Registration Dossier

Administrative data

Description of key information

Two repeated dose toxicity studies are available, one study following the OECD guideline n°422 with the registered substance and another one following the OECD guideline n° 408 and performed with a "close chemical structure" (acetalization product between glucose and C16/18(even numbered)-alcohol). These two studies did not show any significant adverse effect related to the compound tested.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
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:
other: compliant to GLP and testing guideline; adequate coherence between data, comments and conclusions.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
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
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
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral 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
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
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: in a previous 14-day toxicity study in the rat (see 7.5.1) no relevant adverse effects occured at the dose-levels of 100, 300 or 1000 mg/kg/day.

- Rationale for animal assignment: computerized stratification procedure (average body weight of
each group is similar)

- Satellite and post-exposure recovery period: not performed.
Positive control:
None
Observations and examinations performed and frequency:
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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on the day of sacrifice
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 5M+5F

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of sacrifice
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: 5M+5F

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule: at the end of the treatment period for M, on day 5 post-partum for F
- Dose groups that were examined: all
- Battery of functions tested: behavior / reflexes / sensory activity / grip strength / motor activity / rectal temperature
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, adults and pups of all groups
ORGAN WEIGHTS: Yes, adults of all groups
HISTOPATHOLOGY: Yes, adults in control and high-dose

all : see Table 1
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, treatment-related
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
Details on results:
CLINICAL SIGNS AND MORTALITY
There were no unscheduled deaths. Hypersalivation and reflux at dosing were observed in all groups and may be related to the vehicle and/or the test item but are non-adverse

NEUROBEHAVIOUR:
females treated at 1000 mg/kg/day had a markedly lower number of movements when compared with controls. In the absence of any relevant daily clinical signs in the females a relationship to treatment is considered unlikely.
Key result
Dose descriptor:
NOAEL
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
water consumption and compound intake
haematology
clinical biochemistry
behaviour (functional findings)
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
histopathology: neoplastic
other: see 'Remark'
Critical effects observed:
not specified

Not necessary

Conclusions:
Based 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.
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. The Functional Observation Battery assessment and hematology and blood biochemistry revealed no treatment-related effects, however females treated at 1000 mg/kg/day did make a markedly lower number of movements during the motor activity assessment when compared with the controls.

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

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
2012-11-08 to 2013-06-21
Reliability:
2 (reliable with restrictions)
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Version / remarks:
• OECD 408 “Repeated Dose 90-day Oral Toxicity Study in Rodents”, 1998. • OECD 416, “Two-Generation Reproduction Toxicity Study”, 2001.(only for clinical pathology and histopathological evaluations of male reproductive organs including sperm parameters)
Deviations:
no
Principles of method if other than guideline:
The purpose of this repeat dose toxicity study was to assess the systemic toxicity potential of the test item, Acetalization product between glucose and C16/18 (even numbered)-alcohol in Wistar rats when administered orally by gavage for 90 consecutive days and to assess the reversibility of any effects following 28-days recovery period.
This study was designed to provide information on major systemic toxic effects, target organs, the possibility of accumulation, additional parameters (clinical pathology and histopathological evaluations to evaluate effects of the test item on male reproductive organs) and an estimate of a No Observed Adverse Effect Level (NOAEL)/No Observed Effect Level (NOEL).
Detailed testicular histopathological examination was conducted in order to identify possible treatment-related effects such as retained spermatids, missing germ cell layers or types, multinucleated giant cells or sloughing of spermatogenic cells into the lumen. Intact epididymis was also examined to include the caput, corpus, and cauda, which was accomplished by evaluation of a longitudinal section. The epididymis was evaluated for possible
leukocyte infiltration, change in prevalence of cell types, aberrant cell types, and phagocytosis of sperm. As described in paragraphs 41 to 44 (OECD n°416).
GLP compliance:
yes (incl. certificate)
Remarks:
OECD Principles of Good Laboratory Practice [C (97) 186/Final]
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Wistar rats – HsdCpb: WU rats conventionally bred (In-house random bred)
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
The dose formulations were administered orally by gavage to specific group of rats once daily at approximately the same time (± 2 hours) each day for a period of 90 consecutive days. Similarly, vehicle was administered by oral gavage to rats in vehicle control and vehicle control recovery groups for 90 consecutive days. The vehicle or the dose formulations were not administered to the rats in the recovery groups for 28 Days following the 90-day treatment period.
The dose volume administered to each rat was at an equivolume of 10 mL/kg body weight throughout the study. The dose volume was adjusted based on the most recent body weight of individual rat.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The results of dose formulation (i.e. at 10, 30 and 100 mg/mL) analysis on Day 1 and during the second and third month of the treatment showed that, the mean analysed concentration of the active ingredient was within the acceptable limits (15% and RSD was less than 10%) of variation and was homogeneously mixed in the vehicle. The test item was not detected in the vehicle control group samples analysed during different intervals of the treatment period.
Duration of treatment / exposure:
The dose formulations were administered orally by gavage to specific group of rats once daily at approximately the same time (± 2 hours) each day for a period of 90 consecutive days. Similarly, vehicle was administered by oral gavage to rats in vehicle control and vehicle control recovery groups for 90 consecutive days. The vehicle or the dose formulations were not administered to the rats in the recovery groups for 28 Days following the 90-day treatment period.
The dose volume administered to each rat was at an equivolume of 10 mL/kg body weight throughout the study. The dose volume was adjusted based on the most recent body weight of individual rat.
Frequency of treatment:
Daily for 90 days
Remarks:
Doses / Concentrations:
1000 mg/kg/day
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
300 mg/kg/day
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
100 mg/kg/day
Basis:
nominal in diet
No. of animals per sex per dose:
Each main group comprised of 10 rats/sex and recovery groups 5 rats/sex as described below:


Group No. Dose (mg/kg/day) No. of Animals Dose volume (mL/kg) Terminal sacrifice
G1 Vehicle control* 10M + 10F 10 Day 91
G2 100 10M + 10F 10 Day 91
G3 300 10M + 10F 10 Day 91
G4 1000 10M + 10F 10 Day 91
G1R Vehicle control* 5M + 5F 10 Day 119
G4R 1000 5M + 5F 10 Day 119
M: Males; F: Females; * The vehicle was Olive oil
Control animals:
yes, concurrent vehicle
Details on study design:
A total of 50 males and 50 female rats were randomly allocated to four main groups and two recovery groups, each main group comprised of 10 rats/sex and recovery groups 5 rats/sex as described below:


Group No. Dose (mg/kg/day) No. of Animals Dose volume (mL/kg) Terminal sacrifice
G1 Vehicle control* 10M + 10F 10 Day 91
G2 100 10M + 10F 10 Day 91
G3 300 10M + 10F 10 Day 91
G4 1000 10M + 10F 10 Day 91
G1R Vehicle control* 5M + 5F 10 Day 119
G4R 1000 5M + 5F 10 Day 119
M: Males; F: Females; * The vehicle was Olive oil
Positive control:
no positive control
Observations and examinations performed and frequency:
Each rat was observed twice daily for mortality and morbidity during the treatment and recovery period. Routine observations for checking general clinical signs were performed once during pre-dose and two-three times during post-dose to observe the test item-related clinical sign of salivation during the treatment period and once daily during the recovery period.
Sacrifice and pathology:
All the animals of main and recovery groups were subjected to detailed necropsy and findings were recorded. The rats to be sacrificed at term were fasted overnight (water allowed), weighed, exsanguinated under isoflurane anaesthesia and subjected to detailed necropsy by a Pathologist, as per the random numbers generated for terminal sacrifice (main and recovery).
Other examinations:
Toxic effects, target organs, the possibility of accumulation, additional parameters (clinical pathology and histopathological evaluations to evaluate effects of the test item on male reproductive organs
Statistics:
The Data captured using Provantis™ for the parameters namely body weights, food consumption and organ weights were analyzed using built-in statistical tests. Derived data like net body weight change and organ weight ratios was also analyzed using above mentioned methods.
Data captured outside of ProvantisTM: The statistical analysis of the experimental data was carried out using the validated package in Excel and also using licensed copies of SYSTAT Statistical package Ver.12.0. All quantitative variables like neurological observations (neuromuscular observation and body temperature) and clinical pathology (haematology, coagulation and clinical chemistry) data was tested for normality (Shapiro-Wilk test) and homogeneity of variances (Levene’s test) within the group before performing a one-factor ANOVA modeling by treatment groups. Non-optimal (non-normal or heteroschedastic) data was transformed, before ANOVA was performed. Comparison of means between treatment groups and vehicle control group was done using Dunnett’s test when the overall treatment, ‘F’ test when found significant.
In the case of recovery groups, data was analysed using the methods stated above. Comparison of means between treatment recovery group(s) and control recovery group was performed.
All analyses and comparisons were evaluated at the 5 % (P ≤ 0.05) level. Statistically significant differences (P ≤ 0.05), indicated by the aforementioned tests was designated by the following superscripts throughout the report:
+/- : Significantly higher (+) /lower (-) than the vehicle control group
a+/a- : Significantly higher (+) /lower (-) than the vehicle control recovery group
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
• No mortalities were observed in males and females of main toxicity and recovery groups at all dose levels. • The salivation (slight to moderate) was observed in both sexes at all the test item treated groups during the course of the treatment period. This clinical sign of salivation was observed at 5-10 minutes post-dose and was persisted up to approximately 45 minutes post-dose only. The incidences of salivation were more in the high dose group rats (1000 mg/kg/day) and these were not observed consistently in same rats. The observed clinical sign of salivation was considered as transient and test item-related non-adverse effect. There were no other clinical signs observed in any of the test item treated groups.
Mortality:
mortality observed, treatment-related
Description (incidence):
• No mortalities were observed in males and females of main toxicity and recovery groups at all dose levels. • The salivation (slight to moderate) was observed in both sexes at all the test item treated groups during the course of the treatment period. This clinical sign of salivation was observed at 5-10 minutes post-dose and was persisted up to approximately 45 minutes post-dose only. The incidences of salivation were more in the high dose group rats (1000 mg/kg/day) and these were not observed consistently in same rats. The observed clinical sign of salivation was considered as transient and test item-related non-adverse effect. There were no other clinical signs observed in any of the test item treated groups.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
• Body weights, net body weight gains were unaffected by the treatment at all dose levels.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Food consumption were unaffected by the treatment at all dose levels.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
• No eye abnormalities were found in the ophthalmological examination conducted at the end of the treatment and recovery period.
Haematological findings:
no effects observed
Description (incidence and severity):
There were no test item-related changes in hematology, coagulation parameters.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
There were no test item-related changes in clinical chemistry parameters.
Urinalysis findings:
no effects observed
Description (incidence and severity):
There were no test item-related changes in urinalysis parameters.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
There were no test item-related neurological abnormalities observed at at the end of the treatment and recovery period.
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no test item-related changes in organ weights/ratios and sperm parameters.
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no test item-related gross changes among the doses tested.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no test item-related microscopic changes among the doses tested.
Details on results:
CLINICAL SIGNS AND MORTALITY
The clinical sign of salivation (slight to moderate) was observed in both sexes from Day 25 onwards in all the test item treated groups during the course of the treatment period. This clinical sign of salivation was observed at 5-10 minutes post-dose and was persisted up to approximately 45 minutes post-dose only. The incidences of salivation were more in the high dose group rats (1000 mg/kg/day). The observed clinical signs were intermittent, as they were not observed in the same rat consistently during the whole course of in-life phase. The observed clinical sign of salivation was considered as transient and test item-related non-adverse effect of the test item.

There was no mortality observed during the course of the experiment.

BODY WEIGHT AND WEIGHT GAIN
The mean body weights and net body weight gains were unaffected by the treatment at all the tested dose groups throughout the treatment period in both main and recovery group males and females.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
The food consumption was unaffected by the treatment in males at all the tested dose groups throughout the treatment period in both main and recovery group.
However, in females the following statistically significant changes were observed in the mean food consumption as compared to the vehicle control group:
- at 100 mg/kg Bwt/day, lower food consumption was observed in females during Days 57 -64,
- at 300 mg/kg Bwt/day, lower food consumption was observed in females during Days 22-29, 36-43, 57-64, 64-71 and 78-85.
- at 1000 mg/kg Bwt/day, lower food consumption was observed in females during Days 57-64, 64-71 and 78-85.
- at the high dose recovery group females, higher food consumption was observed during Days 64-71, 71- 78, 85-90, 90-97, 104-111 and 111-118 as compared to the vehicle control recovery group.
The few minor changes observed in food consumption was considered as incidental and not test item-related findings as there decreases were not associated with any changes in absolute mean body weights.

OPHTHALMOSCOPIC EXAMINATION
No eye abnormalities were found in the ophthalmological examination conducted during the acclimatization, end of the treatment and recovery period.

HAEMATOLOGY
No changes of toxicological relevance were observed in hematology parameters of both sexes. Increase in total leukocyte and lymphocyte counts noted at 1000 mg/kg/day dose recovery males were considered to be incidental, as the change was not apparent in main group rats. In addition, the magnitude of change was comparable with historical control data. (Refer Tables 1 to 4, Appendices 1 to 4 of the attached report)

No test item-related effects were noted in PT and APTT values in any of the treated groups. A decrease in APTT value noted at 1000 mg/kg/day dose recovery males was considered incidental as the change had no toxicologic relevance. (Refer Tables 1 to 4, Appendices 1 to 4 of the attached report)

CLINICAL CHEMISTRY
There were no toxicologically significant changes in biochemical parameters, in any tested groups. Increased total cholesterol (1000 mg/kg males), glucose/potassium (1000 mg/kg recovery males), and decreased GGT, ALP, total cholesterol and calcium levels (1000 mg/kg recovery males) were considered incidental, as the alterations were either comparable with in-house historical control data or lack biological significance. (Refer Tables 5 to 8, Appendices 5 to 8 of the attached report)

URINALYSIS
There were no test item-related changes in urine parameters at all the doses tested.

NEUROBEHAVIOUR
Neuromuscular Observations: There were no test item-related variations in hind limbs footsplay and fore limbs and hind limbs grip strength in both sexes.

ORGAN WEIGHTS
There were no significant changes attributed to treatment in terminal fasting body weights, organ weights and organ to body weight ratios in both the sex groups at any tested doses when compared to the vehicle control group.
The weight changes involving thymus (1000 mg/kg recovery males; ≥ 100 mg/kg females), pituitary (1000 mg/kg recovery females) and uterus (≥ 300 mg/kg females) were considered to be toxicologically not relevant, as the alterations did not show corresponding histological findings and/or were within the range of laboratory historical control data. (Refer Tables 13 to 20, Appendices 13 to 20 of the attached report)

SPERM EVALUATION (Refer Table 21, Appendices 21 to 24 of the attached report)
Sperm motility was examined for all the dose groups. There were no significant differences in percentages of progressive motile sperms and motile sperms between the groups except for the G2 group where a statistically but not biological significant decrease was observed. This observation was considered as incidental.
Sperm morphology was examined for all the dose groups. The Significantly but not statistically increased percentage of abnormal sperms noted at 1000 mg/kg/day dose was considered incidental, as the change was not dose-related and the level of abnormal sperm was near the control group values. The commonly observed abnormality across groups was headless sperms.

Cauda epididymal sperm count was done for all the dose groups. The non statistical decreased values for number of sperms per cauda epididymis and number of sperms per gram of cauda epididymis at 1000 mg/kg/day dose were due to inter-individual variations and were considered to be of no biological significance, as the alterations were not dose-related. Further, the cauda epididymal weight did not differ significantly among the treated groups.
Detergent and homogenization resistant testicular spermatic counts were performed at all the dose levels. Significantly decreased values observed at 1000 mg/kg/day involving number of spermatid per testis and number of spermatid per gram of testis were considered incidental in the absence of dose-relationship and due to an individual lower count (animal Ro3440). There were no significant differences in testicular weights between the groups. All sperm parameters evaluated remained unaffected by dosage as high as 1000 mg/kg/day.

GROSS PATHOLOGY
No changes attributed to test item were noted in treated rats of both the sexes.
Dilatation of uterus observed in a female at 100 mg/kg/day dose was microscopically confirmed and considered incidental.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no microscopic changes attributed to treatment in males and females. The microscopic findings observed in few organs in some of the animals were considered as incidental background findings and hence they were not related to test item-administration. (Refer Table 23, Appendices 29 to 32 of the attached report)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Remarks:
Acetalization product between glucose and C16/18 (even numbered)-alcohol
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified
Conclusions:
CONCLUSION
The repeated administration of test item Acetalization product between glucose and C16/18 (even numbered)-alcohol to Wistar rats through oral gavage for 90 consecutive days at the dose levels of 100, 300 and 1000 mg/kgBwt/day did not cause any toxicological effects on general health, mortality, functional observational battery, growth and food consumption. No test item-related changes observed in haematology, coagulation, clinical chemistry, urinalysis, terminal fasting body weights, organ weights/ratios and sperm parameters and gross and histopathological changes.
The slight salivation which was observed in all rats at all the doses was considered transient and test item-related non-adverse effect. Hence, under the test conditions, the evaluated “No Observed Adverse Effect Level (NOAEL)” for test item Acetalization product between glucose and C16/18 (even numbered)-alcohol is 1000 mg/kg Body weight/day under the test conditions and doses employed.
Executive summary:

The purpose of this repeat dose toxicity study was to assess the systemic toxicity potential of the test item, Acetalization product between glucose and C16/18 (even numbered)-alcohol in Wistar rats when administered orally by gavage for 90 consecutive days and to assess the reversibility of any effects following 28-days recovery period. This study was intended to provide information on major systemic toxic effects, target organs, the possibility of accumulation, additional parameters (clinical pathology and histopathological evaluations to evaluate effects of the test item on male reproductive organs) and an estimate of a No Observed Adverse Effect Level (NOAEL).

A total of 50 males and 50 female rats were randomly allocated to four main groups and two recovery groups, each main group comprised of 10 rats/sex and recovery groups 5 rats/sex as described below:

Group numbers

Dose (mg/kg/day)

Number of Animals

Dose volume (mL/kg)

Terminal sacrifice

G1

Vehicle control*

10M + 10F

10

Day 91

G2

100

10M + 10F

10

Day 91

G3

300

10M + 10F

10

Day 91

G4

1000

10M + 10F

10

Day 91

G1R

Vehicle control*

5M + 5F

10

Day 119

G4R

1000

5M + 5F

10

Day 119

M: Males;  F: Females;  * The vehicle was Olive oil

The vehicle or test item suspensions were not administered to the recovery groups (G1R and G4R) for 28 days following the 90-days dosing period. All rats were observed for clinical signs and mortality daily. Body weights and food consumption were measured during the course of in-life phase of the experiment. The functional observation tests were performed during 13thweek of in-life phase. The blood samples were collected for clinical pathology at termination. All the rats were sacrificed at the end of the treatment for main groups and at the end of recovery groups for recovery groups and subjected to gross examination. The study plan specified organs were collected, weighed and preserved.

Histopathological examination was restricted to the preserved organs from vehicle control (G1) and high dose (G4) group animals (with qualitative assessment of stages of spermatogenesis), as there were no identifiable target organs/tissues in the high dose group rats. In addition, all the gross lesions from all the animals were examined microscopically.

Detailed testicular histopathological examination was conducted in order to identify possible treatment-related effects such as retained spermatids, missing germ cell layers or types, multinucleated giant cells or sloughing of spermatogenic cells into the lumen. Intact epididymis was also examined to include the caput, corpus, and cauda, which was accomplished by evaluation of a longitudinal section. The epididymis was evaluated for possible leukocyte infiltration, change in prevalence of cell types, aberrant cell types, and phagocytosis of sperm.As described in paragraphs 41 to 44 (OECD n°416).

The identity of the Acetalization product between glucose and C16/18 (even numbered)-alcohol was provided by the study Sponsor by a Certificate of Analysis. The correct identity and purity of the test item are the responsibility of the Sponsor. The test item was not authenticated at the test facility.

Summary of Results:  

·        No mortalities were observed in males and females of main toxicity and recovery groups at all dose levels.

·        The salivation (slight to moderate) was observed in both sexes at all the test item treated groups during the course of the treatment period. This clinical sign of salivation was observed at 5-10 minutes post-dose and was persisted up to approximately 45 minutes post-dose only. The incidences of salivation were more in the high dose group rats (1000 mg/kg/day) and these were not observed consistently in same rats.The observed clinical sign of salivation was considered as transient andtest item-related non-adverse effect. There were no other clinical signs observed in any of the test item treated groups.

·        Body weights, net body weight gains and food consumption were unaffected by the treatment at all dose levels.

·        No eye abnormalities were found in the ophthalmological examination conducted at the end of the treatment and recovery period.

·        There were no test item-related neurological abnormalities observed at at the end of the treatment and recovery period.

·        There were no test item-related changes in hematology, coagulation, clinical chemistry, urinalysis, terminal fasting body weights, organ weights/ratios and sperm parameters.

·        There were no test item-related gross and microscopic changes among the doses tested.

Conclusion:

The repeated administration of test item Acetalization product between glucose and C16/18 (even numbered)-alcohol to Wistar rats through oral gavage for 90 consecutive days at the dose levels of 100, 300 and 1000 mg/kg Bwt/day did not cause any toxicological effects on general health, mortality, functional observational battery, growth and food consumption. No test item-related changes were observed in haematology, coagulation, clinical chemistry, urinalysis, terminal fasting body weights, organ weights/ratios and sperm parameters and gross and histopathological changes.The salivation (slight/moderate) which was observed in all rats at all the doseswas considered transient andtest item-related non-adverse effect. Hence, under the test conditions, the evaluated “No Observed Adverse Effect Level (NOAEL)” for test item Acetalization product between glucose and C16/18 (even numbered)-alcohol is 1000 mg/kg Body weight/day under the test conditions and doses employed.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
K1

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Critical effects observed:
not specified
Conclusions:
Effects upon inhalation of the substance were not investigated. This is not required as repeat-dose toxicity was assessed by oral route, the substance is non-volatile (vapour pressure of 6.66 x 10-5 Pa) and the granulometry of the substance (only 0.7% of particles are smaller than 1 mm as the substance is in the form of pellets or flakes) would lead to negligible exposure of terminal airways.
Executive summary:

Effects upon inhalation of the substance were not investigated. This is not required as repeat-dose toxicity was assessed by oral route, the substance is non-volatile (vapour pressure of 6.66 x 10-5 Pa) and the granulometry of the substance (only 0.7% of particles are smaller than 1 mm as the substance is in the form of pellets or flakes) would lead to negligible exposure of terminal airways.

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Critical effects observed:
not specified
Conclusions:
Effects upon repeated skin contact with the substance were not investigated. This is not required as the substance was non-toxic in an acute dermal toxicity study (see 5.2.1.3), repeat-dose toxicity was assessed by oral route and the physicochemical properties (see discussion under 5.1.1) are not in favour of a significant dermal absorption.
Executive summary:

Effects upon repeated skin contact with the substance were not investigated. This is not required as the substance was non-toxic in an acute dermal toxicity study (see 5.2.1.3), repeat-dose toxicity was assessed by oral route and the physicochemical properties (see discussion under 5.1.1) are not in favour of a significant dermal absorption.

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Based on the similarity of chemical structure and physico-chemical properties, there is no reason to expect any differences in toxicokinetic behaviour among the category members. Absorption via the dermal and inhalation route will be limited due to the low water solubility (<1 mg/L), the high logKow (>7.7) and the low vapour pressure (1.19*10 -3 Pa) for acetalization products between glucose and C16/18 alcohol.

By oral route, the alkylpolyglucoside in the UVCBs are hypothesized to undergo hydrolytic deglycosylation, leading to formation of glucose and fatty alcohol. The fatty alcohol part may turn into fatty acids, and then possibly undergo the same metabolism as endogenous fatty acids. The limited toxicity of the substances that were tested after repeated exposure confirms these expectations.

The registered substance included in finished cosmetic products will be applied on skin for rinse-off and leave-on uses.

A repeated patch test was performed on 50 human volunteers at the maximum dose recommended of 5%, no significant dermal reaction was observed according to the protocol followed (HRIPT). A use test was also performed with the registered substance diluted at 5% and applied under normal conditions of use, twice a day for 4 weekks in 21 female volunteers. The product was well tolerated and non comedogenic.

Justification for classification or non-classification

The substance is not classified as a systemic toxic agent