Registration Dossier

Administrative data

Description of key information

Oral (OECD 422) rosin fumarated, rat: NOAEL = 221-228 mg/kg bw/d (males) and 196-292 mg/kg bw/d (females)

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
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
Study period:
2003
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP Study conducted according to guideline protocol tested with the source substance CAS 65997-04-8. Based on the structural similarities and the fact that the target substance is an adduct of the source substance, this study is considered valid for read-across.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
yes
Remarks:
It did not include neurotoxicity screening
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 (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 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):20+/-2
- Humidity (%):50+/-15
- Air changes (per hr): minimum 15
- Photoperiod (hrs dark / hrs light):12/12

IN-LIFE DATES: From: March 31, 2003 To: June 26, 2003
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 mating
Females: commencing 2 weeks prior to mating, then through mating until termination after Day 4 of lactation.
Frequency of treatment:
Continuous
Remarks:
Doses / Concentrations:
0, 1000, 3000, 10000ppm
Basis:
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
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 recorded

BODY 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 time

CLINICAL 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 examinations

HISTOPATHOLOGY: 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.
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 MORTALITY
There were no mortalities.
There was abnormal coloured urine at all treatment levels and soft faecal output at 3000 and 10000ppm

BODY WEIGHT AND WEIGHT GAIN
At 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.

HAEMATOLOGY
At 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 CHEMISTRY
Total bilirubin was significantly increased in both sexes at 10000ppm.

ORGAN WEIGHTS
In 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 PATHOLOGY
All findings were consistent with those normally observed for this age and strain of rat.

HISTOPATHOLOGY: NON-NEOPLASTIC
There 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.

Dose descriptor:
NOEL
Effect level:
1 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No clear effects of toxicity at this level in parental rats.
Dose descriptor:
NOAEL
Effect level:
3 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No clear effects of toxicity at this level in parental rats.
Dose descriptor:
NOEL
Effect level:
3 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: For reproductive parameters which were also determined
Critical effects observed:
not specified

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 3000ppm.

Conclusions:
The NOEL in this study can be determined at 1000ppm for both male and female rats.
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).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
196 mg/kg bw/day
Study duration:
subacute
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

After oral administration of resin acids and rosin acids, fumarated, compds. with triethanolamine hydrolysis of the esterbonds is expected at the low pH in the stomach. Therefore the potentially absorbed compounds would be rosin, rosin fumarated and triethanol amine. After dermal administration no or very limited hydrolysis is expected and the compounds absorbed would be rosin, rosin fumarated and the esters.

Oral

Data on oral repeated dose toxicity by the analogue and hydrolysis product rosin, fumarated were used for the assessment. Rosin, fumarated was administered in the diet to male and female rats at concentrations of 0, 1000 ppm, 3000 ppm and 10000 ppm for approximately 6-7 weeks in a screening study performed according to GLP and OECD 422 (Clubb, 2004). Food consumption and mean body weights were decreased in both sexes at 10000 ppm and 3000 ppm, with high dose animals also showing an increase in total bilirubin (both sexes) and decreased adrenal weight (females only). The lower of these two values will be used as the NOAEL for the repeated dose toxicity. This is considered a scientifically defensible since, apart from poor palatability and associated body weight reduction following exposure to 10000 ppm test substance, no clearly adverse effects were apparent. The NOAEL for repeated dose toxicity of Rosin, fumarated is therefore 221-228 mg/kg bw/d for males and 196-292 mg/kg bw/d for females.

 

Dermal

A subchronic dermal toxicity study with the other metabolic product triethanolamine is available.

In this GLP-study conducted by the National Toxicology Program (NTP, 1999) groups of 10 male and 10 female rats were topically administered 0, 125, 250, 500, or 1000 mg/kg bw/d triethanolamine in acetone or 2000 mg/kg neat triethanolamine, 5 days per week, for 13 weeks. All rats survived to the end of the study. Final mean body weights and weight gains of males and females administered 2000 mg/kg and the mean body weight gain of females administered 1000 mg/kg were significantly less than those of the vehicle controls. Clinical observations included irritation, scaliness, and crustiness of the skin at the site of application for males and females. Males also had discoloration, and two males administered 2000 mg/kg had ulceration at the site of application. Changes in clinical pathology parameters were minor and consistent with inflammation at the site of application. Kidney weights were generally greater in males and females administered 500, 1000, or 2000 mg/kg than in the vehicle controls. Microscopic lesions attributed to triethanolamine administration included acanthosis and inflammation at the site of application, nephropathy in females, and hypertrophy of the pituitary gland pars intermedia in males and females. Although this incidence of nephropathy was increased from low to high dose in female rats, the severity of this lesion did not vary between dose groups. Thus, these effects were regarded as incidental. Based on these data, the NOAEL for systemic effects was found to be 500 mg/kg bw/d for females and 125 mg/kg bw/d for males, while the NOEL for local effects was 250 mg/kg bw/d for females and 125 mg/kg bw/d for males respectively.

The tested substance triethanolamine has a determined dermal absorption potential of 1.85 × 10–2cm/h with a high dermal absorption, higher than that expected for resin acids and rosin acids, fumarated, compds. with triethanolamine (high molecular weight and logPow > 5.8). The internal concentration of triethanol amine in the study above is therefore expected to be much higher than the concentration that could be reached after dermal application of resin acids and rosin acids, fumarated, compds. with triethanolamine (where no hydrolysis is expected and thus dermal exposure to TEA is likely to be minimal).

Discussion

In view of the use of the substance the oral exposure route is not relevant for worker exposure (consumer exposure is not anticipated). Because for the dermal route limited hydrolysis of the substance is expected, the oral study on the rosin fumarated will be taken as starting point for risk assessment. Rosin fumarated has logPow of 2.8 and a molecular weight favourable for absorption, thus representing a worst case compound for absorption compared to the other components. Route-to-route extrapolation will be possible as for rosin fumarated hydrolysis is not applicable. As the maximum amount of TEA that can be absorbed afgter hydrolysis is very small, this substance is not taken into further considerations.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Reliable study on rosin fumarated.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
No inhalation study on the substance or its analogue available. route to route application will be applied

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
No dermal study on the substance or its analogue available. route to route application will be applied

Justification for classification or non-classification

The available data on oral repeated dose toxicity of components of the test substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.