C12-14 (even numbered) alkyl glycosides, oligomeric and C12-14 (even numbered) alkyl glycosides, oligomeric, carboxymethyl ethers, sodium salts

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-06-24 to 2003-06-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline Study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

21 September 1998

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Italy s.r.l; 33049 San Pietro al Natisone (UD); Italy
- Age at study initiation: 27 - 29 days
- Weight at study initiation: males: 89 - 95 g; females. 80 - 95 g (nulluparous and non-pregnant)
- Fasting period before study: no
- Housing: polycarbonat cages, 59x38.5x20 cm with stainless steel mesh lid floor (Code 1352 G, Techniplast Gazzeda S.a.r.l, Buggugiate, Varese); 5 animals of one sex per cage;
- Diet: laboratory rodent pelleted diet; Altromin MT, Altromin D-32770 Lage, Postfach 1120, Germany; ad libitum
- Water: drinking water ad libitum
- Acclimation period: overall 17 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 2 °C
- Humidity: 55 +/- 15 %
- Air changes: approximately 15 to 20 air changes per hour
- Photoperiod: 12 hours light/12 hours dark per day

IN-LIFE DATES: From: 2002-06-24 To: 2002-10-28

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Required amount of test item was dissolved in distilled water. Formulations were prepared daily.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

The required amount of the test item was dissolved in the vehicle (distilled water). The formulations were prepared daily (concentrations of 2.5, 7.5 and 22.5 mg/mL).

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 50, 150, 450 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

10 males, 10 females per dose in the treatment groups, 5 animals per sex and per dose in the recovery groups

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based on the results of a range finding pre-test
- Rationale for animal assignment: randomizing
- Rationale for selecting satellite groups: control and high dose group included 5 additional animals per sex
- Post-exposure recovery period in satellite groups: 4 weeks
- Ratiuonale for animals assignment: randomizing:

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: All animals were weighed on the day of allocation to treatment groups , on the day of that treatment commenced, weekly thereafter and just prior to necropsy.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: All animals prior to commencement of the study, during week 12 of treatment
- Dose groups that were examined: Control and high dose animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Once during week 13 of the treatment period, once during week 4 of the recovery period;
- Anaesthetic used for blood collection: Yes (light ether anaesthesia)
- Animals fasted: Yes
- How many animals: all male and female animals
- Parameters checked: Haematocrit, haemoglobin, red blood cell count, reticulocyte count, mean red blood cell volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, white blood cell count, differential leucocyte count (neutrophils, lymphocytest, eosinophils, basophils monocytes, large unstained cells), abnormalities of the blood film, platelets, prothrombin time.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Once during week 13 of the treatment period, once during week 4 of the recovery period;
- Animals fasted: Yes
- How many animals: all male and female animals
- Parameters checked: alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, urea, blood urea nitrogen, creatinine, glucose, albumin total bilirubin, total cholesterol, total protein sodium, potassium, calcium, chloride

URINALYSIS: Yes
- Time schedule for collection of urine: Once during week 13 of the treatment period, once during week 4 of the recovery period
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked: Appearance, volume, specific gravity, pH, protein, total reducing substances, glucose, ketones, bilirubin, urobilinogen, blood, sediment

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Once before commencement of treatment and one week thereafter
- Dose groups that were examined: each animal
- Battery of functions tested: sensory activity / grip strength / motor activity / other: gait, tonic movements, clonic movements

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Organs examined in necropsy and histopathology:
Abnormalities, adrenal glands, aorta, bone marrow (from sternum), brain, caecum, colon, duodenum, eyes, epididymides, femur with joint, heart, ileum (including Peyer's patches), jejeunum, kidneys, liver, lungs, lymph nodes-mesenteric, lymph nodes-cervical, mammary area, oesophagus, ovaries, pancreas, parathyroid gland; pituitary gland, prostate gland, rectum, salivary glands, sciatic nerve, seminal vesicles, skeletal muscle, skin, spinal column, spinal cord, spleen, stomach/forestomach, testes, thymus (when present), thyroid, trachea urinary bladder, uterus

Statistics:

For continuous variables the significance of the differences amongst groups was assessed by analysis of variance. Differences between each treated group and the control group were assessed by Dunnett's test using a pooled error variance. The homogeneity of the data was verified by Bartlett's test before Dunnett's test. If data were found to be inhomogeneous a Modified t test (Cochran and Cox) was applied. The mean values, standard deviations and statistical analysis were calculated from the actual values in the computer without rounding off. A statistical analysis of histopathological findings was not carried out as indicated in the protocol, and, therefore, this is a deviation from protocol.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

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:

effects observed, treatment-related

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY: No mortality occurred during the study. No significant daily post-dose observations were noted during the study. Detailed clinical signs with neurotoxicity assessment did not show any signs which could be correlated to the treatment with the test item.

BODY WEIGHT AND WEIGHT GAIN: No changes of toxicological significance were observed in body weight.

OPHTHALMOSCOPIC EXAMINATION: No significant findings were seen at the ophthalmic examination performed at the end of the study.

HAEMATOLOGY: No changes were seen in haematological parameters.

CLINICAL CHEMISTRY: a statistically significant increase in alkaline phosphatase was observed in high dose females at the end of the treatment period. This change, within the range of historical control data, was no longer present at the end of the recovery period. No other significant changes were noted.

URINALYSIS: An increase in urine volume, statistically significant in all treated males and high dose females, was observed at the end of treatment. This difference, within the range of historical control data, was no longer evident at the end of the recovery period.

NEUROBEHAVIOUR: Detailed clinical signs with neurotoxicity assessment did not show any signs which could be correlated to the treatment with the test item.

ORGAN WEIGHTS: No significant variations were observed in the weights of the organs.

GROSS PATHOLOGY: No macroscopic finding was described that could be considered correlated with the administration of the test item.

HISTOPATHOLOGY: NON-NEOPLASTIC: Keratinised stomach acanthosis was significantly greater than background degree in two animals from the high dose group (450 mg/kg bw/day). At this incidence this lesion is not considered treatment related.

OTHER FINDINGS: other occasional findings are those that are commonplace in the tissues of young laboratory rats and none are related to treatment.

Effect levels

Dose descriptor:

NOEL

Effect level:

450 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The NOEL in this study was 450 mg/gk bw.

Executive summary:

The test substance has been investigated with regard to its toxicity after repeated exposure in a subchronic study in accordance with OECD guideline 408 in male and female SD rats. 10 animals of either sex received vehicle, 50, 150, or 450 mg/kg bw for 90 consecutive days. A recovery high dose group was left without treatment for another 14 days. No effects were observed on body weights, food consumption, organ weights, macroscopic examination, functional observational battery, hematology and histopathology. Urine volume and plasma AP was increased in both sexes but the effects were reversible in the recovery group, the AP values were still in the range of the historical controls and none of the effects was accompanied by corresponding organ/clinical effects. Therefore they were considered to be toxicologically and biologically not relevant. The NOEL was 450 mg/kg bw.