a 14-day range-finding study, see 7.5.1
- Rationale for animal assignment: computerized stratification procedure (average body weight of each group is similar)
Positive control:
none

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule: day 1 and then weekly; and in F also days 0, 7, 14 and 20 post-coitum and days 1 and 5 post-partum

FOOD CONSUMPTION for each animal: Yes
- Time schedule: weekly (last 7-day consumption period)

FOOD EFFICIENCY: No

WATER CONSUMPTION: No
Oestrous cyclicity (parental animals):
Yes : from a fresh vaginal lavage, each morning during the mating period, until the females were mated
Sperm parameters (parental animals):
No seminology examinations.
Testis and epididymis : weight (all males), microscopic exmaination (some rats: see table 1).
Litter observations:
STANDARDISATION OF LITTERS: No

PARAMETERS EXAMINED:
The following parameters were examined in offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight on days 1 and 5, clinical signs
Postmortem examinations (parental animals):
SACRIFICE
- All male survivors: after the end of the mating period
- All female survivors: on day 6 post-partum

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

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- Sacrifice on non-selected breeders/progeny: not applicable (only 1 generation of parents and of offspring)
- All pups sacrificed on day 5 post-partum

GROSS NECROPSY: Yes, on pups sacrificed and found dead
- external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGTHS: Not performed
Reproductive indices:
Pre-implantation loss = 100 * (Number of corpora lutea - Number of implantation sites) / Number of corpora lutea
Post-implantation loss = 100 * (Number of implantation sites - Number of live fetuses) / Number of implantation sites
Mating index = 100 * Number of mated animals / Number of paired animals
Fertility index = 100 * Number of pregnant female partners / Number of mated pairs
Gestation index = 100 * Number of females with live born pups / Number of pregnant females
Live birth index = 100 * Number of live born pups / Number of delivered pups
Offspring viability indices:
Viability index = 100 * Number of surviving pups on day 5 post-partum / Number of live born pups

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Hypersalivation was observed in 11/20, 11/20, 16/20 and 13/20 animals treated at 0, 100, 300 or 1000 mg/kg/day, respectively (females counted during the pre-mating phase). Hypersalivation was observed mostly in or from the second week of dosing in the controls and low-dose group but was more often observed in week 1 in animals treated at 1000 mg/kg/day. During gestation slightly more females showed hypersalivation but it was observed in less females during lactation.
Reflux at dosing was observed in 0/20, 5/20, 3/20 and 7/20 animals treated at 0, 100, 300 or 1000 mg/kg/day, respectively. It was observed on one to five occasions in each affected animal and tended to occur towards the end of the dosing period.
Hypersalivation and reflux at dosing are considered to be related to treatment either with the vehicle and/or the test item but are non-adverse.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
The male groups treated at 100 or 1000 mg/kg/day gained minimally less weight than the controls during treatment period as a whole (-11% and -8%, respectively), however the scale of the difference and the absence of a dose-relationship renders the effect not toxicologically significant.
No effects were observed on body weight evolution in males treated at 300 mg/kg/day.
There were no effects on mean female body weight or body weight gains during the pre-mating, gestation or lactation phase; all values were comparable with the controls.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Mean male food consumption at 1000 mg/kg/day was slightly lower than that of the controls
during the first week of treatment. This was in part due to male R21433 which consumed
markedly less than the other males in the group. During the second week of dosing the mean food
consumption was identical to the controls so it was considered that the slightly decrease in week 1
was not related to treatment with the test item. There were no effects on mean male food
consumption at 100 or 300 mg/kg/day.
Mean food consumption of all female groups treated with LCE07104 was apparently slightly lower than that of the controls during the second week of treatment, however one control female had a particularly high consumption which skewed the group mean. It was considered that there were no effects of treatment with the test item. There were no differences between the controls and the test item-treated groups during gestation and lactation.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Bile acid concentrations are highly variable and the differences in males and females were inconsistent.
Inorganic phosphorus concentration was unaffected in males and slightly increased in females treated at 1000 mg/kg/day.
It was considered that neither of these parameters had been affected by treatment with the test item.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed

Details on results (P0)

The only clinical signs observed were hypersalivation and reflux at dosing, which were considered to be related to treatment with the test item but were non-adverse.

Effect levels (P0)

open allclose all
Key result
Dose descriptor:
NOAEL
Remarks:
parental toxicity
Effect level:
ca. 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
other: NOAEL = highest tested dose, without relevant effects.
Key result
Dose descriptor:
NOEL
Remarks:
reproduction (mating and fertility)
Effect level:
ca. 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive performance
other: NOEL = highest tested dose, without relevant effects.

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
not examined
Dermal irritation (if dermal study):
not examined
Mortality:
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
not examined

Details on results (P1)

Not applicable only 1 generation study

Results: F1 generation

General toxicity (F1)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Isolated clinical signs were observed in a few animals per group, except the group treated at 1000 mg/kg/day whose pups showed no clinical signs, but no one sign was observed consistently.
It was considered that there were no effects of treatment with the test item.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
mortality observed, non-treatment-related
Description (incidence and severity):
Pup survival was not markedly different between the control group and the groups treated with LCE07104
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
The mean body weight gain of the pups from the group treated at 100 or 1000 mg/kg/day was
slightly lower than that of the controls from day 1 to day 5 post-partum, however individual litter
values were within the control range. It was therefore considered that the apparent effect was not
related to treatment with the test item.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Details on results (F1)

Not required (no effects)

Effect levels (F1)

Key result
Dose descriptor:
NOEL
Remarks:
offspring toxicity
Generation:
F1
Effect level:
ca. 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
gross pathology
other: NOEL = highest tested dose, without relevant effects.

Results: F2 generation

General toxicity (F2)

Clinical signs:
not examined
Dermal irritation (if dermal study):
not examined
Mortality / viability:
not examined
Body weight and weight changes:
not examined
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
not examined
Histopathological findings:
not examined
Other effects:
not examined

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not examined

Details on results (F2)

Not applicable

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Not required (no effects)

Applicant's summary and conclusion

Conclusions:
Under on the experimental conditions of this study, the No Observed Adverse Effect Level (NOAEL) for parental toxicity was considered to be 1000 mg/kg/day, the No Observed Effect Level (NOEL) for reproductive performance (mating and fertility) was considered to be 1000 mg/kg/day and the NOEL for toxic effects on progeny was 1000 mg/kg/day.
Considering REACH Annex IX 8.7.2 and 8.7.3 criteria alone, a developmental toxicity study and a two-generation study in one species should be proposed as the substance is not genotoxic or toxic to reproduction according to available data, and there are no data on plasmatic exposure after dosing. However, although no exemption can be derived from these criteria, it should be underlined that it seems irrelevant to propose and perform this test, both from scientific and regulatory standpoints:
- the substance is clearly non-toxic based on the available tests including evaluations of fertility and development (as detailed above),
- it is expected to be metabolised into endogenous compounds (as discussed under 5.1),
- the expected metabolism and excretion do not suggest any bioaccumulation potential (as discussed under 5.1),
- exposure of workers is negligible when taking into account the high particle size limiting dermal and respiratory absorption (as discussed under 5.1), and the various risk management measures which are applied by SEPPIC (summarized under part A.1),
- the risk assessment for consumers of the final cosmetic product is exempted under REACH, Directive 2003/15/EC forbids in vivo testing on cosmetic ingredients for the dossier of the final cosmetic product from March 2009, and cosmetic regulations are stated to prevail over REACH requirements.
Executive summary:

The test item, LCE07104, was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before mating, during mating, gestation and until day 5 post-partum, at dose-levels of 100, 300 or 1000 mg/kg/day.

There were no unscheduled deaths during the study and the only clinical signs observed were hypersalivation and reflux at dosing, which were considered to be related to treatment with the test item but were non-adverse. There were no treatment-related effects on body weight, body weight gain or food consumption at any dose-level. There were no differences from controls for pairing, mating and fertility parameters. Pups showed no effects of treatment on survival or body weight performance.

There were no treatment-related effects on organ weights and no treatment-related macroscopic or microscopic findings were observed.

Considering REACH Annex IX 8.7.2 and 8.7.3 criteria alone, a developmental toxicity study and a two-generation study in one species should be proposed as the substance is not genotoxic or toxic to reproduction according to available data, and there are no data on plasmatic exposure after dosing. However, although no exemption can be derived from these criteria, it should be underlined that it seems irrelevant to propose and perform this test, both from scientific and regulatory standpoints:

-     the substance is clearly non-toxic based on the available tests including evaluations of fertility and development (as detailed above),

-     it is expected to be metabolised into endogenous compounds (as discussed under 5.1),

-     the expected metabolism and excretion do not suggest any bioaccumulation potential (as discussed under 5.1),

-     exposure of workers is negligible when taking into account the high particle size limiting dermal and respiratory absorption (as discussed under 5.1), and the various risk management measures which are applied by SEPPIC (summarized under part A.1),

the risk assessment for consumers of the final cosmetic product is exempted under REACH, Directive 2003/15/EC forbids in vivo testing on cosmetic ingredients for the dossier of the final cosmetic product from March 2009, and cosmetic regulations are stated to prevail over REACH requirements.