L-gluco-Octitol, 1,5-anhydro-6,8-dideoxy-, (7.xi.)-

Endpoint summary

Description of key information

In a 28-day repeated dose oral toxicity study conducted according to the OECD Guideline 407 in rats, the NOEL was 1000 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 October 2004 - 21 January 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted according to OECD Guideline 408 with minor deviation: some fluctuations of temperature and humidity outside the range were observed on a day

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

some fluctuations of temperature and humidity outside the range were observed on a day

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories France, L’Arbresle, France
- Age at study initiation: Approximately 6 weeks
- Weight at study initiation: Males: 198-228 g (mean: 213 g); females: 133-174 g (mean: 155 g)
- Housing: Housed in groups of 2/sex in suspended wire-mesh cages
- Diet: SSNIFF R/M-H pelleted maintenance diet (SSNIFF Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: Tap water (filtered with a 0.22 μm filter), ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 50 ± 20 %
- Air changes: Approximately 12 cycles/h of filtered, non-recycled air
- Photoperiod: 12 h dark / 12 h light

Route of administration:

oral: gavage

Vehicle:

other: 60/40 (w/w) purified water/propylene glycol

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
- Test item was diluted with the required quantity of vehicle in order to achieve the desired concentrations and then homogenized using a magnetic stirrer.
- Test item dosage forms were prepared for up to 9 days and stored at +4 °C. They were used each day within a maximum of 6 h storage at room temperature.

VEHICLE
- Concentration in vehicle: 20, 60 and 200 mg/mL
- Dose volume administered (for vehicle and treatment groups): 5 mL/kg bw/day
- Lot/batch no. of propylene glycol: S21645-174, S21645-284, S21645-334 and U10221
- Source of propylene glycol: Aldrich (Saint-Quentin-Fallavier, France)
- Source of purified water: Obtained by reverse osmosis using a Milli-Ro 8 plus apparatus (Millipore SA, Saint-Quentin en Yvelines, France)

STABILITY:
- Two dosage forms were prepared at 10 and 200 mg/mL of test item to cover all the concentrations intended for use in this study.
- To evaluate the stability, each dosage form was sampled (in duplicate) after 0 (just after preparation), 6 h storage at room temperature, then after 4 and 9 days storage at +4 °C. The aliquots sampled on Day 4 were analysed immediately after sampling.
- Results of the analyses demonstrated the satisfactory stability of the two dosage forms investigated (10 and 200 mg/mL) over a 6 h period at room temperature and a 9-day period at +4 °C (protected from light).

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

- Concentration of test item in samples taken from each dosage form (including the control) prepared for use in Weeks 1, 4, 8 and 13 was determined.
- Results of analysis: Satisfactory agreement was observed between the nominal and actual concentrations of the test item in the administered dosage forms analysed as the deviations from nominal concentration were in an acceptable range of ± 10 %.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Once daily

Remarks:

Doses / Concentrations:
100, 300 and 1000 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels were selected on the basis of the results of a previous study (Study No. 28461 TSR) in which the test item was given orally to Sprague-Dawley rats for 2 weeks at dose-levels of 100, 300 or 1000 mg/kg bw/day. In absence of signs of toxicity at the high dose-level, 1000 mg/kg bw/day was selected as the highest dose-level.
- Rationale for animal assignment: Animals were selected according to body weight and clinical condition and grouped (by sex) according to a computerised stratification procedure, so that the average body weight of each group was similar.

Positive control:

Not applicable

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
Time schedule:
- Mortality and morbidity: Twice a day
- Clinical signs: Once daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once before the beginning of the treatment period and then once a week until the end of the study

BODY WEIGHT: Yes
- Time schedule for examinations: Once before group allocation, on the first day of treatment, then twice a week until the end of the study and before sacrifice.

FOOD CONSUMPTION:
- Food consumption for each animal was recorded once a week and mean daily diet consumption was calculated as g food/animal/day

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Ophthalmological examinations were performed on all animals, before the beginning of the treatment period and on animals of the control and high-dose groups on one occasion at the end of the treatment period.

HAEMATOLOGY AND CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood samples were taken from the orbital sinus of all the animals at the end of the treatment period.
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes
- Parameters checked (haematology): Hemoglobin, erythrocyte, mean cell volume, packed cell volume, mean cell hemoglobin concentration (MCHC), mean cell hemoglobin (MCH), thrombocytes, leucocytes, differential white cell count (neutrophils, lymphocytes, monocytes, eosinophils, basophils), reticulocytes, prothrombin time, activated partial thromboplastin time and fibrinogen
- Parameters checked (clinical chemistry): Sodium (Na+), potassium (K+), chloride (Cl-), calcium (Ca++), inorganic phosphorus, urea, creatinine, glucose, total bilirubin, direct bilirubin, total protein, albumin, albumin/globulin ratio, total cholesterol, triglycerides, alkaline phosphatase (ALP), aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT)

URINALYSIS: Yes,
- Time schedule: Urinalysis was performed in all animals at the end of the treatment period.
- Metabolism cages used for collection of urine: Yes, animals were individually placed in metabolism cages for an overnight period of at least 14 h.
- Animals fasted: Yes
- Parameters checked: Appearance, color, volume, specific gravity, pH, protein, glucose, ketones, bilirubin, nitrites, blood, urobilinogen, cytology of sediment (leucocytes, erythrocytes, cylinders, magnesium ammonium phosphate crystals, calcium phosphate crystals, calcium oxalate crystals and cells)

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Each animal was observed once in Week 12, in the cage, in the hand and in the standard arena.
Battery of functions tested:
- In the cage: ‘Touch escape’ or ease of removal from the cage.
- In the hand: Fur appearance, salivation, lachrymation, piloerection, exophthalmia, reactivity to handling, pupil size (presence of myosis or mydriasis),
- In the standard arena (two-minute recording): Grooming, palpebral closure, defecation and urination counts, tremors, twitches, convulsions, gait, arousal (hypo- and hyper-activity), posture, stereotypy and breathing, ataxia, hypotonia.
In addition, the following parameters, reflexes and responses were recorded:
- touch response, forelimb grip strength, pupil reflex, visual stimulus, auditory startle reflex, tail pinch response, righting reflex, landing foot splay, rectal temperature at the end of observation.
- Motor activity of each animal was also measured by automated infra-red sensor equipment over a 1 h period.

Sacrifice and pathology:

- GROSS PATHOLOGY: On completion of the treatment period, after at least 14 h fasting, all surviving animals were anesthetized with carbon dioxide and then sacrificed by exsanguinations. A complete macroscopic post-mortem examination was performed on all study animals. This included examination of the external surfaces, all orifices, the cranial cavity, the external surfaces of the brain and spinal cord, the thoracic, abdominal and pelvic cavities with their associated organs and tissues and the neck with its associated organs and tissues.
- ORGAN WEIGHTS: Body weight of each animal was recorded before sacrifice and the organs specified in the table 7.5.1/1 were weighed wet as soon as possible after dissection. The ratio of organ weight to body weight (recorded immediately before sacrifice) was calculated.
- HISTOPATHOLOGY: For all animals, the tissues specified in the table 7.5.1/1 were preserved in 10 % buffered formalin (except for the eyes and Harderian glands which were fixed in Davidson's fixative, and the testes and epididymides which were preserved in Bouin's fluid). All tissues required for microscopic examination were embedded in paraffin wax, sectioned at a thickness of approximately 4 µm and stained with hematoxylin-eosin (except testes and epididymides which were stained with hematoxylin/PAS).
Microscopic examination was performed on:
- all tissues listed in the table 7.5.1/1 for animals in control and 1000 mg/kg bw/day treatment groups sacrificed at the end of the treatment period and for all animals that died
- all macroscopic lesions from all the animals in 100 and 300 mg/kg bw/day treatment groups sacrificed on completion of the treatment period

Other examinations:

None

Statistics:

Refer figure 7.5.1/1 for the details of statistical analysis

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

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY:
- No unscheduled deaths occurred during the study.
- No clinical signs related to the test item treatment were observed.
- Alopecia of forelimbs, thinning of hair, scabs on head and hyperactivity were considered not to be of toxicological importance as they were noted at a low incidence, without clear dose-relationships, and/or these signs are commonly recorded among rats of this strain and age.

BODY WEIGHT AND WEIGHT GAIN:
- Body weight gain of test item-treated animals was comparable to that of the controls.

FOOD CONSUMPTION:
- Food consumption of treated animals was comparable to that of the controls throughout the study.

OPHTHALMOSCOPIC EXAMINATION:
- There were no ocular findings which could be related to the test item treatment.
- An increase in the corneal thickness was observed in one male and one female given 1000 mg/kg bw/day. As this finding was recorded at a low incidence, a relationship to treatment with the test item was doubtful and it was considered to have occurred by chance.

HAEMATOLOGY:
- No differences of toxicological importance on hematological parameters noted between test item-treated and control animals.
- Few minor statistically significant differences were considered not to be biologically significant, as the observed changes were not dose-related, all individual values were within historical data range [for white blood cell in males (7.41-17.09 G/L); for platelet in females (479-1326 G/L) and/or were probably due to high control values].

CLINICAL CHEMISTRY:
- No differences of toxicological importance were noted between test item-treated and control animals among the blood biochemical parameters.
- Increase in inorganic phosphorus recorded in the female group treated at 1000 mg/kg bw/day (1.79 vs. 1.53 mmol/L, +17 %) reached statistical significance (p<0.05). All individual values were within historical data range (1.33-2.45 mmol/L). In the absence of a clear dose-relationship, this very slight change was considered to bear no toxicological importance.

URINALYSIS:
- There were no changes in the urinary parameters evaluated qualitatively or quantitatively that could be attributed to treatment with the test item.

NEUROBEHAVIOUR:
- There were no perturbations of the autonomic or physiological functions in any test item-treated animals.
- Motor activity of both males and females measured over a 60-minute period was unaffected by treatment.

ORGAN WEIGHTS:
- No treatment-related effects were noted for organ weights at the end of treatment period.
- Few differences noted between test item-treated and control groups were slight, not dose-related, of opposing trend in the different groups and sexes and were without relevant histopathological findings in the organs examined microscopically, and therefore they were considered to be of no toxicological importance.

GROSS PATHOLOGY AND HISTOPATHOLOGY:
- No treatment-related necropsy or microscopic findings were noted.
- All necropsy or microscopic findings reported at the end of treatment period were those which are commonly occurring in this strain of rat kept under laboratory conditions and were considered to be of no toxicological importance.

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related effects on clinical signs; mortality; body weight; food consumption; ophthalmoscopic examination; haematology; clinical chemistry; urinalysis; gross pathology; organ weights and histopathology

Critical effects observed:

not specified

None

Conclusions:

Under the test conditions, the No Observed Effect Level (NOEL) of test item was considered to be 1000 mg/kg bw/day in Sprague-Dawley rats.

Executive summary:

In a 90-days repeated dose oral toxicity study conducted according to the OECD Guideline 408 and in compliance with GLP, test item was administered daily by oral gavage to groups of Sprague-Dawley, Crl CD® (SD) IGS BR rats (10/sex/dose) at the dose-levels of 0 (vehicle), 100, 300 or 1000 mg/kg bw/day in 60/40 (w/w) purified water/propylene glycol. Examinations during the study included: mortality, clinical signs, functional observation battery (including motor activity), body weight change, food consumption, ophthalmology, hematology, blood chemistry, urinalysis, organ weights, macroscopic examination and histopathology.

No deaths occurred and no major clinical signs were observed during the study. Body weight gains and food consumption were not affected by treatment with the test item. No treatment-related changes were noted in the hematological parameters, blood biochemistry, urinalysis, ophthalmology and neurotoxicological parameters at any dose-level. No treatment-related effects were noted on organ weights or necropsy and microscopic findings.

Under the test conditions, the No Observed Effect Level (NOEL) of test item was considered to be 1000 mg/kg bw/day in Sprague-Dawley rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information as a whole meets the tonnage driven data requirement of REACH. Moreover, reliability and consistency are observed across the different studies.

Additional information

In a 28-day repeated dose oral toxicity study conducted according to the OECD Guideline 407 and in compliance with GLP, MEXORYL SBF was administered daily by oral gavage to groups of Sprague-Dawley rats (5/sex/dose) at the dose-levels of 0 (vehicle), 150, 450 or 1000 mg/kg bw/day in purified water. No treatment-related changes were noted in all the parameters studied therefore the NOEL was 1000 mg/ kg bw/day.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Key study performed on Mexoryl SBF in compliance with OECD Guideline 407 and GLP.

Justification for classification or non-classification

As the NOEL identified in a subacute repeated dose toxicity study is at least 1000 mg/kg bw/day, there is no need for classification of MEXORYL SBF according to the Annex VI to the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272 -2008).