Registration Dossier

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Study conducted according to guideline protocol
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
not applicable
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Rosin, fumarated- Substance type: chemicallty modified UVCB- Physical state: amber solid- Analytical purity: normal commercial sample- Lot/batch No.: TWR01075-15FTOR-9.19.01- Expiration date of the lot/batch: 21 September 2006- Storage condition of test material: in the dark at room temperature.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Charles River (UK) Limited, Margate, Kent, England- Age at study initiation: (P) 6 x wks on arrival;- Weight at study initiation: on arrival (P) Males: 140-169 g; Females: 111-138 g;- Fasting period before study: none- Housing: 2 per cage initially, in polypropylene cages, with stainless steel grid bottoms and mesh tops. A few days prior to pairing for mating, males were transferred to individual cages of similar design. Mated females were transferred to individual solid bottomed cages.- Use of restrainers for preventing ingestion (if dermal): n/a- Diet (e.g. ad libitum): Ad libitum. Rat and Mouse Breeder Diet No. 3 (Expanded Ground) SQC, (Special Diets Services Ltd., Essex, UK)- Water (e.g. ad libitum): Ad libitum, domestic mains water.- Acclimation period: 13 daysENVIRONMENTAL CONDITIONS- Temperature (°C):20+/-2- Humidity (%):50+/-15- Air changes (per hr): minimum 15- Photoperiod (hrs dark / hrs light):12/12IN-LIFE DATES: From: March 31, 2003 To: June 26, 2003

Administration / exposure

Route of administration:
oral: feed
Vehicle:
other: diet
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:DIET PREPARATION- Rate of preparation of diet (frequency): Weekly, and used within 15 days of preparation- Mixing appropriate amounts with (Type of food): Rat and Mouse Breeder Diet No. 3 (Expanded Ground) SQC - Storage temperature of food: No data.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Diet formulations were analysed on 2 occasions during the study treatment period. Analysis of formulation diets was undertaken with regard to concentration and homogeneity. Diet prepared for Week 1 and Week 4 of treatment was sampled. On each occasion, triplicate samples were taken from each formulated diet containing test item and from the control diet. Samples were analysed by previously validated method.
Duration of treatment / exposure:
Males: at least 4 weeks overall - starting 2 weeks prior to matingFemales: commencing 2 weeks prior to mating, then through mating until termination after Day 4 of lactation.
Frequency of treatment:
Continuous
Doses / concentrations
Remarks:
Doses / Concentrations:0, 1000, 3000, 10000ppmBasis:nominal in diet
No. of animals per sex per dose:
10
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: Dose levels selected and agreed with Sponsor, following evaluation of existing toxicological data. This included data from a one week dose range finding study in rats carried out under a separate contract and project number at the laboratory.- Rationale for animal assignment (if not random): random- Rationale for selecting satellite groups:n/a- Post-exposure recovery period in satellite groups:n/a- Section schedule rationale (if not random):n/a
Positive control:
none

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes- Time schedule: Daily for viability early in the morning and again as late as possible on each day.DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: Daily. Nature, onset, duration and intensity of any signs were recordedBODY WEIGHT: Yes- Time schedule for examinations: Males - once during the week prior to the commencement of dosing and once weekly thereafter until termination.Females - once during the week prior to the commencement of dosing, and weekly thereafter until the start of the mating period. Then on Day 0 of gestation followed by Days 7, 14, and 20 of gestation, and then Days 1 and 4 of lactation (where Day 0 of lactation is day of parturition).FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes - Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes FOOD EFFICIENCY:- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No OPHTHALMOSCOPIC EXAMINATION: No- Time schedule for examinations:- Dose groups that were examined:HAEMATOLOGY: Yes - Time schedule for collection of blood: Week 5 of dosing for males and Day 6 lactation for females- Anaesthetic used for blood collection: No. Taken from lateral tail vein.- Animals fasted: No - How many animals: Initially 5 males and 5 females from each dose group. Then one additional sample was taken for female controls to ensure a total of 5 samples. Additiona samples were taken from 2 high dose females on Day 7 of lactation owing to problems with obtaining sufficient samples for analysis. - Parameters checked: Haematology: Haemoglobin, RBC count, Haematocrit, WBC count, MCV, mean cell haemoglobin, mean cell haemoglobin concentration, platelets, differential WBC count, neutrophils, lymphocytes, monocytes, eosinophils, basophils, large unclassified cells.Coagulation: Prothrombin timeCLINICAL CHEMISTRY: Yes - Time schedule for collection of blood: Week 5 of dosing for males and Day 6 lactation for females- Animals fasted: No - How many animals: Initially 5 males and 5 females from each dose group. Then one additional sample was taken for female controls to ensure a total of 5 samples. Additiona samples were taken from 2 high dose females on Day 7 of lactation owing to problems with obtaining sufficient samples for analysis. - Parameters checked: urea, glucose, aspartate aminotransferase, alanine aminotransferase, sodium potassium, chloride, total protein, albumin, A:G ration, creatinine, calcium, phosphate, total bilirubin, cholesterol, alkaline phosphatase. URINALYSIS: No- Time schedule for collection of urine:- Metabolism cages used for collection of urine: Yes / No / No data- Animals fasted: Yes / No / No data- Parameters checked in table [No.?] were examined.NEUROBEHAVIOURAL EXAMINATION: No - Time schedule for examinations:- Dose groups that were examined:- Battery of functions tested: sensory activity / grip strength / motor activity / other:OTHER: no data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes - Gross necropsy consisted of external and internal examinationsHISTOPATHOLOGY: Yes -The tissues indicated in Table 1 were prepared for microscopic examination and weighed, respectively. Histological examination was conducted on control and high dose animals only.
Other examinations:
Pups were examined for externally visible abnormalities and discarded following examination.The following parameters were examined in F1 offspring:number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, other: any deficiencies in maternal care were recorded.
Statistics:
Body weight, food consumption (prior to mating for females), haematology and clinical chemistry were statistically analysed for homogeneity of variance using the 'F-max' test. If the group variances appeared homogeneous, a parametric ANOVA was used and pairwise comparisons made via Student's t-test using Fisher's F-protected LSD. If the variances were heterogeneous, log or square root transformations were used in an attempt to stabilise the variances. If the variances remained heterogenous then Kruskal-Wallis ANOVA was used.Organ weights were analysed as above and by analysis of covariance (ANCOVA) using terminal kill body weight as covariate.Histology incidence data were analysed using Fisher's Exact Probability Test.All tests were two-sided and performed at the 5% significance level.

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):
effects observed, treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITYThere were no mortalities.There was abnormal coloured urine at all treatment levels and soft faecal output at 3000 and 10000ppmBODY WEIGHT AND WEIGHT GAINAt 3000 and 10000ppm there was a decrease in mean body weight gain in both sexes. There were no effects in animals treated at 1000ppm.FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)Mean food consumption was reduced for both sexes at 3000 and 1000ppm. At 10000ppm in both sexes, and among males treated at 3000ppm, the achieved intake in the first week of treatment was lower than in the second week. Among females treated at 10000ppm there was a decreased intake over Days 0-7 and 14-20 of gestation and also during lactation.HAEMATOLOGYAt 10000ppm males there were slight decreases in haemoglobin, red blood cell count and haematocrit. A decrease in monocytes in males treated at 3000ppm was statistically significant but since it was not associated with dose it was considered to be incidental.Neutrophil counts in females were subject to a high degree of variability and as such these findings were considered to be coincidental. CLINICAL CHEMISTRYTotal bilirubin was significantly increased in both sexes at 10000ppm.ORGAN WEIGHTSIn males treated at 10000ppm there was a slight decrease in absolute lung and spleen weights, attaining statistical significance, but following covariance analysis these differences from control were no longer evident.In females, mean kidney, spleen, adrenal gland, ovary and uterus weights were significantly reduced at 10000ppm. Following covariance analysis, kidney, spleen and uterus weights were essentially similar to controls, but a decrease in adrenal weight was still evident.Mean salivary gland weights in females also achieved statistical significance in all treatment groups, but following adjustment for the lower body weight by analysis of covariance these differences were no longer apparent.GROSS PATHOLOGYAll findings were consistent with those normally observed for this age and strain of rat.HISTOPATHOLOGY: NON-NEOPLASTICThere were no obvious effects of treatment on the type or incidence of the histology findings.Thymic atrophy in females treated at 10000ppm was increased compared to controls, but given the reduced body weight at this level, during pregnancy and lactation, the findings may reflect the physiological status of the animals and may not reflect an effect of treatment.

Effect levels

Dose descriptor:
NOAEL
Effect level:
3 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No evidence of toxicity. Equivalent to received doses of 221-288 mg/kg bw/d for males, and 196-292 mg/kg bw/d for females.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Reproductive indices (fertility index, gestation index and birth index) and offspring viability indices (live birth index, viability index) were determined. At 10000ppm there was a slight increase in the number of nights to a positive mating sign, and in the duration of gestation. Additionally, the mean number of implants per pregnancy was slightly decreased with a subsequent reduction in litter size and litter weight. There were no obvious effects of treatment on mating performance, duration of gestation, litter size and pup weight in animals treated at 1000 and 3000 ppm.

Applicant's summary and conclusion

Conclusions:
Rosin fumarated was administered in the diet to rats at concentrations of 0, 1000ppm (males 72 -89 mg/kg bw/d; females 79 -108 mg/kg bw/d), 3000 ppm (males 221 -288 mg/kg bw/d; females 196 -292 mg/kg bw/d), and 10000 ppm (males 651 -889 mg/kg bw/d; females 449 -995 mg/kg bw/d). Under the conditions of this study, parental toxicity was exhibited at levels of 3000 and 10000 ppm, with a decrease in mean body weight at both doses and sexes, an increase in total bilirubin in both sexes at 10000 ppm and decrease in adrenal gland weight in females at 10000 ppm, but there were no clear effects of toxicity at 1000 ppm. Therefore, the parental NOEL was considered to be 1000 ppm (males 72 -97 mg/kg bw/d; females 79 -108 mg/kg bw/d). The effects seen do not constitute 'serious damage to health' at a dose level requiring classification as R48 'Danger of serious damage to health by prolonged exposure' or 'significant toxic effects' at a dose level requiring classification under the CLP Regulation (EU Regulation 1272/2008). Therefore, the test material is not classified according to UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) or EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008. This substance is not classified in the Annex I of Directive 67/548/EEC.
Executive summary:

This is a repeat dose toxicity study conducted according to guideline protocol with the objective of providing initial information on possible effects on reproduction and/or development in rats. Rosin fumarated was administered in the diet to rats at concentrations of 0, 1000ppm (males 72 -89 mg/kg bw/d; females 79 -108 mg/kg bw/d), 3000 ppm (males 221 -288 mg/kg bw/d; females 196 -292 mg/kg bw/d), and 10000 ppm (males 651 -889 mg/kg bw/d; females 449 -995 mg/kg bw/d). The males were treated for 2 weeks prior to mating, through until necropsy after 4 weeks of treatment. The females were treated for 2 weeks prior to mating, then through mating, gestation and until termination on at least Day 4 of lactation. Under the conditions of this study, parental toxicity was exhibited at levels of 3000 and 10000 ppm, with a decrease in mean body weight at both doses and sexes, an increase in total bilirubin in both sexes at 10000 ppm and decrease in adrenal gland weight in females at 10000 ppm, but there were no clear effects of toxicity at 1000 ppm. Therefore, the parental NOEL was considered to be 1000 ppm (males 72 -97 mg/kg bw/d; females 79 -108 mg/kg bw/d). For reproductive parameters the NOEL was considered to be 3000 ppm (females: 196 -268 mg/kg bw/d).