Registration Dossier

Administrative data

Workers - Hazard via inhalation route

Systemic effects

Long term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
Acute/short term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
DNEL (Derived No Effect Level)
Value:
10 mg/m³
DNEL related information
DNEL derivation method:
other: ECHA Guidance. Generic ECHA recommendation for a long-term DNEL (inhalation, worker)
Acute/short term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
DNEL related information

Workers - Hazard via dermal route

Systemic effects

Long term exposure
Hazard assessment conclusion:
DNEL (Derived No Effect Level)
Value:
2.09 mg/kg bw/day
Most sensitive endpoint:
repeated dose toxicity
Route of original study:
Oral
DNEL related information
DNEL derivation method:
ECHA REACH Guidance
Overall assessment factor (AF):
100
Dose descriptor starting point:
NOAEL
Value:
209.1 mg/kg bw/day
Modified dose descriptor starting point:
NOAEL
Value:
209.1 mg/kg bw/day
Explanation for the modification of the dose descriptor starting point:

Modification of dose descriptor:

 

Converted oral NOAEL rat (in mg/kg bw/day) into dermal NOAEL rat (in mg/kg bw/day) by correcting for differences in absorption between routes as well as for differences in dermal absorption between rats and humans: 

 

corrected dermal NOAEL = oral NOAEL x (ABSoral-rat / ABSderm-rat) x (ABSderm-rat / ABSderm-human)

 

                                      = oral NOAEL x (ABSoral-rat / ABSderm-human)

 

                                       = 209.1 mg/Kg bw/day x (1 / 1) 

 

Note: Dermal absorption assumed not be higher than oral absorption, therefore no default factor (i.e. factor 1) introduced when performing oral-to-dermal extrapolation (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).

AF for dose response relationship:
1
Justification:
Default assessment factor when the starting point for the DNEL calculation is a NOAEL (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for differences in duration of exposure:
2
Justification:
Default assessment factor of 2 applied when extrapolating duration of exposure from sub-chronic to chronic (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for interspecies differences (allometric scaling):
4
Justification:
Allometric scaling factor for rats compared to humans (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for other interspecies differences:
2.5
Justification:
Additional factor of 2.5 for other interspecies differences; systemic effects (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for intraspecies differences:
5
Justification:
For workers, as standard procedure for threshold effects, a default assessment factor of 5 was applied (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for the quality of the whole database:
1
Justification:
Default assessment factor applied for good/standard quality of the database, taking into account completeness, consistency and the standard information requirements (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified

Workers - Hazard for the eyes

Local effects

Hazard assessment conclusion:
medium hazard (no threshold derived)

Additional information - workers

Acute toxicity

ECHA Guidance R.8 (Chapter R.8.1.2.5) indicates that DNELs for acute toxicity are not required if no acute toxicity hazard leading to classification has been identified. The members of this category are not acutely toxic following oral or dermal exposure (LD50 >2000 mg/Kg bw) while a low vapour pressure precludes inhalation exposure indicating a low of concern for this route of exposure. No DNELs for acute toxicity are therefore necessary.

 

Irritation

Corrosive and irritant effects on the skin and eye are local, concentration-dependent phenomena. However, while test results indicate that members of this category are irritating to eye (but not to skin), the nature of the data is such that no conclusion can be drawn with regard to any

dose-response relationship present. No DNEL for irritation can therefore be derived.

 

Sensitisation

The sensitisation potential of the substances included in this category is well understood and comprises results from six local lymph node assays and a guinea pig maximisation test. The LLNA results consistently showed evidence of a potential to induce skin sensitisation.

Intrinsic sensitising potency

ECHA Guidance R.8, Appendix R.8-10, (ECHA, 2010) states that while skin sensitisation is generally regarded as a threshold effect it may be very difficult to derive a threshold and to set a DNEL. Thus, the general approach for sensitisers involves a qualitative approach where a DNEL is used to judge any remaining/residual risks after the implementation of appropriate risk management measures (RMM) and occupational controls (OC). The extent of the RMM and OC required is dependent on the intrinsic sensitising potency of the substance.

 

For results obtained using the LLNA, intrinsic sensitising potency is based on the EC3 and defined (ECHA (2010), Appendix R.8-10) as follows:

 

Category

EC3 (%)

Extreme

<0.2%

Strong

>0.2 - <2

Moderate

>2

 

An EC3 of 14% was obtained with Resin acids and rosin acids, fumarated esters with pentaerythritol while three samples of Resin acids and rosin acids, maleated esters with pentaerythritol returned EC3 values of 16%, 17% and 22%. No EC3 could be derived from a LLNA test with a Resin acids and rosin acids, fumarated esters with glycerol and pentaerythritol since stimulation indices in a range 4.37-4.73 (obtained over a concentration range of 10-50% test substance) resulted in a negative intercept when the data were subject to linear regression analysis. Overall, however, these results indicate that members of this category have a moderate potential to cause skin sensitisation.

 

Results obtained from a guinea pig maximisation test using an intradermal induction concentration of 7.5% indicate that Resin acids and rosin acids, fumarated esters with pentaerythritol was not a skin sensitiser. Although some anomalous results were obtained, the information summarised above indicates that the members of this category have a moderate potential to cause sensitisation following skin contact.

 

Derivation of a DNEL for sensitisation

ECHA Guidance R.8, Appendix R.8-10 (ECHA, 2010), indicates that the EC3 concentration from a LLNA test can be taken as a LOAEL for the induction of skin sensitisation (ECHA, 2010) after conversion into an equivalent dose per unit area of skin (µg/cm2). Assuming (i) a dose volume of 25 μL (according to

the standard LLNA protocol); (ii) an estimated treatment area of 1 cm2for the mouse ear; and (iii) an assumed density of is 1, the conversion is performed as follows:

 

EC3 [%] * 250 [µg/cm2/%] = EC3 [µg/cm2]

 

Based on an EC3 of 14% obtained for Resin acids and rosin acids, fumarated esters with pentaerythritol,the equivalent EC3 [µg/cm2] is therefore:

 

14% * 250 = 3500 µg/cm2

 

EC3 [µg/cm2] values of 4000, 4250 and 5500 µg/cm2 were obtained from LLNA data for Resin acids and rosin acids, maleated esters with pentaerythritol.

 

The EC3 of 3500 µg/cm2 obtained for Resin acids and rosin acids, fumarated esters with pentaerythritol will be used to assess the magnitude of any remaining/residual risks after the use of RMMs and OCs recommended in the Qualitative Risk Assessment for this category. No Assessment Factors will be applied to this value since (relative to EC3 values available for other category members) it is already a conservative result, with skin penetration (and hence the capacity of the substance to induce skin sensitisation) enhanced in the LLNA test by the deliberate use of a solvent system that is not present in an occupational or consumer setting. The EC3 of 3500 µg/cm2 will therefore be used as a human NAEL (No Adverse Effect Level).

 

Repeated dose toxicity

 

Information exists to characterise the repeated dose toxicity of Rosin adduct esters. These are formed after modification of rosin with either fumaric acid or maleic anhydride followed by esterification with glycerol and/or pentaerythritol, and hence the adduct ester products therefore exhibit close structural similarities. The available data includes results obtained from testing Resin acids and rosin acids, fumarated, esters with glycerol; Resin acids and rosin acids, fumarated, esters with pentaerythritol; and Resin acids and rosin acids, maleated, esters with pentaerythritol with supporting information on Rosin, fumarated (precursor substance). This information is summarised below.

 

In a key oral repeated dose toxicity test conducted according to OECD Guideline 408 (Envigo Research Limited, 2017a), the test material (Resin acids and Rosin-acids, maleated, esters with pentaerythritol’ CAS# 94581-17-6) was administered in the diet to three groups, each composed of ten male and ten female Wistar Han™:RccHan™:WIST strain rats, for ninety consecutive days, at dietary concentrations of 3000, 6000 or 12000/15000 ppm (equivalent to mean achieved dosages of 209.1, 413.2 or 903.7 mg/Kg bw/day for males and 248.7, 436.7 or 1069.4 mg/Kg bw/day for females). The highest dietary level was increased from 12000 ppm to 15000 ppm after four weeks of treatment to maximise the achieved dosage for this dose group as the study progressed. A control group composed of ten males and ten females was fed basal laboratory diet.

  

No unscheduled mortality was observed through the study period and there were no obvious clinical effects of dietary exposure to test material at concentrations of 3000, 6000 or 12000/15000 ppm for either sex. Behavioural assessment, functional performance, and sensory reactivity assessments remained unaffected post treatment with the test material and there were no effects observed on food conversion efficiency, water consumption, or ophthalmoscopic parameters evaluated in animals of either sex. 

 

Dietary exposure to the test material at a concentration of 12000/15000 ppm was associated with adverse histopathological findings in the liver and urinary bladder. At a dietary exposure level of 6000 ppm, similar histopathological changes were apparent in both the liver and urinary bladder and, although the incidence was lower than observed at 12000/15000 ppm, these findings were considered to be adverse. At the 3000 ppm dietary exposure level, the adverse histopathological changes observed at higher exposure levels were not apparent and histopathological changes were limited to a low incidence of increased adipose tissue in the sternum bone marrow, which was considered to reflect individual variation and to be of no toxicological significance. Although occasional differences from control were observed for some blood chemistry parameters at the 3000 ppm level they were not regarded as adverse effects. Consequently, the No Observed Adverse Effect Level (NOAEL) of RARA, maleated, esters with pentaerythritol when administered via the diet for 90 consecutive days was considered to be 3000 ppm (equivalent to mean achieved dosages of 209.1 mg/kg bw/day for males and 248.7 mg/kg bw/day for females).

 

In a second key oral sub-chronic toxicity study (Envigo Research Limited, 2016a), the test material (Resin acids and Rosin acids, fumarated, esters with pentaerythitol; CAS# 94581-15-4) was administered by continuous dietary exposure to Wistar Han™:RccHan™:WIST strain rats (10/sex/concentration), for ninety consecutive days, at dietary concentrations of 3000 and 6000 ppm for the low and intermediate concentration groups (equivalent to a mean achieved dosage of 210.0 and 414.1 mg/Kg bw/day for males and 262.4 and 511.2 mg/Kg bw/day for females). For the high concentration group, rats were initially fed diet containing 12000 ppm for two weeks followed by 15000 ppm for four weeks and subsequently 18000 ppm for the remainder of the study (equivalent to a mean achieved dosage of 1090.0 mg/Kg bw/day for males and 1298.9 mg/Kg bw/day for females). A control group of ten males and ten females were treated with basal laboratory diet.

  

The continuous oral (dietary) administration of the test material for ninety consecutive days, did not result in any toxicologically significant effects. Therefore, based on the lack of adverse treatment-related effects observed through the study period, the No Observed Adverse Effect Level (NOAEL) for systemic toxicity for both sexes was determined to be 18000 ppm (equivalent to a mean achieved dosage of 1090.0 mg/Kg bw/day for males and 1298.9 mg/Kg bw/day for females).

 

In another key oral sub-chronic toxicity study (Envigo Research Limited, 2015a), the test material (RARA, fumarated, esters with glycerol; CAS# 97489-11-7) was administered by continuous dietary admixture Wistar Han™:RccHan™:WIST strain rats (10/sex/concentration), for ninety consecutive days, at nominal dietary concentrations of 3000, 7500 and 18000 ppm (equivalent to mean achieved dosages of 211.9, 574.0 and 1296.1 mg/Kg bw/day for males and 248.1, 598.9 and 1482.2 mg/Kg bw/day for females). A control group of ten males and ten females were treated with basal laboratory diet.

 

Continuous oral dietary administration of the test material resulted in reduced body weight gain in males treated with 18000 ppm. There were no clinical signs evident for animals of either sex treated with 3000, 7500 or 18000 ppm. The body weight development of high dose males was reduced relative to controls during Weeks 1, 3, 4 and 7 of treatment. Subsequently a 10% reduction in overall body weight gain was evident in these males. No consistent effect on body weight gain was evident in females and no adverse effect was evident in food consumption for either sex. Therefore, the effect on body weight gain in males was most likely associated with test item toxicity rather than a palatability effect.

 

Some statistically significant differences in hematological and blood chemistry parameters were observed in treated animals relative to controls. However, majority of individual values were within the expected normal ranges for rats of the strain and age used. Therefore, these differences were not considered to be toxicologically significant. Macroscopic and microscopic examinations did not reveal any treatment related effects in treated animals.

 

Based on the results of this study, the No Observed Adverse Effect Level (NOAEL) for RARA, fumarated, esters with glycerol was determined to be 18000 ppm for female rats and 7500 ppm for male rats.

 

Resin acids and rosin acids, fumarated, esters with glycerol (CAS# 97489-11-7) was administered to rat by gavage at concentrations of 3000, 7500, or 18000/15000 ppm (Harlan Laboratories Ltd., 2014). The result of the study showed microscopic lung changes in animals of either sex treated with 18000/15000 ppm, microscopic urinary bladder changes in females treated with 18000/15000 ppm and microscopic prostate changes in males 18000 or 7500 ppm. The ‘No Observed Effect Level (NOEL) for systemic toxicity was considered to be 7500 ppm for either sex. 

 

Resin acids and rosin acids, fumarated, esters with pentaerythritol was administered by oral gavage to male and female rats at treatments levels of 0, 30, 300 or 1000 mg/Kg bw/g for approximately 6-7 weeks (Harlan Laboratories Ltd, 2010b). Treatment-related changes in bladder (comprising epithelial hyperplasia, ulceration and erosion with associated subepithelial oedema and inflammation) seen at 1000 mg/Kg/day but not at any other treatment level. The condition was more prevalent among females. A NOAEL of 300 mg/Kg/day for systemic toxicity was established by this study.

 

In a supporting study, Rosin, fumarated (the precursor of Rosin, fumarated, esters with pentaerythritol) 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 (Inveresk Research, 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.

 

Genetic Toxicity

Adequate information exists to characterise the mutagenicity of Rosin adduct esters. Results of bacterial mutation assays demonstrate that Resin acids and rosin acids, fumarated, esters with pentaerythritol and Rosin, fumarated, reaction products with glycerol and pentaerythritol were not mutagenic in four strains of Salmonella typhimurium and strain WP2 of Escherichia coli when tested in the absence or presence of exogenous metabolic activation. When tested using mammalian cells in vitro, in the absence and in the presence of S9 fraction, Resin acids and rosin acids, fumarated, esters with pentaerythritol was inactive in a gene mutation assay (L5178Y mouse lymphoma cells) and in a cytogenetics assay (human lymphocytes).

Reproductive / Developmental Toxicity

Reproductive Toxicity

 

Adequate information exists to characterise the reproductive toxicity of Rosin adduct esters. This includes results obtained from testing Resin acids and rosin acids, maleated, esters with pentaerythritol; Resin acids and rosin acids, fumarated, esters with glycerol; and Resin acids and rosin acids, fumarated, esters with pentaerythritol along with supporting data for Rosin, fumarated (the precursor of the fumarated ester), Rosin (the precursor of all substances included in this category) and Resin acids and rosin acids, esters with pentaerythritol (a Rosin reaction product). This information is summarised below.

 

In a key combined repeat dose toxicity study with reproduction/developmental toxicity screening test (Envigo Research Limited, 2017b), the test material (Resin acids and rosin acids, maleated, esters with pentaerythritol, CAS# 94581-17-6) was administered in the diet to three groups, each composed of twelve male and twelve female Wistar Han™:RccHan™:WIST strain rats, for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dietary concentrations of 3000, 7500 and 18000 ppm. The dietary concentration given to the high dietary concentration females during gestation and lactation was initially decreased to 15000 ppm to lessen the expected increase in achieved intake during these phases. However, due to adverse toxicity, the high dietary concentration was reduced to 12000 ppm for both sexes on study day 22. Estimated achieved dosages for males in Groups 2 to 4 during the study was 179.3, 441.9 and 783.4 mg/Kg bw/day respectively. For females, it was 221.5, 529.7 and 1508.1 mg/Kg bw/day during the pre-pairing phase, 237.0, 570.8 and 911.6 during gestation and 305.1, 610.9 and 794.0 mg/Kg bw/day during lactation respectively. A control group of twelve males and twelve females were treated with basal laboratory diet.

    

Based on the results this study the No Observed Adverse Effect Level (NOAEL) for systemic toxicity was considered to be 3000 ppm, principally due to effects on body weight gain, food consumption and adverse histopathological changes in the kidney and urinary bladder at 7500 ppm and 18000/12000 ppm. The NOAEL for reproduction was considered to be 7500 ppm due to lower corpora lutea count at 18000/12000 ppm. The NOAEL for survival, growth and development of the offspring was considered to be 3000 ppm due to effects on body weight, body weight gain and surface righting ability at 7500 ppm and 18000/12000 ppm.

 

In a second key combined repeated dose, reproductive/developmental toxicity study (Harlan Laboratories Ltd., 2014), the test material (Resin acids and rosin acids, fumarated, esters with glycerol, CAS# 97489-11-7) was administered by continuous dietary admixture to Wistar Han™:RccHan™:WIST strain rats (12/sex/concentration), for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dietary concentrations of 3000, 7500 or 18000 ppm (equivalent to a mean achieved dosage of 179.6, 432.2 and 995.8 mg/Kg bw/day respectively for males and 213.4, 544.4 and 1190.9 mg/Kg bw/day, respectively for females during the pre-pairing period). The dietary concentration given to the high dosage females during gestation and lactation was decreased to 15000 ppm to lessen the expected increase in achieved intake during these phases. A control group of twelve males and twelve females were treated with basal laboratory diet.

  

Although there were some statistically significant differences in treated animals from controls for the hematological parameters measured, these differences were considered not to be of toxicological significance. Blood chemical analysis however revealed reductions in alkaline phosphatase in animals of either sex treated with 18000/15000 or 7500 ppm, reductions in alanine aminotransferase in females treated with 7500 or 18000/15000 ppm, reductions in aspartate aminotransferase in females treated with 18000/15000 ppm. Females treated with 18000/15000 ppm also showed an increase in bilirubin whilst males treated with 18000 ppm also showed a reduction in albumin and albumin/globulin ratio. The majority of individual values (excluding albumin/globulin ratio) were outside of the expected normal range for rats of the strain and age used.

 

Microscopic examination revealed changes in the lungs, prostate and urinary bladder (females only). Histopathological examination of the lungs revealed an increase in incidence and severity of alveolar macrophages in animals of either sex treated with 18000/15000 ppm. Three of these females also showed white patches on the lungs at necropsy, which were confirmed as alveolar macrophages. At the low severity seen at this level, alveolar macrophages were not considered to be an adverse effect of treatment. Treatment-related urothelial hyperplasia and vacuolation was present in the urinary bladder of females treated with 18000/15000 ppm and an increase in incidence and severity of inflammation and lymphoid infiltrates was present in the prostate of males treated with 18000 or 7500 ppm. Whilst the effect on the prostate does not represent a degenerative change and there was no obvious effect on fertility (as assessed by mating success) an association with treatment cannot be excluded and thus does affect the reproductive NOEL.

 

Mating performance and fertility was unaffected by treatment. Offspring viability and litter size were also unaffected. Litter weights from females treated with 18000/15000 ppm were however reduced during lactation and mean offspring body weight and body weight gains were also reduced during lactation at this dosage. A relationship with reduced maternal body weight gain during gestation and lactation as a consequence of reduced food consumption cannot be excluded.

 

The ‘No Observed Adverse Effect Level’ (NOAEL) for systemic toxicity was therefore considered to be 7500 ppm for either sex. The NOAEL for reproductive toxicity was considered to be 3000 ppm.

 

In another Key combined repeated dose reproductive/developmental toxicity study (Harlan Laboratories Ltd., 2010), the test material (RARA, fumarated, esters with pentaerythritol, CAS# 94581-15-4) was administered by gavage to three groups, each of ten male and ten female Wistar Han™:HsdRccHan™:WIST strain rats, for up to eight weeks (including a two week maturation phase, pairing, gestation and early lactation for females), at dose levels of 30, 300 and 1000 mg/Kg/day. A control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP).

   

The oral administration of Resin acids and rosin acids, fumarated, esters with pentaerythritol to rats by gavage at a maximum dose level of 1000 mg/Kg/day resulted in treatment-related effects. The effects mainly consisted of microscopic changes in the urinary bladder. As such, a ‘No Observed Adverse Effect Level’ (NOAEL) was established at 300 mg/Kg/day for systemic toxicity. No treatment-related effects were detected for reproductive performance, hence a ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg/day.

 

Results for the supporting repeated dose/reproductive/developmental screening studies are presented below:

 

Rosin, fumarated was administered in the diet to rats at concentrations of 0, 1000 ppm (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 10,000 ppm (males 651-889 mg/kg bw/d; females 449-995 mg/kg bw/d) (Inveresk Research, 2004). 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. Food consumption and mean body weights were decreased in parental animals of both sexes at 10,000 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 parental (systemic) NOAEL. This is considered 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. With regard to reproductive parameters, there was a slight decrease in the mean number of implant sites per pregnancy and a consequent slight reduction in litter size at birth in the high dose group. A slight reduction in litter size between Day1-4 of lactation at 3000 ppm was due to the loss of most pups in one litter. As there were no effects of treatment on litter survival at 10,000 ppm the findings at 3000 ppm are considered to be incidental. Based on these results, the NOAEL for reproductive parameters was considered to be 10,000 ppm (males 651-889 mg/kg bw/d; females 449-995 mg/kg bw/d).

 

In a reproductive/developmental toxicity screening study, 10 rats/sex/group were exposed to Rosin at dietary concentrations of 0, 1000, 3000, or 10000 ppm for 41-45 days (females) or 30 days (males) in the diet (Inveresk Research, 2003a). Treatment with Rosin at 10000 ppm was associated with reduced weight gain/weight loss and reduced food consumption in the parental generation and a slight decrease in the mean number of implantation sites resulting in a subsequent slight reduction in litter size. Body weight gain reductions were also observed in males exposed to 3000 ppm Rosin. Adverse effects in the F1 pups were limited to slightly reduced litter and pup weights. Based on the results of the present study, the no-observed-effect-level (NOAEL) for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 3000 ppm for males and females.

 

In a reproductive/developmental toxicity screening study, 10 rats/sex/group were exposed ad libitum in the diet to Rosin pentaerythritol ester (Resin acids and rosin acids, esters with pentaerythritol) at dietary concentrations of 0, 1000, 5000, or 20000 ppm for 57-60 days (females) or 28 days (males) (Inveresk Research, 2003b). There were no test substance-related effects on reproductive performance of the parental females or on survival and development of the F1 pups. All findings occurred in a non dose-dependent manner, were spurious in nature, or were biologically irrelevant and were not considered related to rosin pentaerythritol ester consumption. The NOAEL for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 20000 ppm for males and females.

 

Developmental Toxicity

 

In a key developmental toxicity study (Envigo Research Laboratories, 2016b), the test material (Resin acids and Rosin acids, fumarated esters with Glycerol; CAS# 97489-11-7) was administered by continuous dietary admixture to three groups, each composed of twenty-four time mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between gestation days 3 and 19 (inclusive) at dietary concentrations of 3000, 7500, or 15000 ppm (equivalent to mean achieved dosages of 244.2, 622.2 or 1164.7 mg/kg bw/day, respectively). A further group of twenty-four time mated females was fed basal laboratory diet to serve as a control.

  

Dietary exposure to 15000 ppm of the test item was associated with lower maternal body weight gain during gestation and an initial effect on food consumption. While part of the lower overall weight gain observed was attributable to lower litter weight due to reduced foetal weight, an underlying effect on the pregnant dam was still present when body weight gain was adjusted for the contribution of the gravid uterus. For females at the 3000 and 7500 ppm dietary exposure levels, clinical signs, body weight performance, food consumption and macroscopic necropsy examinations did not indicate any obvious effect of treatment.

 

In-utero survival of the developing conceptus appeared unaffected by maternal dietary exposure to 15000 ppm of the test item with both pre-and post-implantation losses being comparable to control. This was despite a clear treatment-related reduction in foetal weight which resulted in lower litter weight at this dietary exposure level and which attained statistical significance and was significantly different than the other dietary exposure groups.

 

At the 7500 and 3000 ppm dietary exposure levels there was a reduction in foetal weight compared with controls. However, this finding was considered not to be adverse because the mean foetal weights were within the historical control range (2.42 - 4.97 g) and were within one standard deviation of the mean foetal weight of the control group. Furthermore, the difference in mean foetal weights between the low and intermediate dietary exposure groups was very marginal when compared with the difference in exposure levels, which is not suggestive of a clear indication of a relationship between maternal dietary exposure to 7500 and 3000 ppm of the test item and an effect on foetal weight.

 

Skeletal evaluation of foetuses from the 15000 ppm dietary exposure level showed significant differences compared to controls. The number of individual sites with reduced ossification and the difference in their incidence compared to controls was particularly higher, with a wide range of structures affected. Included within this were rib effects such as wavy rib. At the 7500 and 3000 ppm dietary exposure level, there was no increase in rib effects and the observed differences in ossification were limited to an increased incidence of incomplete ossification of one cranial bone (supra occipital) an no ossification of the metacarpals. The range of historical control incidence of foetuses with incomplete ossification of the supra occipital bone is 4.9%to 17.3% and the range of historical control incidence of foetuses with no ossification of the metacarpals is 5.3% to 22.2%, which is suggestive of high variability in the ossification of these structures at the end of gestation. Generalised delays have a common ‘finger print’, characterised by reduced ossification of bones that normally exhibit rapid ossification during the last few days of gestation (e.g. supra occipital bone and metacarpals) and denotes generalized growth delays with subsequent catch-up postnatally (Carney and Kimmel, 2007). Consequently, these isolated intergroup differences, in the absence of concomitant reductions in ossification of the other associated structures should be regarded as non-conclusive evidence of a foetal effect. Therefore, based on the findings in this study there was a clear difference in the effects observed between the 15000 ppm exposure level and the 7500 and 3000 ppm dietary exposure groups. The former clearly demonstrated a reduction in foetal weights and a significantly increased incidence of delays in skeletal development. At the 7500 and 3000 ppm dietary exposure levels the observations were within historical control incidence, of limited toxicological significance and were not considered to be indicative of adverse foetal effects.

 

The oral administration of the test material to pregnant rats by continuous dietary admixture from gestation Days 3 to 19, at a dietary concentration of 15000 ppm (equivalent to a mean achieved dosage of 1164.7 mg/kg bw/day) was associated with lower maternal body weight gain during gestation and an initial effect on food consumption. Therefore, the 7500 ppm dietary exposure level (equivalent to a mean achieved dosage of 622.2 mg/kg bw/day) was considered to represent the No Observed Adverse Effect Level (NOAEL) for the pregnant female.

 

In-utero survival of the developing conceptus was unaffected by maternal dietary exposure to 15000 ppm of the test item, although reduced foetal and placental weights and skeletal findings indicated an adverse effect on foetal growth. The NOAEL for developmental toxicity was therefore considered to be 7500 ppm (equivalent to a mean achieved dosage of 622.2 mg/kg bw/day).

 

In a supporting pre-natal developmental toxicity range-finding study (Envigo Research Laboratories, 2017c), the test material (RARA, fumarated, esters with glycerol (CAS# 97489-11-7)), was administered by continuous dietary admixture to three groups each of eight time-mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between Days 3 and 19 of gestation inclusive at dietary concentrations of 3000, 7500, and 15000 ppm (equivalent to a mean achieved dosage of 249.9, 617.7 and 1198.5 mg/kg bw/day respectively). A further group of eight time mated females was treated with basal laboratory diet to serve as a control. There was no effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio and pre-and post-implantation losses or on mean fetal, litter or placental weights at 3000, 7500 or 15000 ppm. There were no findings apparent for fetuses from treated females at external examination on Day 20 of gestation. The systemic and developmental toxicity NOAEL for RARA, fumarated esters with glycerol in rats is therefore determined to be 15000 ppm (equivalent to 1198.5 mg/Kg bw/day).

 

In a combined repeated dose, reproductive/developmental toxicity study (Harlan Laboratories Ltd, 2014), the test material (Resin acids and rosin acids, fumarated, esters with glycerol, CAS# 97489-11-7) was administered by continuous dietary admixture to Wistar Han™:RccHan™:WIST strain rats (12/sex/concentration), for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dietary concentrations of 3000, 7500 or 18000 ppm (equivalent to a mean achieved dosage of 179.6, 432.2 and 995.8 mg/kg bw/day respectively for males and 213.4, 544.4 and 1190.9 mg/kg bw/day, respectively for females during the pre-pairing period). The dietary concentration given to the high dosage females during gestation and lactation was decreased to 15000 ppm to lessen the expected increase in achieved intake during these phases. A control group of twelve males and twelve females were treated with basal laboratory diet.

 

Mating performance and fertility was unaffected by treatment. Offspring viability and litter size were also unaffected. Litter weights from females treated with 18000/15000 ppm were however reduced during lactation and mean offspring body weight and body weight gains were also reduced during lactation at this dosage. A relationship with reduced maternal body weight gain during gestation and lactation as a consequence of reduced food consumption cannot be excluded. The ‘No Observed Adverse Effect Level’ (NOAEL) for systemic toxicity was therefore considered to be 7500 ppm for either sex. The NOAEL for developmental toxicity was considered to be 7500 ppm, based on adverse treatment-related effects on offspring body weight observed at the highest concentration tested.

 

In a second combined repeated dose, reproductive/developmental toxicity study (Harlan Laboratories Ltd., 2010), the test material (Resin acids and rosin acids, fumarated, esters with pentaerythritol (CAS# 94581-15-4) was administered by gavage to three groups, each of ten male and ten female Wistar Han™:HsdRccHan™:WIST strain rats, for up to eight weeks (including a two week maturation phase, pairing, gestation and early lactation for females), at dose levels of 30, 300 and 1000 mg/kg/day. A control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP). No treatment-related effects were detected for reproductive performance, hence a ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg bw/day.

 

In a well-conducted reproductive and developmental toxicity screening study in rats (Inveresk Research, 2004), Rosin, fumarated was administered in the diet to rats at concentrations of 0, 1000 ppm (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 10,000 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. There was a slight decrease in the mean number of implant sites per pregnancy and a consequent slight reduction in litter size at birth in the high dose group. A slight reduction in litter size between Day1-4 of lactation at 3000 ppm was due to the loss of most pups in one litter. As there were no effects of treatment on litter survival at 10,000 ppm the findings at 3000 ppm are considered to be incidental. Based on these results, the NOAEL for reproductive parameters was considered to be 10,000 ppm (males 651-889 mg/Kg bw/d; females 449-995 mg/Kg bw/d).

 

In a reproductive/developmental toxicity screening study (Inveresk Research 2003a), 10 rats/sex/group were exposed ad libitum in the diet to the test material (Resin acids and rosin acids, esters with pentaerythritol; CAS# 8050-26-8) at dietary concentrations of 0, 1000, 5000, or 20000 ppm for 57-60 days (females) or 28 days (males). There were no test substance-related effects on reproductive performance of the parental females or on survival and development of the F1 pups. All findings occurred in a non dose-dependent manner, were spurious in nature, or were biologically irrelevant and were not considered related to rosin pentaerythritol ester consumption. The no observed adverse effect level (NOAEL) for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 20000 ppm for males and females, equivalent to received doses of 1864 mg/Kg bw/d and 1757 -2054 mg/Kg bw/d, respectively.  

 

In a reproductive/developmental toxicity screening study (Inveresk Research 2003b), 10 rats/sex/group were exposed to Gum Rosin (CAS# 8050-09-7) at dietary concentrations of 0, 1000, 3000, or 10000 ppm for 41-45 days (females) or 30 days (males) in the diet. Treatment with Gum Rosin at 10000 ppm was associated with reduced weight gain/weight loss and reduced food consumption in the parental generation and a slight decrease in the mean number of implantation sites resulting in a subsequent slight reduction in litter size. Body weight gain reductions were also observed in males exposed to 3000 ppm Gum Rosin. Adverse effects in the F1 pups were limited to slightly reduced litter and pup weights. Based on the results of the present study, the NOAEL for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 3000 ppm for males (equivalent to 248 mg/Kg bw/d) and females (equivalent to 309 mg/Kg bw/d).

DNEL Worker long-term dermal-systemic

Dose descriptor

A NOAEL of 209.1 mg/Kg bw/d will be used as the starting point.

Modification of dose descriptor

100% absorption after ingestion and 100% after skin contact are assumed.

Assessment factors (ECHA Guidance Chapter R8, Table R8-6, November 2012

 

Long-term DNEL Assessment Factors (Dermal)

Assessment Factor

Worker

Interspecies

2.5 (for systemic effects)

 

4 (Allometric scaling for rats)

Intraspecies

5 (for worker)

Exposure duration

2 (subchronic to chronic)

Issues related to reliability of the dose-response

1

Issues related to completeness and consistency of the available data

1

 

Overall AF

100

 

DNEL Worker long-term-systemic via dermal route = 209.1 / 100 = 2.09 mg/Kg bw/day

Endpoint for risk characterisation

Experimental  results  indicating  a  potential  to  cause  sensitisation  by  skin  contact,  with  associated classification  under  Regulation  (EC)  No.  1272/2008,  indicate  that  members  of  this  category  may present a risk to workers and the general population during normal handling and use. Qualitative risk characterisation (ECHA, 2010) will therefore be conducted on this endpoint.

General Population - Hazard via inhalation route

Systemic effects

Long term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
Acute/short term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
Acute/short term exposure
Hazard assessment conclusion:
hazard unknown but no further hazard information necessary as no exposure expected
DNEL related information

General Population - Hazard via dermal route

Systemic effects

Long term exposure
Hazard assessment conclusion:
DNEL (Derived No Effect Level)
Value:
1.046 mg/kg bw/day
Most sensitive endpoint:
repeated dose toxicity
Route of original study:
Oral
DNEL related information
DNEL derivation method:
ECHA REACH Guidance
Overall assessment factor (AF):
200
Dose descriptor starting point:
NOAEL
Value:
209.1 mg/kg bw/day
Modified dose descriptor starting point:
NOAEL
Value:
209.1 mg/kg bw/day
Explanation for the modification of the dose descriptor starting point:

Modification of dose descriptor:

 

Converted oral NOAEL rat (in mg/kg bw/day) into dermal NOAEL rat (in mg/kg bw/day) by correcting for differences in absorption between routes as well as for differences in dermal absorption between rats and humans:

 

corrected dermal NOAEL = oral NOAEL x (ABSoral-rat / ABSderm-rat) x (ABSderm-rat / ABSderm-human)

 

                                     = oral NOAEL x (ABSoral-rat / ABSderm-human)

 

                                     = 209.1 mg/Kg bw/day x (1 / 1) 

 

Note: Dermal absorption assumed not be higher than oral absorption, therefore no default factor (i.e. factor 1) introduced when performing oral-to-dermal extrapolation (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).

AF for dose response relationship:
1
Justification:
Default assessment factor when the starting point for the DNEL calculation is a NOAEL (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for differences in duration of exposure:
2
Justification:
Default assessment factor of 2 applied when extrapolating duration of exposure from sub-chronic to chronic (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for interspecies differences (allometric scaling):
4
Justification:
Allometric scaling factor for rats compared to humans (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for other interspecies differences:
2.5
Justification:
Additional factor of 2.5 for other interspecies differences; systemic effects (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for intraspecies differences:
10
Justification:
For general population, as standard procedure for threshold effects, a default assessment factor of 10 was applied (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for the quality of the whole database:
1
Justification:
Default assessment factor applied for good/standard quality of the database, taking into account completeness, consistency and the standard information requirements (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

Local effects

Long term exposure
Hazard assessment conclusion:
no hazard identified
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified

General Population - Hazard via oral route

Systemic effects

Long term exposure
Hazard assessment conclusion:
DNEL (Derived No Effect Level)
Value:
1.046 mg/kg bw/day
Most sensitive endpoint:
repeated dose toxicity
Route of original study:
Oral
DNEL related information
DNEL derivation method:
ECHA REACH Guidance
Overall assessment factor (AF):
200
Dose descriptor starting point:
NOAEL
Value:
209.1 mg/kg bw/day
Modified dose descriptor starting point:
NOAEL
Value:
209.1 mg/kg bw/day
Explanation for the modification of the dose descriptor starting point:

Modification of dose descriptor:

 

Converted oral NOAEL rat (in mg/Kg bw/day) into dermal NOAEL rat (in mg/Kg bw/day) by correcting for differences in absorption between routes as well as for differences in dermal absorption between rats and humans:

 

corrected dermal NOAEL = oral NOAEL x (ABSoral-rat / ABSderm-rat) x (ABSderm-rat / ABSderm-human)

 

                                     = oral NOAEL x (ABSoral-rat / ABSderm-human)

 

                                     = 209.1 mg/Kg bw/day x (1 / 1) 

 

Note: Dermal absorption assumed not be higher than oral absorption, therefore no default factor (i.e. factor 1) introduced when performing oral-to-dermal extrapolation (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).

AF for dose response relationship:
1
Justification:
Default assessment factor when the starting point for the DNEL calculation is a NOAEL (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for differences in duration of exposure:
2
Justification:
Default assessment factor of 2 applied when extrapolating duration of exposure from sub-chronic to chronic (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for interspecies differences (allometric scaling):
4
Justification:
Allometric scaling factor for rats compared to humans (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for other interspecies differences:
2.5
Justification:
Additional factor of 2.5 for other interspecies differences; systemic effects (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for intraspecies differences:
10
Justification:
For general population, as standard procedure for threshold effects, a default assessment factor of 10 was applied (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
AF for the quality of the whole database:
1
Justification:
Default assessment factor applied for good/standard quality of the database, taking into account completeness, consistency and the standard information requirements (ECHA Guidance on information requirements and chemical safety assessment Chapter R.8: Characterisation of dose [concentration]-response for human health, Version 2.1, November 2012).
Acute/short term exposure
Hazard assessment conclusion:
no hazard identified
DNEL related information

General Population - Hazard for the eyes

Local effects

Hazard assessment conclusion:
medium hazard (no threshold derived)

Additional information - General Population

Acute toxicity

ECHA Guidance R.8 (Chapter R.8.1.2.5) indicates that DNELs for acute toxicity are not required if no acute toxicity hazard leading to classification has been identified. The members of this category are not acutely toxic following oral or dermal exposure (LD50>2000 mg/Kg bw) while a low vapour pressure precludes inhalation exposure indicating a low of concern for this route of exposure. No DNELs for acute toxicity are therefore necessary.

 

Irritation

Corrosive and irritant effects on the skin and eye are local, concentration-dependent phenomena. However, while test results indicate that members of this category are irritating to eye (but not to skin), the nature of the data is such that no conclusion can be drawn with regard to any

dose-response relationship present. No DNEL for irritation can therefore be derived.

 

Sensitisation

The sensitisation potential of the substances included in this category is well understood and comprises results from six local lymph node assays and a guinea pig maximisation test. The LLNA results consistently showed evidence of a potential to induce skin sensitisation.

Intrinsic sensitising potency

ECHA Guidance R.8, Appendix R.8-10, (ECHA, 2010) states that while skin sensitisation is generally regarded as a threshold effect it may be very difficult to derive a threshold and to set a DNEL. Thus, the general approach for sensitisers involves a qualitative approach where a DNEL is used to judge any remaining/residual risks after the implementation of appropriate risk management measures (RMM) and occupational controls (OC).The extent of the RMM and OC required is dependent on the intrinsic sensitising potency of the substance.

 

For results obtained using the LLNA, intrinsic sensitising potency is based on the EC3 and defined (ECHA (2010), Appendix R.8-10) as follows:

 

Category

EC3 (%)

Extreme

<0.2%

Strong

>0.2 - <2

Moderate

>2

 

An EC3 of 14% was obtained with Resin acids and rosin acids, fumarated esters with pentaerythritol while three samples of Resin acids and rosin acids, maleated esters with pentaerythritol returned EC3 values of 16%, 17% and 22%. No EC3 could be derived from a LLNA test with a Resin acids and rosinacids, fumarated esters with glycerol and pentaerythritol since stimulation indices in a range 4.37-4.73 (obtained over a concentration range of 10-50% test substance) resulted in a negative intercept when the data were subject to linear regression analysis. Overall, however, these results indicate that membersof this category have a moderate potential to cause skin sensitisation.

 

Results obtained from a guinea pig maximisation test using an intradermal induction concentration of 7.5% indicate that Resin acids and rosin acids, fumarated esters with pentaerythritol was not a skin sensitiser. Although some anomalous results were obtained, the information summarised above indicates that the members of this category have a moderate potential to cause sensitisation following skin contact.

 

Derivation of a DNEL for sensitisation

ECHA Guidance R.8, Appendix R.8-10 (ECHA, 2010), indicates that the EC3 concentration from a LLNA test can be taken as a LOAEL for the induction of skin sensitisation (ECHA, 2010) after conversion into an equivalent dose per unit area of skin (µg/cm2). Assuming (i) a dose volume of 25 μL (according to the standard LLNA protocol); (ii) an estimated treatment area of 1 cm2 for the mouse ear; and (iii) an assumed density of is 1, the conversion is performed as follows:

 

EC3 [%] * 250 [µg/cm2/%] = EC3 [µg/cm2]

 

Based on an EC3 of 14% obtained for Resin acids and rosin acids, fumarated esters with pentaerythritol,the equivalent EC3 [µg/cm2] is therefore:

 

14% * 250 = 3500 µg/cm2

 

EC3 [µg/cm2] values of 4000, 4250 and 5500 µg/cm2were obtained from LLNA data for Resin acids and rosin acids, maleated esters with pentaerythritol.

 

The EC3 of 3500 µg/cm2 obtained for Resin acids and rosin acids, fumarated esters with pentaerythritol will be used to assess the magnitude of any remaining/residual risks after the use of RMMs and OCs recommended in the Qualitative Risk Assessment for this category. No Assessment Factors will be applied to this value since (relative to EC3 values available for other category members) it is already a conservative result, with skin penetration (and hence the capacity of the substance to induce skin sensitisation) enhanced in the LLNA test by the deliberate use of a solvent system that is not present in an occupational or consumer setting. The EC3 of 3500 µg/cm2 will therefore be used as a human NAEL (No Adverse Effect Level).

 

Repeated dose toxicity

 

Information exists to characterise the repeated dose toxicity of Rosin adduct esters. These are formed after modification of rosin with either fumaric acid or maleic anhydride followed by esterification with glycerol and/or pentaerythritol, and hence the adduct ester products therefore exhibit close structural similarities. The available data includes results obtained from testing Resin acids and rosin acids, fumarated, esters with glycerol; Resin acids and rosin acids, fumarated, esters with pentaerythritol; and Resin acids and rosin acids, maleated, esters with pentaerythritol with supporting information on Rosin, fumarated (precursor substance). This information is summarised below.

 

In a key oral repeated dose toxicity test conducted according to OECD Guideline 408 (Envigo Research Limited, 2017a), the test material (Resin acids and Rosin-acids, maleated, esters with pentaerythritol’ CAS# 94581-17-6) was administered in the diet to three groups, each composed of ten male and ten female Wistar Han™:RccHan™:WIST strain rats, for ninety consecutive days, at dietary concentrations of 3000, 6000 or 12000/15000 ppm (equivalent to mean achieved dosages of 209.1, 413.2 or 903.7 mg/Kg bw/day for males and 248.7, 436.7 or 1069.4 mg/Kg bw/day for females). The highest dietary level was increased from 12000 ppm to 15000 ppm after four weeks of treatment to maximise the achieved dosage for this dose group as the study progressed. A control group composed of ten males and ten females was fed basal laboratory diet.

  

No unscheduled mortality was observed through the study period and there were no obvious clinical effects of dietary exposure to test material at concentrations of 3000, 6000 or 12000/15000 ppm for either sex. Behavioural assessment, functional performance, and sensory reactivity assessments remained unaffected post treatment with the test material and there were no effects observed on food conversion efficiency, water consumption, or ophthalmoscopic parameters evaluated in animals of either sex. 

 

Dietary exposure to the test material at a concentration of 12000/15000 ppm was associated with adverse histopathological findings in the liver and urinary bladder. At a dietary exposure level of 6000 ppm, similar histopathological changes were apparent in both the liver and urinary bladder and, although the incidence was lower than observed at 12000/15000 ppm, these findings were considered to be adverse. At the 3000 ppm dietary exposure level, the adverse histopathological changes observed at higher exposure levels were not apparent and histopathological changes were limited to a low incidence of increased adipose tissue in the sternum bone marrow, which was considered to reflect individual variation and to be of no toxicological significance. Although occasional differences from control were observed for some blood chemistry parameters at the 3000 ppm level they were not regarded as adverse effects. Consequently, the No Observed Adverse Effect Level (NOAEL) of RARA, maleated, esters with pentaerythritol when administered via the diet for 90 consecutive days was considered to be 3000 ppm (equivalent to mean achieved dosages of 209.1 mg/kg bw/day for males and 248.7 mg/kg bw/day for females).

 

In a second key oral sub-chronic toxicity study (Envigo Research Limited, 2016a), the test material (Resin acids and Rosin acids, fumarated, esters with pentaerythitol; CAS# 94581-15-4) was administered by continuous dietary exposure to Wistar Han™:RccHan™:WIST strain rats (10/sex/concentration), for ninety consecutive days, at dietary concentrations of 3000 and 6000 ppm for the low and intermediate concentration groups (equivalent to a mean achieved dosage of 210.0 and 414.1 mg/Kg bw/day for males and 262.4 and 511.2 mg/Kg bw/day for females). For the high concentration group, rats were initially fed diet containing 12000 ppm for two weeks followed by 15000 ppm for four weeks and subsequently 18000 ppm for the remainder of the study (equivalent to a mean achieved dosage of 1090.0 mg/Kg bw/day for males and 1298.9 mg/Kg bw/day for females). A control group of ten males and ten females were treated with basal laboratory diet.

  

The continuous oral (dietary) administration of the test material for ninety consecutive days, did not result in any toxicologically significant effects. Therefore, based on the lack of adverse treatment-related effects observed through the study period, the No Observed Adverse Effect Level (NOAEL) for systemic toxicity for both sexes was determined to be 18000 ppm (equivalent to a mean achieved dosage of 1090.0 mg/Kg bw/day for males and 1298.9 mg/Kg bw/day for females).

 

In another key oral sub-chronic toxicity study (Envigo Research Limited, 2015a), the test material (RARA, fumarated, esters with glycerol; CAS# 97489-11-7) was administered by continuous dietary admixture Wistar Han™:RccHan™:WIST strain rats (10/sex/concentration), for ninety consecutive days, at nominal dietary concentrations of 3000, 7500 and 18000 ppm (equivalent to mean achieved dosages of 211.9, 574.0 and 1296.1 mg/Kg bw/day for males and 248.1, 598.9 and 1482.2 mg/Kg bw/day for females). A control group of ten males and ten females were treated with basal laboratory diet.

 

Continuous oral dietary administration of the test material resulted in reduced body weight gain in males treated with 18000 ppm. There were no clinical signs evident for animals of either sex treated with 3000, 7500 or 18000 ppm. The body weight development of high dose males was reduced relative to controls during Weeks 1, 3, 4 and 7 of treatment. Subsequently a 10% reduction in overall body weight gain was evident in these males.No consistent effect on body weight gain was evident in females and no adverse effect was evident in food consumption for either sex. Therefore, the effect on body weight gain in males was most likely associated with test item toxicity rather than a palatability effect.

 

Some statistically significant differences in hematological and blood chemistry parameters were observed in treated animals relative to controls. However, majority of individual values were within the expected normal ranges for rats of the strain and age used. Therefore, these differences were not considered to be toxicologically significant. Macroscopic and microscopic examinations did not reveal any treatment related effects in treated animals.

 

Based on the results of this study, the No Observed Adverse Effect Level (NOAEL) for RARA, fumarated, esters with glycerol was determined to be 18000 ppm for female rats and 7500 ppm for male rats.

 

Resin acids and rosin acids, fumarated, esters with glycerol (CAS# 97489-11-7) was administered to rat by gavage at concentrations of 3000, 7500, or 18000/15000 ppm (Harlan Laboratories Ltd., 2014). The result of the study showed microscopic lung changes in animals of either sex treated with 18000/15000 ppm, microscopic urinary bladder changes in females treated with 18000/15000 ppm and microscopic prostate changes in males 18000 or 7500 ppm. The ‘No Observed Effect Level (NOEL) for systemic toxicity was considered to be 7500 ppm for either sex. 

 

Resin acids and rosin acids, fumarated, esters with pentaerythritol was administered by oral gavage to male and female rats at treatments levels of 0, 30, 300 or 1000 mg/Kg bw/g for approximately 6-7 weeks (Harlan Laboratories Ltd, 2010b). Treatment-related changes in bladder (comprising epithelial hyperplasia, ulceration and erosion with associated subepithelial oedema and inflammation) seen at 1000 mg/Kg/day but not at any other treatment level. The condition was more prevalent among females. A NOAEL of 300 mg/Kg/day for systemic toxicity was established by this study.

 

In a supporting study, Rosin, fumarated (the precursor of Rosin, fumarated, esters with pentaerythritol) 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 (Inveresk Research, 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.

 

Genetic Toxicity

Adequate information exists to characterise the mutagenicity of Rosin adduct esters. Results of bacterial mutation assays demonstrate that Resin acids and rosin acids, fumarated, esters with pentaerythritol and Rosin, fumarated, reaction products with glycerol and pentaerythritol were not mutagenic in four strains of Salmonella typhimurium and strain WP2 of Escherichia coli when tested in the absence or presence of exogenous metabolic activation. When tested using mammalian cells in vitro, in the absence and in the presence of S9 fraction, Resin acids and rosin acids, fumarated, esters with pentaerythritol was inactive in a gene mutation assay (L5178Y mouse lymphoma cells) and in a cytogenetics assay (human lymphocytes).

Reproductive / Developmental Toxicity

Reproductive Toxicity

 

Adequate information exists to characterise the reproductive toxicity of Rosin adduct esters. This includes results obtained from testing Resin acids and rosin acids, maleated, esters with pentaerythritol; Resin acids and rosin acids, fumarated, esters with glycerol; and Resin acids and rosin acids, fumarated, esters with pentaerythritol along with supporting data for Rosin, fumarated (the precursor of the fumarated ester), Rosin (the precursor of all substances included in this category) and Resin acids and rosin acids, esters with pentaerythritol (a Rosin reaction product). This information is summarised below.

 

In a key combined repeat dose toxicity study with reproduction/developmental toxicity screening test (Envigo Research Limited, 2017b), the test material (Resin acids and rosin acids, maleated, esters with pentaerythritol, CAS# 94581-17-6) was administered in the diet to three groups, each composed of twelve male and twelve female Wistar Han™:RccHan™:WIST strain rats, for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dietary concentrations of 3000, 7500 and 18000 ppm. The dietary concentration given to the high dietary concentration females during gestation and lactation was initially decreased to 15000 ppm to lessen the expected increase in achieved intake during these phases. However, due to adverse toxicity, the high dietary concentration was reduced to 12000 ppm for both sexes on study day 22. Estimated achieved dosages for males in Groups 2 to 4 during the study was 179.3, 441.9 and 783.4 mg/Kg bw/day respectively. For females, it was 221.5, 529.7 and 1508.1 mg/Kg bw/day during the pre-pairing phase, 237.0, 570.8 and 911.6 during gestation and 305.1, 610.9 and 794.0 mg/Kg bw/day during lactation respectively. A control group of twelve males and twelve females were treated with basal laboratory diet.

    

Based on the results this study the No Observed Adverse Effect Level (NOAEL) for systemic toxicity was considered to be 3000 ppm, principally due to effects on body weight gain, food consumption and adverse histopathological changes in the kidney and urinary bladder at 7500 ppm and 18000/12000 ppm. The NOAEL for reproduction was considered to be 7500 ppm due to lower corpora lutea count at 18000/12000 ppm. The NOAEL for survival, growth and development of the offspring was considered to be 3000 ppm due to effects on body weight, body weight gain and surface righting ability at 7500 ppm and 18000/12000 ppm.

 

In a second key combined repeated dose, reproductive/developmental toxicity study (Harlan Laboratories Ltd., 2014), the test material (Resin acids and rosin acids, fumarated, esters with glycerol, CAS# 97489-11-7) was administered by continuous dietary admixture to Wistar Han™:RccHan™:WIST strain rats (12/sex/concentration), for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dietary concentrations of 3000, 7500 or 18000 ppm (equivalent to a mean achieved dosage of 179.6, 432.2 and 995.8 mg/Kg bw/day respectively for males and 213.4, 544.4 and 1190.9 mg/Kg bw/day, respectively for females during the pre-pairing period). The dietary concentration given to the high dosage females during gestation and lactation was decreased to 15000 ppm to lessen the expected increase in achieved intake during these phases. A control group of twelve males and twelve females were treated with basal laboratory diet.

  

Although there were some statistically significant differences in treated animals from controls for the hematological parameters measured, these differences were considered not to be of toxicological significance. Blood chemical analysis however revealed reductions in alkaline phosphatase in animals of either sex treated with 18000/15000 or 7500 ppm, reductions in alanine aminotransferase in females treated with 7500 or 18000/15000 ppm, reductions in aspartate aminotransferase in females treated with 18000/15000 ppm. Females treated with 18000/15000 ppm also showed an increase in bilirubin whilst males treated with 18000 ppm also showed a reduction in albumin and albumin/globulin ratio. The majority of individual values (excluding albumin/globulin ratio) were outside of the expected normal range for rats of the strain and age used.

 

Microscopic examination revealed changes in the lungs, prostate and urinary bladder (females only). Histopathological examination of the lungs revealed an increase in incidence and severity of alveolar macrophages in animals of either sex treated with 18000/15000 ppm. Three of these females also showed white patches on the lungs at necropsy, which were confirmed as alveolar macrophages. At the low severity seen at this level, alveolar macrophages were not considered to be an adverse effect of treatment. Treatment-related urothelial hyperplasia and vacuolation was present in the urinary bladder of females treated with 18000/15000 ppm and an increase in incidence and severity of inflammation and lymphoid infiltrates was present in the prostate of males treated with 18000 or 7500 ppm. Whilst the effect on the prostate does not represent a degenerative change and there was no obvious effect on fertility (as assessed by mating success) an association with treatment cannot be excluded and thus does affect the reproductive NOEL.

 

Mating performance and fertility was unaffected by treatment. Offspring viability and litter size were also unaffected. Litter weights from females treated with 18000/15000 ppm were however reduced during lactation and mean offspring body weight and body weight gains were also reduced during lactation at this dosage. A relationship with reduced maternal body weight gain during gestation and lactation as a consequence of reduced food consumption cannot be excluded.

 

The ‘No Observed Adverse Effect Level’ (NOAEL) for systemic toxicity was therefore considered to be 7500 ppm for either sex. The NOAEL for reproductive toxicity was considered to be 3000 ppm.

 

In another Key combined repeated dose reproductive/developmental toxicity study (Harlan Laboratories Ltd., 2010), the test material (RARA, fumarated, esters with pentaerythritol, CAS# 94581-15-4) was administered by gavage to three groups, each of ten male and ten female Wistar Han™:HsdRccHan™:WIST strain rats, for up to eight weeks (including a two week maturation phase, pairing, gestation and early lactation for females), at dose levels of 30, 300 and 1000 mg/Kg/day. A control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP).

   

The oral administration of Resin acids and rosin acids, fumarated, esters with pentaerythritol to rats by gavage at a maximum dose level of 1000 mg/Kg/day resulted in treatment-related effects. The effects mainly consisted of microscopic changes in the urinary bladder. As such, a ‘No Observed Adverse Effect Level’ (NOAEL) was established at 300 mg/Kg/day for systemic toxicity. No treatment-related effects were detected for reproductive performance, hence a ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg/day.

 

Results for the supporting repeated dose/reproductive/developmental screening studies are presented below:

 

Rosin, fumarated was administered in the diet to rats at concentrations of 0, 1000 ppm (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 10,000 ppm (males 651-889 mg/kg bw/d; females 449-995 mg/kg bw/d) (Inveresk Research, 2004). 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. Food consumption and mean body weights were decreased in parental animals of both sexes at 10,000 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 parental (systemic) NOAEL. This is considered 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. With regard to reproductive parameters, there was a slight decrease in the mean number of implant sites per pregnancy and a consequent slight reduction in litter size at birth in the high dose group. A slight reduction in litter size between Day1-4 of lactation at 3000 ppm was due to the loss of most pups in one litter. As there were no effects of treatment on litter survival at 10,000 ppm the findings at 3000 ppm are considered to be incidental. Based on these results, the NOAEL for reproductive parameters was considered to be 10,000 ppm (males 651-889 mg/kg bw/d; females 449-995 mg/kg bw/d).

 

In a reproductive/developmental toxicity screening study, 10 rats/sex/group were exposed to Rosin at dietary concentrations of 0, 1000, 3000, or 10000 ppm for 41-45 days (females) or 30 days (males) in the diet (Inveresk Research, 2003a). Treatment with Rosin at 10000 ppm was associated with reduced weight gain/weight loss and reduced food consumption in the parental generation and a slight decrease in the mean number of implantation sites resulting in a subsequent slight reduction in litter size. Body weight gain reductions were also observed in males exposed to 3000 ppm Rosin. Adverse effects in the F1 pups were limited to slightly reduced litter and pup weights. Based on the results of the present study, the no-observed-effect-level (NOAEL) for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 3000 ppm for males and females.

 

In a reproductive/developmental toxicity screening study, 10 rats/sex/group were exposed ad libitum in the diet to Rosin pentaerythritol ester (Resin acids and rosin acids, esters with pentaerythritol) at dietary concentrations of 0, 1000, 5000, or 20000 ppm for 57-60 days (females) or 28 days (males) (Inveresk Research, 2003b). There were no test substance-related effects on reproductive performance of the parental females or on survival and development of the F1 pups. All findings occurred in a non dose-dependent manner, were spurious in nature, or were biologically irrelevant and were not considered related to rosin pentaerythritol ester consumption. The NOAEL for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 20000 ppm for males and females.

 

Developmental Toxicity

 

In a key developmental toxicity study (Envigo Research Laboratories, 2016b), the test material (Resin acids and Rosin acids, fumarated esters with Glycerol; CAS# 97489-11-7) was administered by continuous dietary admixture to three groups, each composed of twenty-four time mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between gestation days 3 and 19 (inclusive) at dietary concentrations of 3000, 7500, or 15000 ppm (equivalent to mean achieved dosages of 244.2, 622.2 or 1164.7 mg/kg bw/day, respectively). A further group of twenty-four time mated females was fed basal laboratory diet to serve as a control.

  

Dietary exposure to 15000 ppm of the test item was associated with lower maternal body weight gain during gestation and an initial effect on food consumption. While part of the lower overall weight gain observed was attributable to lower litter weight due to reduced foetal weight, an underlying effect on the pregnant dam was still present when body weight gain was adjusted for the contribution of the gravid uterus. For females at the 3000 and 7500 ppm dietary exposure levels, clinical signs, body weight performance, food consumption and macroscopic necropsy examinations did not indicate any obvious effect of treatment.

 

In-utero survival of the developing conceptus appeared unaffected by maternal dietary exposure to 15000 ppm of the test item with both pre-and post-implantation losses being comparable to control. This was despite a clear treatment-related reduction in foetal weight which resulted in lower litter weight at this dietary exposure level and which attained statistical significance and was significantly different than the other dietary exposure groups.

 

At the 7500 and 3000 ppm dietary exposure levels there was a reduction in foetal weight compared with controls. However, this finding was considered not to be adverse because the mean foetal weights were within the historical control range (2.42 - 4.97 g) and were within one standard deviation of the mean foetal weight of the control group. Furthermore, the difference in mean foetal weights between the low and intermediate dietary exposure groups was very marginal when compared with the difference in exposure levels, which is not suggestive of a clear indication of a relationship between maternal dietary exposure to 7500 and 3000 ppm of the test item and an effect on foetal weight.

 

Skeletal evaluation of foetuses from the 15000 ppm dietary exposure level showed significant differences compared to controls. The number of individual sites with reduced ossification and the difference in their incidence compared to controls was particularly higher, with a wide range of structures affected. Included within this were rib effects such as wavy rib. At the 7500 and 3000 ppm dietary exposure level, there was no increase in rib effects and the observed differences in ossification were limited to an increased incidence of incomplete ossification of one cranial bone (supra occipital) an no ossification of the metacarpals. The range of historical control incidence of foetuses with incomplete ossification of the supra occipital bone is 4.9%to 17.3% and the range of historical control incidence of foetuses with no ossification of the metacarpals is 5.3% to 22.2%, which is suggestive of high variability in the ossification of these structures at the end of gestation. Generalised delays have a common ‘finger print’, characterised by reduced ossification of bones that normally exhibit rapid ossification during the last few days of gestation (e.g. supra occipital bone and metacarpals) and denotes generalized growth delays with subsequent catch-up postnatally (Carney and Kimmel, 2007). Consequently, these isolated intergroup differences, in the absence of concomitant reductions in ossification of the other associated structures should be regarded as non-conclusive evidence of a foetal effect. Therefore, based on the findings in this study there was a clear difference in the effects observed between the 15000 ppm exposure level and the 7500 and 3000 ppm dietary exposure groups. The former clearly demonstrated a reduction in foetal weights and a significantly increased incidence of delays in skeletal development. At the 7500 and 3000 ppm dietary exposure levels the observations were within historical control incidence, of limited toxicological significance and were not considered to be indicative of adverse foetal effects.

 

The oral administration of the test material to pregnant rats by continuous dietary admixture from gestation Days 3 to 19, at a dietary concentration of 15000 ppm (equivalent to a mean achieved dosage of 1164.7 mg/kg bw/day) was associated with lower maternal body weight gain during gestation and an initial effect on food consumption. Therefore, the 7500 ppm dietary exposure level (equivalent to a mean achieved dosage of 622.2 mg/kg bw/day) was considered to represent the No Observed Adverse Effect Level (NOAEL) for the pregnant female.

 

In-utero survival of the developing conceptus was unaffected by maternal dietary exposure to 15000 ppm of the test item, although reduced foetal and placental weights and skeletal findings indicated an adverse effect on foetal growth. The NOAEL for developmental toxicity was therefore considered to be 7500 ppm (equivalent to a mean achieved dosage of 622.2 mg/kg bw/day).

 

In a supporting pre-natal developmental toxicity range-finding study (Envigo Research Laboratories, 2017c), the test material (RARA, fumarated, esters with glycerol (CAS# 97489-11-7)), was administered by continuous dietary admixture to three groups each of eight time-mated Sprague-Dawley Crl:CD® (SD) IGS BR strain rats, between Days 3 and 19 of gestation inclusive at dietary concentrations of 3000, 7500, and 15000 ppm (equivalent to a mean achieved dosage of 249.9, 617.7 and 1198.5 mg/kg bw/day respectively). A further group of eight time mated females was treated with basal laboratory diet to serve as a control. There was no effect of maternal treatment on litter data as assessed by numbers of implantations, in-utero offspring survival (as assessed by the mean numbers of early or late resorptions), live litter size, sex ratio and pre-and post-implantation losses or on mean fetal, litter or placental weights at 3000, 7500 or 15000 ppm. There were no findings apparent for fetuses from treated females at external examination on Day 20 of gestation. The systemic and developmental toxicity NOAEL for RARA, fumarated esters with glycerol in rats is therefore determined to be 15000 ppm (equivalent to 1198.5 mg/Kg bw/day).

 

In a combined repeated dose, reproductive/developmental toxicity study (Harlan Laboratories Ltd, 2014), the test material (Resin acids and rosin acids, fumarated, esters with glycerol, CAS# 97489-11-7) was administered by continuous dietary admixture to Wistar Han™:RccHan™:WIST strain rats (12/sex/concentration), for up to eight weeks (including a two week pre-pairing phase, pairing, gestation and early lactation for females), at dietary concentrations of 3000, 7500 or 18000 ppm (equivalent to a mean achieved dosage of 179.6, 432.2 and 995.8 mg/kg bw/day respectively for males and 213.4, 544.4 and 1190.9 mg/kg bw/day, respectively for females during the pre-pairing period). The dietary concentration given to the high dosage females during gestation and lactation was decreased to 15000 ppm to lessen the expected increase in achieved intake during these phases. A control group of twelve males and twelve females were treated with basal laboratory diet.

 

Mating performance and fertility was unaffected by treatment. Offspring viability and litter size were also unaffected. Litter weights from females treated with 18000/15000 ppm were however reduced during lactation and mean offspring body weight and body weight gains were also reduced during lactation at this dosage. A relationship with reduced maternal body weight gain during gestation and lactation as a consequence of reduced food consumption cannot be excluded. The ‘No Observed Adverse Effect Level’ (NOAEL) for systemic toxicity was therefore considered to be 7500 ppm for either sex. The NOAEL for developmental toxicity was considered to be 7500 ppm, based on adverse treatment-related effects on offspring body weight observed at the highest concentration tested.

 

In a second combined repeated dose, reproductive/developmental toxicity study (Harlan Laboratories Ltd., 2010), the test material (Resin acids and rosin acids, fumarated, esters with pentaerythritol (CAS# 94581-15-4) was administered by gavage to three groups, each of ten male and ten female Wistar Han™:HsdRccHan™:WIST strain rats, for up to eight weeks (including a two week maturation phase, pairing, gestation and early lactation for females), at dose levels of 30, 300 and 1000 mg/kg/day. A control group of ten males and ten females was dosed with vehicle alone (Arachis oil BP). No treatment-related effects were detected for reproductive performance, hence a ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg bw/day.

 

In a well-conducted reproductive and developmental toxicity screening study in rats (Inveresk Research, 2004), Rosin, fumarated was administered in the diet to rats at concentrations of 0, 1000 ppm (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 10,000 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. There was a slight decrease in the mean number of implant sites per pregnancy and a consequent slight reduction in litter size at birth in the high dose group. A slight reduction in litter size between Day1-4 of lactation at 3000 ppm was due to the loss of most pups in one litter. As there were no effects of treatment on litter survival at 10,000 ppm the findings at 3000 ppm are considered to be incidental. Based on these results, the NOAEL for reproductive parameters was considered to be 10,000 ppm (males 651-889 mg/Kg bw/d; females 449-995 mg/Kg bw/d).

 

In a reproductive/developmental toxicity screening study (Inveresk Research 2003a), 10 rats/sex/group were exposed ad libitum in the diet to the test material (Resin acids and rosin acids, esters with pentaerythritol; CAS# 8050-26-8) at dietary concentrations of 0, 1000, 5000, or 20000 ppm for 57-60 days (females) or 28 days (males). There were no test substance-related effects on reproductive performance of the parental females or on survival and development of the F1 pups. All findings occurred in a non dose-dependent manner, were spurious in nature, or were biologically irrelevant and were not considered related to rosin pentaerythritol ester consumption. The no observed adverse effect level (NOAEL) for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 20000 ppm for males and females, equivalent to received doses of 1864 mg/Kg bw/d and 1757 -2054 mg/Kg bw/d, respectively.  

 

In a reproductive/developmental toxicity screening study (Inveresk Research 2003b), 10 rats/sex/group were exposed to Gum Rosin (CAS# 8050-09-7) at dietary concentrations of 0, 1000, 3000, or 10000 ppm for 41-45 days (females) or 30 days (males) in the diet. Treatment with Gum Rosin at 10000 ppm was associated with reduced weight gain/weight loss and reduced food consumption in the parental generation and a slight decrease in the mean number of implantation sites resulting in a subsequent slight reduction in litter size. Body weight gain reductions were also observed in males exposed to 3000 ppm Gum Rosin. Adverse effects in the F1 pups were limited to slightly reduced litter and pup weights. Based on the results of the present study, the NOAEL for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 3000 ppm for males (equivalent to 248 mg/Kg bw/d) and females (equivalent to 309 mg/Kg bw/d).

DNEL General Population long-term dermal-systemic

Dose descriptor

A NOAEL of 209.1 mg/Kg bw/d will be used as the starting point.

Modification of dose descriptor

100% absorption after ingestion and 100% after skin contact are assumed.

Assessment factors (ECHA Guidance Chapter R8, Table R8-6, November 2012

 

Long-term DNEL Assessment Factors (Dermal)

Assessment Factor

General Population

Interspecies

2.5 (for systemic effects)

 

4 (Allometric scaling for rats)

Intraspecies

10 (for general population)

Exposure duration

2 (subchronic to chronic)

Issues related to reliability of the dose-response

1

Issues related to completeness and consistency of the available data

1

 

Overall AF

200

DNEL General Population long-term systemic-via dermal route = 209.1 / 200 = 1.0455 mg/Kg bw/day

DNEL General Population long-term oral-systemic

Dose descriptor

A NOAEL of 213.1 mg/Kg bw/d will be used as the starting point.

Modification of dose descriptor

100% absorption after ingestion and 100% after skin contact are assumed.

Assessment factors (ECHA Guidance Chapter R8, Table R8-6, November 2012

 

Long-term DNEL Assessment Factors (Oral)

Assessment Factor

General Population

Interspecies

2.5 (for systemic effects)

 

4 (Allometric scaling for rats)

Intraspecies

10 (for general population)

Exposure duration

2 (subchronic to chronic)

Issues related to reliability of the dose-response

1

Issues related to completeness and consistency of the available data

1

 

Overall AF

200

 

DNEL General Population long-term-systemic via oral route = 209.1 / 200 = 1.0455 mg/Kg bw/day

Endpoint for risk characterisation

Experimental  results  indicating  a  potential  to  cause  sensitisation  by  skin  contact,  with  associated classification  under  Regulation  (EC)  No.  1272/2008,  indicate  that  members  of  this  category  may present a risk to workers and the general population during normal handling and use. Qualitative risk characterisation (ECHA, 2010) will therefore be conducted on this endpoint.