Registration Dossier

Registration Dossier

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Administrative data

Description of key information

No repeated dose toxicity data are available for the registration substance therefore this endpoint is addressed by a weight of evidence approach for relevant groups of constituents present in the substance.

In a 90-day repeated dose toxicity study (Hepburn et al., 1998) conducted using a protocol equivalent to OECD 408 and in compliance with GLP, administration of phytosterol in the diet of Wistar rats for 90 days did not lead to any adverse effects up to dietary concentrations of 8.1% phytosterol ester (equivalent to 6600 mg/kg bw/day; or 4100 mg/kg bw/day phytosterol). Therefore, the NOAEL from this test was 6600 mg/kg bw/day phytosterol ester.

In a 90-day oral repeated dose toxicity test (Envigo, 2017) conducted to OECD 408 and in compliance with GLP with tall oil rosin, the NOAEL was concluded to be 5000 ppm (equivalent to a mean achieved dose of 335.2 mg/kg bw/day for males and 401.2 mg/kg bw/day for females) based on reduced body weight gain in the highest dose group. No other adverse effects were observed.

In a supporting Combined Repeated Dose Toxicity Study with Reproduction/Developmental Toxicity Screening Test conducted according to OECD 422 and in compliance with GLP for distilled tall oil, the reported NOEL was 1000 ppm (approximately 100 mg/kg bw/day) for both male and female rats. The NOEL was based on effects observed in food consumption, body weight gain, organ weights and changes in clinical biochemistry parameters at 20000 ppm and changes in organ weights and clinical biochemistry parameters at 5000 ppm. No effects were observed at 1000 ppm. In conclusion, under the conditions of this study, the parental NOEL was reported to be 1000 ppm. However, the effects on the liver are considered (by the author the study summary) to be adaptive rather than adverse, and decreased food consumption and weight gain were most likely to be due to palatability of the test diet, rather than toxicity. Therefore, the overall NOAEL for general toxicity was 5000 ppm tall oil (equivalent to approximately 423 mg/kg bw/day for males and 483 mg/kg bw/day for females; the highest dose tested).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
No data
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes
Specific details on test material used for the study:
This study used sterol/fatty acid esters as test material, since this is the chemical form in which they are extracted from plants
Species:
rat
Strain:
Wistar
Remarks:
Alpk:APfSD
Details on species / strain selection:
Strain used as the test lab had substantial background data available for it.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Rodent Breeding Unit of Zeneca Pharmaceuticals (Alderley Park, Macclesfield, IJK)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: approximately 6 weeks
- Weight at study initiation: Approximately 145-190g
- Fasting period before study:
- Housing:
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: Up to 2 weeks

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 40-70
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To:
Route of administration:
oral: feed
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): The phytosterol ester was added to the diet (modified form of the AIN (American lnstitute of Nutrition) diet) to give concentrations of 0.16, 1.6,.3.2 and 8.1% (w/w) equivalent to phytosterol ester concentrations of 0.1, 1.0, 2.0 and 5.0% (w/w), respectively. The dry ingredients were mixed in a mixer for approximately 1 minute. The phytosterol ester was then mixed with the maize oil component of the diet using a blender. The oil mixture was then added to the dry ingredients and the complete diet
mixed for 6 minutes.
- Storage temperature of food: A 20-day stability trial conducted at both ambient temperature (19-25°C) and refrigerator temperature (0-4°C) showed PE to be stable in the diet. It is not clear which of these temperatures was used for storage.

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to the start of the study the homogeneity and stability of phytosterol ester in the experimental diets was established. The homogeneity of three dietary phytosterol ester concentrations was determined (0.1, 1.0 and 10% w/w) to cover the range of dose levels anticipated for the feeding study. Each dietary dose level was determined by taking five separate samples for analysis from the following positions in the mixing bowl: top centre, middle centre, bottom centre, left centre, right centre). The diets were analysed on the day of preparation (from the homogeneity samples) which also constitute day 0 of the stability trial.

Four additional samples were taken from each diet; two were stored at ambient temperature (19-25°C) and two were stored refrigerated (0-4°C). A pot from each storage temperature was then analysed on day 13 or day 20 to determine the concentration of phytosterol ester. In addition, the actual achieved concentration of phytosterol ester used in the feeding study was determined by analysis of all dose levels for diets prepared for use in week 1, 7 and 13. In each case the diet samples were extracted with petroleum spirit and analysed by HPLC with UV detection at 210 nm and quantitation was achieved by comparison with standards of known concentrations.

Recovery data showed the method of analysis to be acceptable for levels of phytosterol ester in the diet in the range 0.1% to 10% (w/w) and indicated an error bar of ±20% for the 0.1% level and ±15% for the 1.0% and 10% levels. Homogeneity of mixing the diets was demonstrated, within experimental error, for diet samples prepared at 0.1%, 1.0% and 10% (w/w). A 20-day stability trial conducted at both ambient temperature (19 - 25°C) and refrigerated temperature (0-4°C) showed phytosterol esters to be stable in the diet. The actual concentrations of phytosterol esters in the three batches of diet prepared at week 1, 7 and 13 were within ±12% of the nominal concentrations and within the accepted experimental error of the analytical method used.
Duration of treatment / exposure:
90 days
Frequency of treatment:
Continuous in diet
Dose / conc.:
0.16 other: % (w/w) phytosterol ester
Remarks:
Equivalent to phytosterol concentration of 0.1% (w/w)
Dose / conc.:
1.6 other: % (w/w) phytosterol ester
Remarks:
Equivalent to phytosterol concentration of 1.0% (w/w)
Dose / conc.:
3.2 other: % (w/w) phytosterol ester
Remarks:
Equivalent to phytosterol concentration of 2.0% (w/w)
Dose / conc.:
8.1 other: % (w/w) phytosterol ester
Remarks:
Equivalent to phytosterol concentration of 5.0 % (w/w)
Equivalent to phytosterol dose of 4100 mg/kg bw/day and phytosterol ester dose of 6600 mg/kg bw/day
No. of animals per sex per dose:
20
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale:
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: No satellite groups
- Post-exposure recovery period in satellite groups: No post exposure group
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: On day 1 then weekly thereafter (at the same time as body weight measurements).

BODY WEIGHT: Yes
- Time schedule for examinations: On day 1 then weekly thereafter.

FOOD CONSUMPTION AND COMPOUND INTAKE:
- 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: yes

WATER CONSUMPTION: Yes
- Time schedule for examinations: Continuous

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: An ophthalmoscopic examination of all the animals was conducted prior to the start of the study. The eyes of rats from the control and high dose groups were also examined during week 13. The examination was carried out using a Fison's binocular indirect ophthalmoscope after instillation of 0.5% (w/w) tropicamide into the eyes to dilate the pupils.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the end of the 90 day feeding period.
- Anaesthetic used for blood collection: Yes (halothane)
- Animals fasted: Not specified
- How many animals: All animals
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At the end of the 90 day feeding period.
- Animals fasted: Not specified
- How many animals: All animals
- Parameters checked in table [No.1] were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table 2)

HISTOPATHOLOGY: Yes (see table 2)
Statistics:
Body weights were considered by analysis of covariance on initial (week 1) body weight, separately for males and females. Weekly food consumption, food utilisation and weekly water consumption were considered by analysis of variance, separately for males and females.
Haematology and clinical chemistry were considered by analysis of variance. Male and female data were analysed together and results examined to determine whether any differences between control and treated groups were consistent between sexes.

Organ weights were considered by analysis of variance and analysis of covariance on final body w eight, separately for males and females. All data were evaluated using the GLM procedure in SAS version 6.12 (1989). Least squares means for each group were calculated using the LSMEAN option in SAS PROC GLM. Analysis of variance and covariance allowed for the replicate structure of the study design. Unbiased estimates of differences from control were provided by the difference between each treatment qroup least-squares mean and the control group least-squares mean.

Differences from the control were tested statistically by comparing each treatment group least squares mean with the control group least squares mean using a two sided Student's t-test based on the error mean square in the analysis.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Description (incidence):
Three rats of the low dose group were killed for humane reasons, but this was not related to the test substance.
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Group mean food consumption for males receiving phytosterol ester at dietary levels of 1.6%, 3.2% and 8.1% was slightly higher than that of concurrent controls on occasions during the study. This was probably due to the lower energy content of the diets due to the phytosterol component and wasn't considered to be of any toxicological significance.
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Description (incidence and severity):
Only minor statistically significant changes observed, which were not related to the test substance.
Slightly reduced platelet counts were observed for females at all doses, in comparison with concurrent controls. However, the magnitude of these changes was not sufficient to be considered biologically significant; for example, a 50-fold increase in dietary phytosterol levels from 0.16% to 8.1%, resulted in only a 3% reduction in platelet count. While slightly reduced platelet counts were also observed for males receiving phytosterol ester at dietary levels of 1.6% and
3.2%, group mean platelet count for males receiving phytosterol ester at the highest dietary level of 8.10% was not statistically different from that of controls.

Marginally reduced prothrombin time as observed in females at all doses and marginally increased APTT time was observed for males receiving phytosterol ester at dietary levels of 3.2% and 8.1%. These differences from control values were very small and were considered to be of no biological or toxicological significance.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Only minor statistically significant changes observed, which were not related to the test substance.

Minor increases in plasma albumin, phosphorus, or magnesium levels and slight increases in several plasma or serum enzymes (including plasma alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatinine kinase and lactate dehydrogenase activities and serum 5'-nucleotidase activity) were observed for males and/or females at dietary phytosterol ester levels of 1.6%, 3.2% and 8.1%. However, in the absence of any significant organ weight changes
or histopathological findings in these animals, the changes were concluded not to be biologically significant.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
There were no statistically significant changes to organ weights in comparison with the control group.
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no macroscopic findings that could be attributed to treatment.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
There were no microscopic findings that could be attributed to treatment.

A slightly increased incidence of microlithiasis was observed in the kidney cortex of females of the high dose group. This is a common finding of variable incidence in the kidneys of female rats of this strain and was not considered to be an adverse effect.
Histopathological findings: neoplastic:
not examined
Dose descriptor:
NOAEL
Effect level:
>= 8.1 other: %
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Dose descriptor:
NOAEL
Effect level:
>= 6 600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
other: phytosterol ester
Dose descriptor:
NOAEL
Effect level:
>= 4 100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
other: phytosterol
Key result
Critical effects observed:
no
Conclusions:
In a 90-day repeated dose toxicity study (reliability score 1) conducted using a protocol equivalent to OECD 408 and in compliance with GLP, administration of phytosterol in the diet of Wistar rats for 90 days did not lead to any adverse effects up to dietary concentrations of 8.1% phytosterol ester (equivalent to 6600 mg/kg bw/day; or 4100 mg/kg bw/day phytosterol).
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-07-28 to 2016-12-02
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
Deviations had no adverse impact on the scientific purpose of the study.
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Remarks:
Han™:RccHan™:WIST strain
Details on species / strain selection:
The rat was selected for this study as it is a readily available rodent species historically used in safety evaluation studies and is acceptable to appropriate regulatory authorities.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited (Oxon, UK)
- Females (if applicable) nulliparous and non-pregnant: not specified
- Age at study initiation: 6-8 weeks
- Weight at study initiation: Males: 167 to 208g; Females: 141 to 170g
- Fasting period before study: Not specified
- Housing: groups of up to four by sex in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding (Datesand Ltd., Cheshire, UK).
- Diet (e.g. ad libitum): Ground diet (Rat and Mouse SQC Ground Diet No. 1, Special Diet Services, Dietex International Limited, Witham, Essex, UK) was used ad libitum
- Water (e.g. ad libitum): Mains drinking water was supplied ad libitum from polycarbonate bottles attached to the cage
- Acclimation period: 8 days

DETAILS OF FOOD AND WATER QUALITY: The diet, drinking water, bedding and environmental enrichment was considered not to contain any contaminant at a level that might have affected the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least fifteen air changes per hour
- Photoperiod (hrs dark / hrs light): low intensity fluorescent lighting was controlled to give twelve hours continuous light and twelve hours darkness

IN-LIFE DATES: From: 2015-08-21 To: 2015-11-20
Route of administration:
oral: feed
Details on route of administration:
The oral route was selected as the most appropriate route of exposure, based on the physical properties of the test item, and the results of the study are considered to be relevant for the evaluation of the toxicological properties of the test item.
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): Dietary admixtures were prepared fortnightly and stored at room temperature
- Mixing appropriate amounts with (Type of food): A known amount of test item was mixed with a small amount of basal laboratory diet in a Robot Coupe Blixer 4 mixer until homogeneous. This pre-mix was then added to a larger amount of basal laboratory diet and mixed for a further sixty minutes at a constant speed, setting 1 in a Hobart H800 mixer.
- Storage temperature of food: stable for 20 days at room temperature
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Representative samples of dietary admixtures were taken and analyzed for concentration of Rosin CAS 8050-09-7 at Envigo Research Limited, Shardlow, UK, Analytical Services. The results indicate that the prepared formulations were within acceptable limits of the nominal concentration, thus confirming the suitability and accuracy of the formulation procedure.
Duration of treatment / exposure:
ninety consecutive days
Frequency of treatment:
continuously in the diet, for ninety consecutive days
Dose / conc.:
0 ppm
Remarks:
control
Dose / conc.:
2 500 ppm
Remarks:
Low concentration
Dose / conc.:
5 000 ppm
Remarks:
Intermediate concentration
Dose / conc.:
7 500 ppm
Remarks:
High concentration
No. of animals per sex per dose:
10/sex/concentration
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: The dietary concentrations were chosen based on the results of a previous toxicity study (Harlan Laboratories Ltd., Study Number: D80926: Tall Oil Rosin - CAS no. 8050-09-7: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test in the Han Wistar Rat).
- Rationale for animal assignment (if not random): The animals were randomly allocated to treatment groups using a stratified body weight randomization procedure and the group mean body weights were then determined to ensure similarity between the treatment groups. The cages were distributed in dose group columns within the holding rack to minimize the potential for cross contamination of the treated diet. The animals were uniquely identified within the study by an ear punching system routinely used in these laboratories.
Positive control:
Not used
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: All animals were examined for overt signs of toxicity, ill-health or behavioural change daily from the start of treatment.

BODY WEIGHT: Yes
- Time schedule for examinations: Individual body weights were recorded on Day 1 (prior the start of treatment) and at weekly intervals thereafter. Body weights were also recorded at terminal kill.

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
Food consumption was recorded for each cage group at weekly intervals throughout the study

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: Not specified

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water intake was observed daily, for each cage group, by visual inspection of the water bottles for any overt changes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: pre-treatment and before termination of treatment (during Week 12).
- Dose groups that were examined: all control and high dose animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Day 90
- Anaesthetic used for blood collection: Not specified
- Animals fasted: No
- How many animals: all animals from each test and control group
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Day 90
- Animals fasted: No
- How many animals: all animals from each test and control group
- Parameters checked in table [No.1] were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Prior to the start of treatment and at weekly intervals thereafter, all animals were observed for signs of functional/behavioural toxicity
- Dose groups that were examined: all animals
- Battery of functions tested: sensory activity / grip strength / motor activity
- Parameters checked included: grasp response, vocalisation, toe pinch, tail pinch, finger approach, touch escape, pupil reflex, blink reflex, startle reflex

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table 2)

HISTOPATHOLOGY: Yes (see table 2)
Statistics:
See "any other information on materials and methods incl. tables" for information on statistics.
Clinical signs:
no effects observed
Description (incidence and severity):
No clinical signs were apparent during the study in animals exposed to diets containing 2500, 5000 or 7500 ppm of the test item.
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Animals of either sex exposed to 7500 ppm showed a statistically significant reduction in body weight gain during the first week of treatment. Recovery was evident in males thereafter, however a statistically significant reduction in body weight gain was evident in these males during Week 11 and overall weight gain was reduced by 6%. Females continued to show reduced body weight gain during Weeks 2, 3, 4, 6 and 10, with statistical significance being achieved during Weeks 4 and 6. Overall body weight gain for 7500 ppm group females was statistically significantly lower (29%) compared with controls. Statistically significantly lower mean body weight compared to controls was apparent in 7500 ppm group females from Day 15 until termination.

Overall body weight gain for males exposed to 5000 ppm and females exposed to 2500 and 5000 ppm was slightly lower than the control group and occasional statistically significant reductions in body weight (5000 ppm females only) and body weight gain for these animals relative to control were also noted. However, these differences did not show an exposure related response and were therefore considered to be unrelated to test item exposure.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
There was no adverse effect of dietary exposure to 2500, 5000 or 7500 ppm of the test item on food consumption for either sex or food conversion efficiency for males.

Reduced food conversion efficiency was evident in females exposed to 7500 ppm and followed the fluctuations seen in body weight gains for these females.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
There was no observed effect of dietary exposure to 2500, 5000 or 7500 ppm of the test item on water consumption for either sex.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
Ophthalmic examination of the eyes from rats receiving diet containing 7500 ppm did not indicate any effect of treatment.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Animals of either sex fed diet containing 7500 ppm showed a statistically significant reduction in haemoglobin and haematocrit. Females exposed to 7500 ppm also showed a statistically significant reduction in erythrocyte count.

Females exposed to 5000 ppm showed a statistically significant reduction in hematocrit and haemoglobin. All of the individual values were within historical control ranges for rats of the strain and age used. In the absence of any associated histopathology correlates in the bone marrow or spleen, the intergroup differences were not considered to be of toxicological importance.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
There were no toxicologically significant effects detected in the blood chemical parameters examined.

Males from all dietary groups showed a statistically significant increase in bilirubin and a statistically significant reduction in bile acids. Females from all dietary groups showed a statistically significant increase in urea and a statistically significant reduction in alanine aminotransferase. Males fed diets containing 7500 ppm showed a statistically significant increase in cholesterol. Females fed diet containing 7500 ppm showed statistically significant reductions in glucose, total protein, albumin and bile acids and statistically significant increases in bilirubin and chloride concentration. The effect on total protein, albumin, bilirubin and bile acids extended to females fed diet containing 5000 ppm. Females fed diet containing 5000 ppm also showed a statistically significant increase in creatinine and a statistically significant reduction in phosphorus. Males fed diet containing 5000 ppm showed a statistically significant increase in cholesterol and a statistically significant reduction in albumin. Females fed diet containing 2500 ppm showed a statistically significant increase in creatinine. Individual values were within the historical control ranges and in the absence of exposure related responses (excluding bilirubin for females) or any associated histopathological correlates, the intergroup differences were not considered to be toxicologically significant.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
Behavioural Assessments
Weekly assessment of the animals in a standard arena did not reveal any obvious adverse effects of dietary exposure to 2500, 5000 or 7500 ppm of the test item.

Functional Performance Tests
There were no toxicologically significant effects detected in functional performance.

Females exposed to 7500 ppm showed a statistically significant increase in hindlimb grip strength (two out of the three tests, p<0.05 or p<0.01). Females exposed to 7500 and 2500 ppm also showed a statistically significant increase in forelimb grip strength (one out of the three tests p<0.01). There was generally no dose-relationship and in the absence of any supporting clinical observations or pathological change to suggest a neurotoxic effect, the intergroup differences were considered to be of no toxicological significance.

Sensory Reactivity Assessments
There were no differences observed in the scores for sensory reactivity for either sex during the study.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Females exposed to diets containing 7500 and 5000 ppm showed a statistically significant and exposure related reduction in absolute adrenal weights. However, although relative (to terminal body weight) adrenal weights of females in the 7500 and 5000 ppm groups were statistically significantly lower than control, they did not follow a treatment related response. The majority of individual values for both absolute and relative weights for treated females were within historical control ranges, whereasindividual values for two control females exceeded the historical absolute ranges and three control females exceeded the historical control range for relative adrenal weight.

There was considered to be no effect of dietary exposure in 7500 or 5000 ppm males or in 2500 ppm animals of either sex on the organ weights measured.

In males fed diets containing 7500 ppm absolute and relative liver, brain and kidney weights were statistically significantly increased relative to control. The majority of the individual values were within the historical control range and in the absence of any histopathological correlates these intergroup differences were considered of no toxicological significance. Females fed diet containing 7500 and 5000 ppm showed a statistically significant reduction in absolute and relative (to terminal body weight) uterus/cervix weight but without any relationship to exposure. The reduction in the weight in the uterus/cervix in 7500 ppm group females was considered to be due to the stage of the reproductive cycle (fewer animals in proestrus/estrus) and of no toxicological significance.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Neither the type, incidence nor distribution of findings observed at terminal necropsy indicated any obvious effect of dietary exposure to 2500, 5000 or 7500 ppm of the test item. Incidences of reddened lungs were noted in a number of control and test item exposed
animals at necropsy and small testes and epididymides were evident in one male in the 5000 ppm group. These were all considered to be incidental findings.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
In the adrenals, mild hypertrophy of the zona glomerulosa was present in four males and four females fed diets containing 7500 ppm of the test item, and in two males and one female fed diets containing 5000 ppm. Hypertrophy of the zona glomerulosa is generally considered to be an adaptive process, usually linked to maintenance of electrolytes and fluid balance (Domenici Lombardo, 1990 and Greaves 2007). In this study, taking the blood chemistry values into account, changes in the zona glomerulosa of the adrenal gland were not considered to have affected normal electrolyteregulation. The histopathological changes in the adrenal glands at 7500 and 5000 ppm did not appear to have an effect on homeostatic control, and therefore were not considered to have had adverse consequences for the organ or the body. No similar effects were detected in animals of either sex fed diet containing 2500 ppm of the test item.

No other findings were present at histopathology which correlated with in-life changes noted. In particular the apparent reduction in weightin the uterus/cervix in high dose females was considered likely to be due to the stage of the reproductive cycle (fewer animals in proestrus/estrus) and of no toxicological significance.
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
5 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
Critical effects observed:
no

Table 7. Group Mean Body Weight Gains – Male Rats

 

 

 

Group

 

Increase in Body Weight (g)

 

Day Numbers Relative to Start Date

Abs Gain

% Gain

From

1

8

15

22

29

36

43

50

57

64

71

78

85

1

1

To

8

15

22

29

36

43

50

57

64

71

78

85

91

91

91

 

Control

(0 ppm)

Mean

34.9

23.8

23.6

16.8

13.7

15.3

10.5

10.5

8.4

4.5

12.3

2.3

7.0

183.6

99.3

S.D.

7.5

3.9

5.0

5.5

4.9

7.5

2.8

5.7

4.5

3.0

4.4

3.5

3.1

39.7

21.5

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

 

Low Concn

(2500 ppm)

Mean

37.3

20.6

22.3

17.1

16.9

16.1

13.1

12.4

9.9

6.8

11.1

5.4

4.0

193.0

103.5

S.D.

3.5

3.8

6.5

4.5

9.3

9.7

3.9

3.6

4.1

2.6

4.7

3.5

4.7

28.9

16.6

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

 

Intermediate

Concn

(5000 ppm)

Mean

30.9

20.3

18.2

11.0*

14.0

13.7

11.6

13.7

9.5

6.9

7.5*

6.6*

5.3

169.2

91.6

S.D.

5.7

5.2

5.5

5.6

3.6

3.6

4.6

2.1

3.1

3.4

4.5

2.3

4.3

25.9

15.1

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

 

High Concn

(7500 ppm)

Mean

27.9**

21.5

18.1

13.2

17.7*

17.3

12.5

13.9

8.3

5.7

7.3*

3.8

5.8

173.0

95.0

S.D.

4.6

5.9

7.1

5.6

19.5

18.0

3.7

4.6

4.7

2.7

4.3

3.9

3.0

26.0

15.8

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 8. Group Mean Body Weight Gains – Female Rats

 

 

 

Group

 

Increase in Body Weight (g)

 

Day Numbers Relative to Start Date

Abs Gain

% Gain

From

1

8

15

22

29

36

43

50

57

64

71

78

85

1

1

To

8

15

22

29

36

43

50

57

64

71

78

85

91

91

91

 

Control

(0 ppm)

Mean

14.3

14.5

9.2

7.7

4.5

7.4

3.9

5.2

2.2

4.1

3.1

0.5

3.1

79.7

51.0

S.D.

5.8

1.9

3.7

3.0

3.3

2.8

2.3

2.8

4.0

3.9

5.0

2.8

4.5

15.3

8.8

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

 

Low Concn

(2500 ppm)

Mean

11.9

12.3

10.4

5.8*

7.4

2.8*

4.1

4.8

2.6

2.1

4.6

0.6

-0.7

68.7

44.3

S.D.

3.9

3.6

3.4

2.0

3.4

4.5

2.7

2.6

3.9

2.1

2.9

2.0

3.3

9.8

7.3

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

 

Intermediate

Concn

(5000 ppm)

Mean

11.8

13.1

8.6

4.8*

6.4

6.0*

3.7

4.8

4.2

1.6

-1.5*

1.8

0.5

65.8*

43.7

S.D.

3.9

2.6

3.7

2.7

3.2

2.2

4.5

2.1

3.0

3.5

3.0

2.1

4.0

7.6

5.7

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

 

High Concn

(7500 ppm)

Mean

7.2**

11.8

7.0

5.3*

5.9

2.5**

3.7

5.5

2.1

1.7

2.4

0.9

0.5

56.5***

37.2

S.D.

7.8

6.1

4.8

2.1

3.3

1.6

3.5

3.6

2.3

2.6

2.7

3.4

4.5

10.5

7.3

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 9. Group Mean Body Weight Values – Female Rats

Group

Body weights (g)

Day Numbers Relative to Start Date

From

1

8

15

22

29

36

43

50

57

64

71

78

85

91

Control

(0 ppm)

Mean

155.7

170.1

184.6

193.8

201.5

206.0

213.4

217.3

222.5

224.7

228.8

231.9

232.4

235.5

S.D.

7.0

10.3

10.2

11.7

13.8

16.4

15.3

15.1

16.2

18.9

19.9

16.9

18.0

19.9

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Low Concn

(2500 ppm)

Mean

155.9

167.8

180.1

190.5

196.3

203.7

206.5

210.6

215.4

218.0

220.1

224.7

225.3

224.6

S.D.

9.0

10.0

8.8

7.6

7.6

7.9

9.3

8.0

9.2

11.5

11.8

11.4

11.3

12.0

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Intermediate

Concn

(5000 ppm)

Mean

151.2

163.0

176.1

184.7

189.5

195.9

201.9

205.6

210.4

214.6

216.2

214.7*

216.5

217.0*

S.D.

8.0

7.0

8.1

11.3

10.4

9.8

9.4

12.2

12.0

10.3

12.1

12.0

11.7

10.3

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

High Concn

(7500 ppm)

Mean

152.1

159.3

171.1

*

178.1

*

183.4

**

189.3*

191.8

**

195.5

**

201.0

**

203.1

**

204.8

**

207.2

**

208.1

**

208.6

**

S.D.

9.0

12.2

13.5

17.0

16.5

16.2

15.9

16.3

15.6

15.3

16.2

15.6

16.2

13.9

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Conclusions:
The continuous dietary administration of the test material to rats, at dietary concentrations of 2500, 5000 and 7500 ppm for ninety consecutive days, resulted in reduced body weight gains in animals of either sex exposed to 7500 ppm and microscopic adrenal changes in animals of either sex exposed to 7500 and 5000 ppm. The No Observed Effect Level (NOEL) for both sexes was therefore considered to be 2500 ppm (equivalent to a mean achieved dosage of 174.1 mg/kg bw/day for males and 196.4 mg/kg bw/day for females).

The microscopic adrenal changes (hypertrophy of the zona glomerulosa) identified in animals of either sex exposed to 7500 or 5000 ppm were mild. The changes were not considered to have affected normal electrolyte regulation and did not appear to have an effect on homeostatic control. Therefore, they were not considered to have had adverse consequences for the organ or the body. For this reason, 5000 ppm (equivalent to a mean achieved dosage
of 335.2 mg/kg bw/day for males and 401.2 mg/kg bw/day for females) was considered to be the “No Observed Adverse Effect Level” (NOAEL) systemic toxicity.
Executive summary:

In a key oral repeat dose toxicity study, the test material (Rosin, CAS# 8050-09-7) was administered continuously in the diet of three groups, each composed of ten male and ten female Wistar Han™:RccHan™:WIST strain rats, for ninety consecutive days, at dietary concentrations of 2500, 5000 or 7500 ppm (equivalent to a mean achieved dosage of 174.1, 335.2 or 510.1 mg/kg bw/day for males and 196.4, 401.2 or 596.2 mg/kg bw/day for females). A control group of ten males and ten females were fed basal laboratory diet.

 

Clinical signs, functional observations, body weight change, dietary intake and water consumption were monitored during the study. Haematology and blood chemistry were evaluated for all animals at the end of the study. Ophthalmoscopic examination was also performed on control group and high dose animals before the start of treatment and during Week 12 of the study. All animals were subjected to gross necropsy examination and a comprehensive

histopathological evaluation of tissues was performed.

 

No mortality was observed through the study period. There were no clinical signs observed that indicated any adverse effects of dietary exposures

at 2500, 5000 or 7500 ppm. Behavioural, functional, and sensory reactivity parameters remained unaffected by exposure to the test material and there were no adverse treatment-related effects on the eyes, water consumption, hematological, or clinical chemistry parameters evaluated. Gross necropsy did not reveal any remarkable findings.

 

Animals of either sex exposed to diets containing 7500 ppm showed a reduction in body weight gain during the first week of treatment. Recovery was evident in males thereafter, however a reduction in body weight gain was evident in these males during Week 11 and overall body weight gain in males was reduced relative to controls. Females continued to show a reduced body weight gain during Weeks 2, 3, 4, 6 and 10. Overall body weight gain for these females was reduced when compared to controls. Reduced food conversion efficiency was evident in females exposed to diets containing 7500 ppm, and followed the fluctuations seen in body weight gains for these females. No toxicologically significant effects on body weight were detected in animals of either sex exposed to diets containing 5000 or 2500 ppm.

 

Females exposed to diets containing 7500 and 5000 ppm showed a statistically significant reduction in adrenal weights, both absolute and relative to terminal body weight. However, relative adrenal weights of females in the 7500 and 5000 ppm groups did not follow an exposure related response and a majority of individual values for both absolute and relative weights for treated females were within historical control ranges. There was considered to be no effect of dietary exposure in 7500 or 5000 ppm males or in 2500 ppm animals of either sex on the organ weights measured.

 

Histopathological evaluation revealed hypertrophy of the zona glomerulosa in four males and four females exposed to 7500 ppm and in two males and one female exposed to 5000 ppm (all at a mild level). No such effects were detected in animals of either sex exposed to 2500 ppm. Hypertrophy of the zona glomerulosa is generally considered to be an adaptive or reactive change rather than an adverse effect of treatment (Domenici Lombardo, 1990 and Greaves, 2007) and was therefore not considered to be treatment-related.

 

The continuous dietary administration of the test material to rats, at dietary concentrations of 2500, 5000 and 7500 ppm for ninety consecutive days, resulted in reduced body weight gains in animals of either sex exposed to 7500 ppm and microscopic adrenal changes in animals of either sex exposed to 7500 and 5000 ppm. The No Observed Effect Level (NOEL) for both sexes was therefore considered to be 2500 ppm (equivalent to a mean achieved dosage of 174.1 mg/kg bw/day for males and 196.4 mg/kg bw/day for females).

 

The microscopic adrenal changes (hypertrophy of the zona glomerulosa) identified in animals of either sex exposed to 7500 or 5000 ppm were mild. The changes were not considered to have affected normal electrolyte regulation and did not appear to have an effect on homeostatic control. Therefore, they were not considered to have had adverse consequences for the organ or the body. For this reason, 5000 ppm (equivalent to a mean achieved dosage

of 335.2 mg/kg bw/day for males and 401.2 mg/kg bw/day for females) was considered to be the “No Observed Adverse Effect Level” (NOAEL) systemic toxicity.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
335.2 mg/kg bw/day
Study duration:
subchronic
Species:
rat
System:
other: reduced body weight gain

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 study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Tall Oil Soap is a complex UVCB substance containing a range of constituent types. The principal constituents are sodium salts of saturated and unsaturated C14-C20 fatty acids (5-45% w/w, typical ca. 26%), rosin acid sodium salts (10-40% w/w, typical ca. 23%) and sterols (1-10%, typical ca. 3.5%). Other neutral constituents such as rosin alcohol and aldehyde isomers are also present at 0 – 3% w/w (typically ca. 1.3%).

In order to reduce in vivo testing in vertebrates, the REACH Regulation requires registrants to consider alternative methods to fulfil Annex requirements. For endpoints where measured data for substance are not available, a weight of evidence approach has therefore been followed based on available data for groups of constituents.

The registrants of TOS consider that the available information for constituents of the substance are sufficient for the purposes of hazard assessment and risk characterisation and that further animal testing is not warranted.

Reliable repeated dose toxicity data and other supporting information are available for substances (including UVCBs) that are representative of these four main constituent groups, correlating to Blocks, 1, 2, 9 and 11 in the substance definition (Section 1.2) and environmental assessment.

In addition, TOS contains 25-45% (typical ca. 32%) w/w/ water and cellulose fibre, lignin and oligomeric acids at 3 – 30% w/w (typical ca. 10%). These are not considered further for the assessment of human health hazards.

Other minor constituent groups, for which relevant measured data have not been identified, are typically present at concentrations that are below the threshold to be taken into consideration in the rules for classification and labelling of mixtures for specific target organ toxicity according to Regulation (EC) No 1272/2008 (Table 3.9.4). These constituent groups are therefore not assessed separately since the conditions of use specified for the whole substance or Blocks 1, 2 and 11 would be sufficient to control any risk relating to hazards that have not been identified for the remaining blocks.

Block 1: Fatty acids

Tall Oil Fatty Acids (TOFA, CAS 61790-12-3) is exempt from registration under REACH in accordance with Annex V of the Regulation. It is non-hazardous product containing fatty acids in the range C14 to C24. As such, the fatty acid constituents of CTO and TOS can also be considered non-hazardous. There are no readily available, reliable repeated dose toxicity data for TOFA.

On this basis, the hazard conclusion for long-term systemic toxicity of TOS Block 1 is ‘no hazard identified’.

Block 2: Rosin acids and Block 9: Rosin alcohol and aldehyde isomers

A 90-day oral repeated dose toxicity test (Envigo, 2017) conducted to OECD 408 and in compliance with GLP is available for tall oil rosin, which represents the neutral form of the rosin acid sodium salts present at 10-40% by weight in the registration substance. In this study, rosin (CAS 8050-09-7) was administered continuously in the diet of male and female Wistar rats for 90 days at concentrations of 2500, 5000 or 7500 ppm (equivalent to mean achieved doses of 174.1, 335.2 or 510.1 mg/kg bw/day for males and 196.4, 401.2 or 596.2 mg/kg bw/day for females). Reduced body weight gain was noted in animals of either sex in the highest dose group of 7500 ppm and microscopic adrenal changes in animals of either sex exposed to 7500 and 5000 ppm. The microscopic adrenal changes (hypertrophy of the zona glomerulosa) were not considered to have affected normal electrolyte regulation and did not appear to have an effect on homeostatic control. These changes were not considered to have had adverse consequences for the organ or the body. Therefore, the NOAEL was concluded to be 5000 ppm (equivalent to a mean achieved dose of 335.2 mg/kg bw/day for males and 401.2 mg/kg bw/day for females) based on reduced body weight gain in the highest dose group. No other adverse effects were observed.

The main constituents of the test material were pimaric acid (CAS 127-27-5), palustric acid (CAS 1945-53-5), isopimaric acid (CAS 5835-26-7), abietic acid (CAS 514-10-3) and neoabietic acid (CAS 471-77-2), which are representative of the sodium salts of rosin acid constituents (Block 2) of the registration substance (see section 1.2 for details).

Tall oil rosin also typically contains small amounts of neutral constituents that represent Block 9, rosin alcohol and aldehyde isomers. No specific information is available with regard to the concentration of these constituents present in the test material used for the 90-day repeated dose toxicity study. However, in the absence of toxicological  data for any known substances in this block, and the low concentrations of any individual constituent in TOS, the result for tall oil rosin is used as a surrogate for quantitative risk characterisation of Block 9.

Based on these data, it can be concluded that no classification for specific target organ toxicity following repeated exposures is required. However, the observed effects on body weight gain are taken into consideration for the overall hazard conclusion, and a subchronic oral NOAEL of 335.2 mg/kg bw is taken as the starting point for determining quantitative DNELs for Blocks 2 and 9.

Block 11: Sterols

The sterol constituents of TOS are also present in many other plant species, including foodstuffs. In particular β-sitosterol is added at significant levels to foods due to its cholesterol-lowering properties. Plant sterols have been studied in repeated dose and reproductive toxicity tests.

In a 90-day repeated dose toxicity study (Hepburn et al., 1998) conducted using a protocol equivalent to OECD 408 and in compliance with GLP, administration of phytosterol continuously in the diet of male and female Wistar rats for 90 days did not lead to any adverse effects at dietary concentrations of up to 8.1% phytosterol ester (equivalent to 6600 mg/kg bw/day; or 4100 mg/kg bw/day phytosterol).

Similarly, no general adverse effects were observed in parent generations in a dietary reproductive toxicity study on phytosterol esters (see Section 7.8), conducted using a protocol comparable with OECD 416 and in compliance with GLP (Waalkens-Berendsen et al., 1999). The NOAEL from this study was 2500-9100 mg/kg bw/day phytosterol ester or 1540-5620 mg/kg bw/day phytosterol.

These good quality studies used sterol/fatty acid esters as test material, since this is the chemical form in which they are extracted from plants. TOS itself contains mainly sodium salts of the fatty acids and “free” sterol. However, this does not negate use of the data to support REACH registrations of CTO-derived materials because in vivo, the esters are hydrolysed by pancreatic carboxy ester lipase, therefore the toxicological properties of the free sterols are of most relevance (EC, 2000).

The composition of the tested material was as shown in Tables 5.6.2 and 5.6.3 below.

Table 5.6.2: Gross composition of the test material

Constituent

Composition (% w/w)

Total sterol

62.0

Total fatty acid

38.2

Free sterol (as % of the total mixture)

8.4

Free fatty acid

<0.3

Table 5.6.3: Sterol profile of the test material

Sterol Constituent

% w/w

Fatty acid profile

% w/w

Cholesterol

0.4

C16:0

9.6

Brassicasterol

1.1

C18:0

4.1

Campesterol

25.8

C18:1

21.6

Stigmasterol

21.6

C18:2

64.6

β-Sitosterol

48.7

 

 

β-Sitostanol

1.8

 

 

Unknowns

0.8

 

 

Reference: EC, 2000. European Commission Health and Consumer Protection Directorate-General, Opinion of the Scientific Committee on Food on a Request for a Safety Assessment of the use of Phytosterol Esters in Yellow Fat Spreads.

On the basis of these data, the hazard conclusion for long-term systemic toxicity of TOS Block 11 is ‘no hazard identified'.

Distilled tall oil (CAS 8002-26-4, EC No. 232-304-6)

In addition to the data described above, subacute toxicity data are available for the related substance Distilled Tall Oil (DTO). DTO is obtained by physical distillation of CTO and the two substances therefore share many of the same constituents in different proportions. Direct read-across from DTO to CTO and TOS is not appropriate because DTO does not contain any sterol constituents and has lower neutral content overall. However, an existing OECD 422 combined repeated dose toxicity test and reproductive/developmental screening test adds weight of evidence to the absence of adverse effects following repeated exposure to CTO and TOS.

In the Combined Repeated Dose Toxicity Study with Reproduction/Developmental Toxicity Screening test (Inveresk, 2002), conducted according to OECD 422 and in compliance with GLP, four groups of 10 male and 10 female Sprague-Dawley rats received tall oil via the diet at concentrations of 0, 1000, 5000 and 20000 ppm. The males were dosed for at least four weeks, starting from two weeks prior to mating. The females were dosed from two weeks prior to mating until at least day 6 of lactation. The animals were monitored for clinical signs, body weight, food consumption, mating and litter performance. Blood samples were taken from five males and five females per group for haematology and clinical chemistry investigations. Necropsy was conducted on all animals, which included weighing of major organs.  Histopathology was conducted on tissues from five males from control and high dose groups, and seven females from the control and eight females from the high dose groups.

At 20000 ppm in-life observations included decreased weight gain and food consumption in both sexes. Increased male liver weight following covariance analysis and increases in bilirubin and alkaline phosphatase were noted in both sexes. At 5000 ppm liver weight in males and alkaline phosphatase in both sexes were increased. In conclusion, under the conditions of this study, the parental NOEL was reported to be 1000 ppm. However, the effects on the liver are considered (by the author the study summary) to be adaptive rather than adverse, and decreased food consumption and weight gain were most likely to be due to palatability of the test diet, rather than toxicity. Therefore, the overall NOAEL for general toxicity was 5000 ppm tall oil (equivalent to approximately 423 mg/kg bw/day for males and 483 mg/kg bw/day for females; the highest dose tested).

Conclusions for repeated dose toxicity of TOS

Subchronic and subacute data on the constituents of TOS (rosin acids and neutrals, and sterols) together with supporting subacute data on DTO, lead to the conclusion that TOS does not require classification for specific organ toxicity following repeated oral exposures. The most conservative NOAELs from the available studies on constituents of TOS and DTO is 335.2 to 401.2 mg/kg bw/day (for males and females, respectively) based on reduced body weight gain in the subchronic toxicity study with tall oil rosin. Since the constituents of tall oil rosin represent only around 43% by weight of TOS at most, and no hazards have been identified for the other constituent groups, the true NOAEL of the whole substance is likely to be higher. However, as a conservative approach, the lowest available subchronic NOAEL of 335.2 mg/kg bw as a starting point for the overall systemic hazard conclusions for TOS.

Justification for classification or non-classification

Based on data for constituents of TOS, no classification for specific target organ toxicity for repeated exposures is required in accordance with Regulation (EC) No 1272/2008.