Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Three key Guideline (OECD 422) reproductive/developmental toxicity studies were identified for 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.

1) Resin acids and rosin acids, maleated, esters with pentaerythritol, CAS# 94581-17-6): The No Observed Adverse Effect Level (NOAEL) for reproduction was considered to be 7500 ppm (Mean Achieved Dosage Level equivalent to 441.9 mg/Kg bw/day in males and 529.7 mg/Kg bw/day in females) due to lower corpora lutea count at 18000/12000 ppm.

 

2) Resin acids and rosin acids, fumarated, esters with glycerol (CAS# 97489-11-7): The NOAEL for reproductive toxicity was considered to be 3000 ppm, (equivalent to a mean achieved dosage of 179.6 mg/Kg bw/day in males and 213.4 mg/Kg bw/day), based on an increase in incidence and severity of inflammation and lymphoid infiltrates was present in the prostate of males treated with 18000 or 7500 ppm.

  

3) Resin acids and rosin acids, fumarated, esters with pentaerythritol (CAS# 94581-15-4): No treatment-related effects were detected for reproductive performance, hence the ‘No Observed Effect Level’ (NOEL) for reproductive toxicity was considered to be 1000 mg/Kg/day.

Supporting data is available 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).

For Rosin, fumarated (CAS# 65997-04-8), there was no clear evidence of test substance-related effects on reproduction of F0 males and females or on survival and development of F1 pups. Therefore, the NOAEL for reproductive parameters was considered to be 10,000 ppm (males 651-889 mg/Kg bw/day; females 449-995 mg/Kg bw/day), the highest concentration tested.

 

For Resin acids and rosin acids, esters with pentaerythritol (CAS# 8050-26-8), the no observed adverse effect level (NOAEL) for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 20000 ppm (the highest concentration tested) for males and females (equivalent to received doses of 1864 mg/Kg bw/day and 1757-2054 mg/Kg bw/day, respectively).  

 

For Rosin (CAS# 8050-09-7), the NOAEL for reproductive toxicity in Sprague-Dawley rats was considered to be 3000 ppm (equivalent to 248 mg/Kg bw/day in males and 309 mg/Kg bw/day in females), based on slightly decreased mean number of implantation sites per pregnancy resulting in a subsequent slight reduction in litter size in dams exposed to 10000 ppm.

Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

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.

 

Clinical signs, behavioural assessments, body weight change and food and water consumption were monitored during the study. Pairing of animals within each dose group was undertaken on a one male: one female basis within each treatment group on study day 15, with females subsequently being allowed to litter and rear their offspring to lactation day 5. During the lactation phase, daily clinical observations were performed on all surviving offspring, together with litter size and offspring weights and assessment of surface righting reflex. Extensive functional observations were performed on five males from each dose group after the completion of the pairing phase, and for five parental females from each dose group on day 4 post-partum. Haematology and blood chemistry were evaluated prior to termination on five males and females from each dose group. Surviving adult males were terminated on study day 43/44, followed by the termination of all surviving females and offspring on day 5 post-partum. All adult animals, including decedents were subjected to a gross necropsy examination and histopathological evaluation of selected tissues was performed.

 

There were three unscheduled and treatment-related deaths at the highest dietary level (18000 ppm). No signs of treatment-related clinical toxicity were apparent in any of the surviving animals at 3000, 7500 and 18000/ 12000 ppm. Behavioural assessments, functional performance, sensory reactivity assessments, and haematology parameters were unaffected by dietary exposure at 3000, 7500 and 18000/ 12000 ppm. Gross necropsy did not reveal any remarkable findings in rats of either sex.

 

At the 18000 ppm dietary concentration, notable mean body weight loss during the first week of dietary exposure and statistically significant lower body weight gain during Week 2, compared with control, was observed in males. During Week 3, a number of individual animals showed marked body weight loss, resulting in a mean bodyweight loss and the dietary concentration was lowered to 12000 ppm for the high dietary level group. No further mean body weight losses or statistically significant differences in body weight gain were apparent for the remainder of the study although the overall body weight gain at termination of the study remained statistically significantly lower than control. Females in the 18000 ppm group, showed notable mean body weight loss during the first week of dietary exposure but recovery of body weight gain was apparent during the following week. Body weight gain of females at 18000/12000 ppm was lower than control throughout gestation, particularly during the last week of gestation. During lactation, marginal mean body weight loss was apparent for females in the 18000/12000 ppm group compared with control. Body weight gains for males receiving diets containing 7500 ppm were generally lower than control throughout the study and statistically significant lower overall body weight gain was apparent for these animals at the end of the study. Body weight gains for females that received diets containing 7500 ppm were lower than control during the two week pre-pairing period and throughout gestation and lactation, although differences only attained statistical significance during the second week of gestation and during lactation. There was no obvious effect of dietary exposure to 3000 ppm on body weight gains for both sexes throughout the study.

 

At the 18000 ppm dietary concentration, males showed notably lower mean food consumption during the first and, to a lesser extent, second week of dietary exposure compared with control. Following the pairing period, food consumption remained lower than control, despite the animals only receiving 12000 ppm at this stage of the study. For females at the 18000/12000 ppm dietary concentration, food consumption was lower than control throughout gestation and lactation phases; these differences were particularly noticeable during late gestation and during lactation. At the 7500 ppm dietary concentration, food consumption for males was slightly lower than control during the post-pairing period. Food consumption for females was lower than control throughout gestation, and, to a greater extent, lactation. At the 3000 ppm dietary concentration, food consumption for each sex was considered to have been unaffected by dietary exposure to the test material.

 

At the 18000 ppm dietary concentration, food conversion efficiency was lower for both sexes during the first week of dietary exposure to the test item. At the 3000 and 7500 ppm dietary concentrations, food conversion efficiency for both sexes appeared to be unaffected by dietary exposure to the test material.

 

At the 18000 ppm dietary concentration, males showed lower mean water consumption compared with control, throughout the two week pre-pairing period. For males at the 7500 ppm dietary concentration, water intake during the pre-pairing period appeared lower than control from day 11 of the study; prior to this, values did not indicate any consistent effect of dietary exposure. For males at the 3000 ppm dietary concentration and females at all dietary levels, water consumption during the pre-pairing period was unaffected by dietary exposure.

 

For both sexes at all dietary concentrations, statistically significant higher levels of total bilirubin were observed when compared with control, with mean values showing a consistent relationship to test material exposure. The majority of individual valuesat the 7500 and 18000/12000 ppm dietary concentrations exceeded the historical control range. All individual values for the 3000 ppm dietary group were within the historical control range. For females at the 7500 and 18000/12000 ppm dietary concentrations, statistically significant lower total protein, albumin and albumin/globulin ratio were observed compared with control; however the majority of individual values were within the historical control range. Statistically significant lower total protein and albumin levels were also observed for males at the 18000/12000 ppm dietary concentrations, however, the majority of individual values were within the historical control range. For females at the 7500 ppm dietary level and both sexes at the 18000/12000 ppm dietary level, alkaline phosphatase was lower than control with differences attaining statistical significance; the majority of individual values for these exposed females were below the historical control range but all values for these exposed males were within the historical control range. For males at all dietary concentrations, statistically significant higher levels of potassium were observed compared to control, however, the majority of individual values were within the historical control range.

 

Although macroscopic necropsy findings at termination did not indicate any obvious effect of dietary exposure to the test material at 3000, 7500 and 18000/12000 ppm, a number of organ weights attained statistical significance compared to control: Males at the 18000/12000 ppm dietary level showed lower absolute liver weight and higher relative liver weights compared to control, although the majority of individual values were within the historical control range. Microscopic evaluation revealed minimal diffuse atrophy of hepatocytes for both sexes at 18000/12000 ppm and, to a lesser extent, at 7500 ppm.

 

Females at the 7500 and 18000/12000 ppm dietary levels, showed lower absolute and relative pituitary weight compared to control, although the majority of individual values were within the historical control range. Microscopic evaluation revealed minimal diffuse atrophy of pituicytes for females at the 18000/12000 ppm and, to a lesser extent, at the 7500 ppm dietary level. Males receiving diets containing 7500 ppm or 18000/12000 ppm of the test item showed lower absolute and relative seminal vesicles weights compared to control. At the 18000/12000 ppm exposure level a number of individual seminal vesicle values were outside the historical control range and microscopic evaluation revealed a minimal or mild reduction in secretion in the seminal vesicles. This finding at the high dietary exposure level was therefore considered to be treatment related. For males receiving 7500 ppm of the test material all individual seminal vesicles weights were within the historical control and in the absence of any supporting histopathological change, the decrease in seminal vesicle weight at this dietary concentration was considered not to be of toxicological significance. Females at all dietary levels showed lower absolute and body weight relative spleen weights compared with control but all individual absolute and body weight relative values for treated animals were within the historical control range. Microscopic evaluation revealed lower extramedullary haematopoiesis for females at the 7500 or 18000/12000 ppm dietary levels compared to control, therefore the lower spleen weights at these exposure levels were considered to be related to treatment. At the 3000 ppm exposure level, there was no evidence of histopathological change and the lower spleen weights were considered to be of no toxicological significance.

 

These differences in organ weight and accompanying microscopic changes were considered to reflect comparative differences in food consumption and body weight gain and, in the zona glomerulosa, the effects of dehydration on electrolyte imbalance, (Levin et al., 1993). Other microscopic changes also considered to be attributable to these differences included: Minimal or mild diffuse hypertrophy of the zona glomerulosa of the adrenal cortex in both sexes at the 7500 or 18000/12000 ppm dietary levels and minimal diffuse atrophy of the zona fasciculata of the adrenal cortex in both sexes at the 18000/12000 ppm dietary level and, to a lesser extent, males at the 7500 ppm dietary level.

 

Minimal or mild increase in adipose tissue was observed in the bone marrow samples of males at the 7500 ppm dietary level and in both sexes at the 18000/12000 ppm dietary level. Minimal reduced secretion in the prostate gland of one male was observed at the 18000/12000 ppm dietary level. Minimal or mild decreased cortical lymphocytes of the thymus was observed in females at the 7500 ppm dietary level and in both sexes at 18000/12000 ppm dietary level.

 

Other microscopic changes, considered to be more serious in nature, were also observed. For the kidney, mild or moderate foci of basophilic tubules were apparent for a few animals at the 18000/12000 ppm dietary level, these animals also showed minimal or mild single cell death in the affected tubules. Minimal to moderate pelvic dilatation was also apparent for males at the 7500 ppm dietary level and both sexes at the 18000/12000 ppm dietary level and mild hypertrophy of the collecting ducts in the papilla was seen a few animals at the 18000/ 12000 ppm dietary level. For the urinary bladder, minimal to moderate diffuse hyperplasia of the urothelium, with associated minimal to moderate chronic active inflammation, was apparent for females at the 7500 ppm dietary level and in both sexes at the 18000/12000 ppm dietary level. At the highest exposure level, a few affected animals also showed erosion or ulceration of the mucosa and one female showed minimal focal submucosal haemorrhage. Two females at the 7500 ppm dietary level also showed minimal inflammatory cell infiltration of the submucosa. The observed changes in the urinary bladder and kidneys were considered adverse and due to an irritant effect of the test item, or a metabolite, to the urothelium. The more severe effects apparent in the urinary bladder was probably due to the longer time that the urinary bladder mucosa was exposed to the test item or metabolite. Microscopic changes were also apparent in the mesenteric lymph nodes, with minimal to moderate intrasinusal congestion/erythrophagocytosis being observed in males at the 7500 ppm dietary level and in both sexes at the 18000/12000 ppm dietary level.

 

Mating performance, pregnancy rate, and gestation length were unaffected by dietary exposure to the test material. At the 18000/12000 ppm dietary concentration, the mean number of corpora lutea was lower than control, resulting in a lower number of implantations and litter size on post-natal days 1 and 4. However, no obvious effect on pre-natal and post-natal survival or sex ratio was observed at this exposure level. At the 3000 and 7500 ppm dietary concentrations, there were no obvious effects on numbers of corpora lutea, implantations, pre-and post implantation losses, litter size or sex ratio at birth/postnatal day 1 and post-natal day 4 of age or on post-natal pup survival.

 

At the 18000/12000 ppm dietary concentration, offspring bodyweight weight and litter weights on post-natal days 1 and 4 and offspring body weight gain to post-natal day 4 were lower than control. Offspring performance during assessment of surface righting on post-natal day 1 was also inferior to control at the highest exposure level. At the 7500 ppm dietary concentration, offspring bodyweight weight on post-natal days 1 and 4, offspring body weight gain to post-natal day 4 and litter weight on post-natal day 4 were lower than control. Offspring performance during assessment of surface righting was also inferior to control at the intermediate exposure level. At the 3000 ppm dietary concentration, offspring body weights, litter weights, offspring body weight gain and surface righting ability of the offspring were considered to be unaffected by maternal dietary exposure to the test material.

 

At the 7500 and 18000/12000 ppm dietary concentrations, there was a slightly higher incidence of clinical signs among offspring but these were concentrated in only a few litters (no milk visible in stomach (2 and 3 litters at 7500 ppm and 18000/12000 ppm respectively), cold (1 litter at 18000/12000 ppm), weak (1 litter at 18000/12000 ppm), small (2 and 3 litters at 7500 ppm and 18000/12000 ppm respectively) and distended abdomen (1 litter at 7500 ppm)), and did not indicate a clear effect of maternal dietary exposure on offspring development.

 

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.

 

Oral administration of the test material resulted in treatment related effects in animals of both sexes treated with 18000/15000 ppm and in males treated with 7500 ppm.

 

There were no clinical signs evident for animals of either sex treated with 3000, 7500 or 18000/15000 ppm. The physical condition of males treated with 18000 ppm was affected with reductions in body weight development during Weeks 1 and 5 of treatment, although body weight remained within 7% of control values throughout the duration of the study. Subsequently a reduction in overall body weight gain was evident in these males. A reduction in overall food consumption was evident in males at this dosage and a reduction in food conversion efficiency during Week 1 was evident. Females treated with 18000/15000 ppm showed a reduction in body weight gain and food consumption during the final two weeks of gestation and during lactation.

 

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.

Effects on developmental toxicity

Description of key information

One key developmental toxicity study (OECD 414) was identified for Resin acids and Rosin acids, fumarated esters with glycerol.

1) Resin acids and Rosin acids, fumarated esters with glycerol (CAS# 97489-11-7): 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 range-finding developmental toxicity study (Envigo Research Laboratories, 2017), there was no effect of maternal treatment with Resin acids and rosin acids, fumarated esters with glycerol 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 developmental toxicity NOAEL in rats was therefore determined to be 15000 ppm (equivalent to 1198.5 mg/Kg bw/day).

Three key screening reproductive/developmental toxicity studies were identified for Resin acids and rosin acids, maleated, esters with pentaerythritol; Resin acids and rosin acids, fumarated, esters with glycerol; andResin acids and rosin acids, fumarated, esters with pentaerythritol.

1) Resin acids and rosin acids, maleated, esters with pentaerythritol, CAS# 94581-17-6): The No Observed Adverse Effect Level (NOAEL) for survival, growth and development of the offspring was considered to be 3000 ppm, based on effects on body weight, body weight gain and surface righting ability at 7500 ppm and 18000/12000 ppm.

2) Resin acids and rosin acids, fumarated, esters with glycerol (CAS# 97489-11-7): The No Observed Adverse Effect Level (NOAEL) for developmental toxicity was determined to be 7500 ppm, based on adverse treatment-related effects on offspring body weight observed at the highest concentration tested.

3) Resin acids and rosin acids, fumarated, esters with pentaerythritol (CAS# 94581-15-4): The ‘No Observed Effect Level’ (NOEL) for reproductive/developmental toxicity was considered to be 1000 mg/Kg/day since no treatment-related effects were adverse effects were observed at the highest dose tested.

Supporting data is available 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).

For Rosin, fumarated (CAS# 65997-04-8), there was no clear evidence of test substance-related effects on reproduction of F0 males and females or on survival and development of F1 pups. Therefore, the NOAEL for reproductive parameters was considered to be 10000 ppm (males 651-889 mg/Kg bw/day; females 449-995 mg/Kg bw/day), the highest concentration tested.

 

For Resin acids and rosin acids, esters with pentaerythritol (CAS# 8050-26-8), the no observed adverse effect level (NOAEL) for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 20000 ppm (the highest concentration tested) for males and females (equivalent to received doses of 1864 mg/Kg bw/day and 1757-2054 mg/Kg bw/day, respectively).  

 

For Rosin (CAS# 8050-09-7), the NOAEL for reproductive/developmental toxicity in Sprague-Dawley rats was considered to be 3000 ppm (equivalent to 248 mg/Kg bw/day in males and 309 mg/Kg bw/day in females), based on slightly reduced mean litter and pup weights when compared to controls in dams exposed at 10000 ppm.

Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

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.

 

Clinical signs, body weight change, food and water consumptions were monitored during the study. All females were terminated on gestation day 20 and subjected to gross necropsy including examination of the uterine contents. The number of corpora lutea, number, position and type of implantation, placental weights, foetal weight, sex and external and internal macroscopic appearance were recorded. Half of the pups from each litter were examined for detailed skeletal development and the remainder were subjected to detailed visceral examination.

 

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/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).

Justification for classification or non-classification

Not classified for reproductive or developmental toxicity according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 or UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS).