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Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Two key Guideline (OECD 422) studies were identified for Rosin, fumarated and Rosin, maleated.

1) Rosin, maleated (CAS# 8050-28-0): The NOAEL and NOEL (No Observed Effect Level) for reproductive toxicity were determined to be 3500 ppm (equivalent to mean achieved doses of 224.9 mg/Kg bw/day (pre-pairing); 245.8 mg/Kg bw/day (Gestation); and 371.9 mg/Kg bw/day (Lactation)).

2) Rosin, fumarated (CAS# 65997 -04 -8): The NOAEL for reproductive parameters was considered to be 10,000 ppm (equivalent to 651-889 mg/Kg bw/d in males and 449-995 mg/Kg bw/d in females).

Supporting data from two screening studies (OECD 421) is also available for Rosin (the precursor of all substances included in this category) and Resin acids and rosin acids, esters with pentaerythritol (a Rosin reaction product).

1) Rosin (CAS# 8050-09-7): The NOAEL for reproductive 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), based on a slight decrease in the mean number of implantation sites resulting in a subsequent slight reduction in litter size at the highest concentration tested.

2) Resin acids and rosin acids, esters with pentaerythritol (CAS# 8050-26-8): The NOAEL for reproductive 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), based on a lack of observed treatment-related effects on reproductive performance of the parental females or on survival and development of the F1 pups.

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

Information exists to characterise the reproductive toxicity of Rosin Adducts and Rosin Adducts Salts. These are formed when rosin reacts with maleic anhydride (or maleic acid) or fumaric acid yielding a maleated rosin adduct or a fumarated rosin adduct, respectively; the rosin adduct salts are simply the rosin adducts that have been reacted with an appropriate base. The available data includes results obtained from testing Rosin, fumarated; and Rosin, maleated along with supporting data for 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 Guideline (OECD 422) combined repeated dose, reproductive/developmental toxicity screening study (Harlan Laboratories Ltd., 2015a), the test material (Rosin, maleated; CAS# 8050-28-0) was administered daily in dietary mixtures at concentrations of 0, 1750, 3500 or 7000 ppm to male rats for 43 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 post partum.

 

Treatment resulted in slightly higher test item intake in females compared to males during the pre-pairing period. Intake remained fairly constant throughout the entire study, except during the lactation phase when, due to an increase in food consumption, intake in females was almost two-fold higher than during the preceding gestation period.

 

Adverse toxic effects were observed in females in group 4 (7000 ppm) during the pre-pairing period. A severe reduction in food consumption resulting in body weight loss, significantly reduced body weights and significantly reduced food conversion efficiency were recorded. Further, clinical signs indicating worsening of the animal condition like stiff gate, hunched posture and ruffled fur were observed in most females in this group between days 12 and 14 of the pre-pairing period. Four females were found dead between day 12 and 14 and remaining females were terminated for ethical reasons on day 14 of the pre-pairing period. During necropsy, reddish discoloration of the cecum was observed in one female. This finding was histologically correlated with moderate submucosal edema/congestion. In one further female, pale discoloration of kidney and pelvic dilation was observed during the necropsy and histopathological examination confirmed kidney as a target organ in the females in the high concentration group. The pathogenesis of these changes was uncertain, however an acute/subacute impairment of the urinary outflow was suspected. The kidney findings did not appear to be linked to the cause of death as its grade severity was higher in animals killed in extremis when compared to animals found dead. Starvation caused by the aversion to test item containing food possibly contributed to the effects on body weights. However, there was no clear evidence that the food aversion was the immediate reason of the severity of the effects and bad condition of females. Thus, the reason for the mortality of females in the high-dose group remained unclear.

 

Treatment-related reduction in food consumption, food conversion efficiency, water consumption, body weight gain and body weights were also observed in males in the high concentration group (7000 ppm), however, less severe than in females. Although food and water consumption remained also slightly lower during the entire study, they recovered after the significant reduction observed at the beginning of the treatment. A body weight loss was observed at the beginning of the treatment followed by a reduced body weight gain. This resulted in reduced body weights observed until the end of the study.

 

During necropsy of males in the high concentration group, reduced adrenal weights were recorded but no other macroscopic findings. Histopathological examination revealed minimal decreased lymphocytes in the spleen and mesenteric lymph nodes which was considered to be a result of a non-specific stress.

 

In group 3 (3500 ppm), a slight and reversible reduction in food consumption and food conversion efficiency was observed in males and females. This resulted in a slight and reversible reduction in body weights, however without an effect on absolute body weights. A slight reduction in water consumption was recorded in males. During necropsy, reduced adrenal weights were recorded in females. Histopathological examination revealed no test item-related findings in any gender. No further test item-related effects were observed in any group. No test item-related findings were recorded in group 2 (1750 ppm).

 

Due to early termination of females in group 4, reproduction and developmental parameters were not assessed in this group. Mating performance, fertility, duration of gestation, number of corpora lutea, number of implantation sites and post implantation loss, were not affected by the treatment with the test item at any dose level. In group 3 (3500 ppm), reduced pup body weights and reduced pup body weight gain were recorded from day 1 to 4 of the lactation period. The differences of mean values were statistically significant if compared to the respective current control values and the values in group 3 were slightly below the historical control range. Therefore, this effect was considered to be related to the treatment with the test item. The changes in pup body weights were concomitant with slight reduction in maternal weight and weight gain. In group 2, pup body weights and body weight gain were considered not to be affected by the treatment with the test item. Further, an increased postnatal loss was observed in both dose-groups. However, the relation of this observation to the treatment with the test item was not conclusively established.

 

Based on the results of this study, a NOAEL (No Observed Adverse Effect Level) for systemic toxicity was established at the dose level of 3500 ppm.

 

The NOAEL and NOEL (No Observed Effect Level) for reproduction were set at the dose level of 3500 ppm whereas the NOEL and NOAEL for developmental toxicity was established at the dose level of 1750 ppm due to the reduced body weights and body weight gain in pups at the dose level of 3500 ppm.

 

In another key Guideline (OECD 422) combined repeated dose, reproductive/developmental toxicity study (Inveresk Research 2004a), the test material (Rosin, fumarated; CAS# 65997-04-8) 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.

 

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).

Results from two supporting screening studies are presented below:

In a Guideline (OECD 421) reproductive/developmental toxicity screening study (Inveresk Research, 2003a), 10 rats/sex/group were exposed to the test material (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) and the NOAEL for subchronic toxicity was considered to be 1000 ppm in males (equivalent to 84 mg/Kg bw/d) and 3000 ppm in females (equivalent to 309 mg/Kg bw/d) based upon reduced feed consumption and lower weight gain in animals consuming higher dietary concentrations of the test material.

 

In another Guideline (OECD 421) reproductive/developmental toxicity screening study (Inveresk Research, 2004b), 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.  

Effects on developmental toxicity

Description of key information

Two key Guideline (OECD 414) developmental toxicity studies and two Guideline (OECD 422) reproductive/developmental toxicity studies were identified for Rosin, fumarated; and Rosin, maleated.

OECD 414

 

1) Rosin, fumarated (CAS# 65997-04 -8): 10000 ppm (equivalent to a mean achieved dose of approximately 908.2 mg/Kg bw/day) was determined to be the maternal No Observed Adverse Effect Level (NOAEL) for pregnant females. There was no evidence that dietary exposure to the test material at 10000 ppm increased the occurrence of teratogenic effects. This dose was therefore considered to be the NOAEL for developmental toxicity.

2) Rosin, maleated (CAS# 8050-28-0): The NOEL for the developing conceptus was considered to be a maternal dietary exposure to 500 ppm of the test material. The NOAEL for developmental toxicity was also considered to be at least 500 ppm of the test material (equivalent to a mean achieved dose of 41.2 mg/Kg bw/day), based on effects on foetal and placental weights and numerous skeletal findings indicating incomplete ossification or no ossification for many regions of the skeleton at the 1500 ppm level.

 

OECD 422

 

1) Rosin, maleated (CAS# 8050-28-0): The NOAEL and NOEL (No Observed Effect Level) for developmental toxicity was established at the dose level of 1750 ppm, based on reduced body weights and body weight gain in pups at the dose level of 3500 ppm.

 

2) Rosin, fumarated (CAS# 65997-04-8): The NOAEL for developmental toxicity was considered to be 10000 ppm (449-995 mg/Kg bw/d), the highest concentration tested.

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

Information exists to characterise the developmental toxicity of Rosin Adducts and Rosin Adducts Salts. These are formed when rosin reacts with maleic anhydride (or maleic acid) or fumaric acid yielding a maleated rosin adduct or a fumarated rosin adduct, respectively; the rosin adduct salts are simply the rosin adducts that have been reacted with an appropriate base. The available data includes results obtained from testing Rosin, fumarated; Rosin, maleated; and Resin acids and Rosin acids, maleated, sodium salts and is summarised below.

In a key Guideline (OECD 414) pre-natal developmental toxicity study (Envigo CRS, S.A.U, 2018a), the test material (Rosin, fumarated; CAS# 65997-04-8) was administered by continuous dietary admixture to three groups of Sprague-Dawley rats, between gestation days 3 and 19 (inclusive) at dietary concentrations of 2500, 5000, and 10000 ppm (equivalent to mean achieved dosages of approximately 0, 188.2, 369.3 and 908.2 mg/Kg bw/day). An additional group of female rats was treated with basal laboratory diet to serve as a control.

 

Clinical signs, body weight change, and food consumption was 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 weights, sex and external and internal macroscopic appearances were recorded. Half of the pups of each litter were examined for detailed skeletal development and the remainder were subjected to detailed visceral examination.

 

No mortality and no clinical signs were apparent for adult females throughout the study at dietary concentrations of 2500, 5000 or 10000 ppm of the test material. Hunched posture was transiently observed in a few of the animals exposed to the test material at 5000 or 10000 ppm, but was not considered to be a toxic effect since the incidence was unrelated to exposure concentration.

 

Food consumption in animals exposed at 10000 ppm was higher than the control group during the exposure period. Despite there being no effect on absolute

body weight, a trend to lower body weight gain was observed in the animals exposed at 10000 ppm, especially between days 4 and 10 of gestation. However, this trend was not considered toxicologically relevant since it was a transient effect and no relevant differences were observed in body weight gain later during gestation, nor were there any effects on the general well-being of the dams in this group.

 

There were no relevant findings in the observations derived from hysterectomy. However, slightly lower mean foetal body weights at the10000 ppm exposure

level (compared to control) could help explain the slightly lower placenta and uterus weights observed at this exposure level, as was evident when mean placental or uterus weights were compared to foetal weights. There was no effect on the sex ratio post exposure to the test material.

 

Gross necropsy did not reveal any remarkable findings.

 

A low incidence of major foetal abnormalities was observed at 10000, 5000 and 2500 ppm and in the control group (1/312 in Control, 1/263 at 2500 ppm, 2/294

at 5000 ppm and 2/341 at 10000 ppm). These were considered to be incidental, given their incidence and distribution across the treatment and control groups.

 

Skeletal examination of the foetuses showed a slightly increased incidence of delayed/incomplete ossification/unossified sternebrae in the high and low exposure groups (9% foetuses in Control, 21% at 2500 ppm, 10% at 5000 ppm and 21% at 10000 ppm) and short supernumerary 14thribs (8% of foetuses in Control, 20% at 2500, 8% at 5000 and 22% at 10000 ppm). However, the incidences in the high and low exposure groups were similar. Furthermore, the incidence of delayed/incompletely ossification/unossified sternebrae at 10000 and 2500 ppm was within the Historical Control range. Given the distribution of these findings within the treatment groups, they were not considered adverse or treatment-related.

 

Skeletal and visceral examination of the foetuses showed the presence of major abnormalities in one foetus at 10000 ppm (litter no. 105, foetus no. 2), two at

5000 ppm (litter no. 65, foetus no. 14 and litter no. 75, foetus no. 8), one at 2500 ppm (litter no. 38, foetus no. 2) and one in the control group (litter no. 18, foetus no. 6). Given their incidence and distribution, these abnormalities were considered unrelated to treatment with the test material.

 

Based on the results of this study, 10000 ppm was determined to be the maternal No Observed Adverse Effect Level (NOAEL) for pregnant females. Although a statistically significant effect on body weight gain between days 4 and 10 was observed at 10000 ppm, it was considered non-adverse, since it was a transient effect and no relevant differences were observed in body weight gain later during gestation, nor were there any effects on the general well-being of the dams in this group.

 

With respect to effects on embryofoetal development, 10000 ppm was determined to be the NOAEL. Although slight but statistically significant reduced foetal

body weights were observed, this was not considered an adverse effect, due to the small magnitude of the change and because the mean value was within the

Historical Control range. Despite some abnormalities in all dietary exposure groups as well as in control foetuses, there was no evidence that dietary exposure to the test material at 10000 ppm increased the occurrence of teratogenic effects. This dose was therefore considered to be the NOAEL.

In another key Guideline (OECD 414), pre-natal developmental toxicity study (Envigo Research Laboratories, 2017b), the test material (Rosin, maleated; CAS# 8050-28-0) 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 500, 1500, and 3000 ppm equivalent to mean achieved dosages of approximately 41.2, 122.9 and 217.4 mg/kg bw/day. A further group of twenty-four time mated females was treated with 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 weights, sex and external and internal macroscopic appearances were recorded. Half of the pups of each litter were examined for detailed skeletal development and the remainder were subjected to detailed visceral examination.

 

No mortality was and no clinical signs were apparent for adult females throughout the study at dietary concentrations of 500, 1500 or 3000 ppm of the test

material.

 

At a dietary concentration of 3000 ppm of the test material, marginal mean body weight loss was apparent during the first day of dietary exposure and subsequent

bodyweight gains were often statistically significantly lower than control animals throughout gestation. Cumulative body weight gain from the start of treatment was statistically significantly lower than control animals from gestation day 5 and overall body weight gain, when adjusted for the contribution of the gravid uterus, remained statistically significantly lower than control. In rats exposed to 1500 ppm of the test material, statistically significantly lower body weight gain

was apparent during gestation days 17 to 20. Prior to this, no statistically significant difference in body weight gain had been apparent and there were no statistically significant differences from control animals apparent for cumulative body weight gain, including overall body weight gain when adjusted for the contribution of the gravid uterus. No statistically significant differences in body weight gain during gestation were apparent for females that received 500 ppm of the test material. Food consumption throughout gestation was statistically significantly lower in rats exposed to the test material at 3000 ppm when compared with control animals. No obvious effects on food consumption during gestation were observed in animals exposed to the test material at 500 or 1500 ppm.

 

Gross necropsy on Gestation Day 20, did not reveal any remarkable findings (macroscopic abnormalities) in adult animals in any of the treatment groups.

 

There was no effect of maternal dietary exposure 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 at dietary exposures of 500, 1500 or 3000 ppm of the test material.

 

Maternal dietary exposure to 3000 ppm of the test material resulted in statistically significantly lower mean foetal, litter and placental weights when compared to

the control animals. At the 1500 ppm maternal dietary exposure level, mean foetal and placental weights were statistically significantly lower than the control group. Litter weight and total placental weight were also lower when compared to the control group, but differences failed to attain statistical significance. No

statistically significant differences in mean foetal and placental weights were observed in the 500 ppm treatment group when compared to the control animals.

 

External examination at necropsy revealed a higher incidence of visually small foetuses in the 3000 ppm and, to a lesser extent, the 1500 ppm maternal dietary exposure groups, when compared to the control. No such effects were apparent in the 500 ppm treatment group. Neither the type, incidence nor the

distribution of visceral findings observed in the foetuses indicated any effect of maternal dietary exposure to 500, 1500 or 3000 ppm of the test material.

 

Maternal dietary exposure to 3000 ppm of the test material, resulted in a clear effect on a large number of ossification parameters affecting most regions of the

skeleton. There was (generally) an increase in both the number of foetuses and litters affected compared to the control group and statistically significant differences from control in foetal incidence for these parameters. At the 1500 ppm maternal dietary exposure level, there was a similar effect on ossification parameters, although changes were not quite as extensive and generally occurred at a lower incidence than that observed at the 3000 ppm dietary exposure level. Again, there was generally an increase in both the number of foetuses and litters affected compared to the control group and statistically significant differences from control in foetal incidence for these parameters. Neither the type, incidence nor the distribution of skeletal findings observed for foetuses indicated any effect of maternal dietary exposure to 500 ppm of the test item.

 

Based on the results of this study, dietary exposure to 500 ppm of the test material (equivalent to an achieved dosage of approximately 41.2 mg/kg bw/day of the

test material) was considered to be a clear No Observed Effect Level (NOEL) for the pregnant dam. The No Observed Adverse Effect Level (NOAEL) for the pregnant dam was considered to be a dietary exposure to 1500 ppm of the test item (equivalent to an achieved dosage of approximately 122.9 mg/kg bw/day of the test material), due to effects on body weight gain and food consumption apparent at a dietary level of 3000 ppm.

 

The NOEL for the developing conceptus was considered to be a maternal dietary exposure to 500 ppm of the test item. The NOAEL for developmental toxicity

was also considered to be at least 500 ppm of the test material, due to effects on foetal and placental weights and numerous skeletal findings indicating incomplete ossification or no ossification for many regions of the skeleton at the 1500 ppm level.

 

In a key Guideline (OECD 422), reproductive/developmental toxicity study (Harlan Laboratories Ltd, 2015a), the test material (Rosin, Maleated; CAS# 8050-28-0) was administered daily in dietary mixtures at concentrations of 0, 1750, 3500 or 7000 ppm to male rats for 43 days and to female rats for 14 days prior to pairing,

through the pairing and gestation periods until the F1 generation reached day 4 post partum. Treatment with the test item resulted in stiff gait, hunched posture and ruffled fur in females in group 4 at the end of the pre-pairing period. In group 4, four females were found dead during the pre-pairing period. Remaining females in this group were terminated on day 14 of the prepairing period due to significant reduction in food consumption, body weight loss and clinical signs indicating bad condition. No further test item-related findings were noted. Functional observational battery and locomotor activity were similar in all groups in males and females. In males, treatment with the test item caused a dosedependent but reversible and therefore not adverse reduction in food consumption in groups 3 and 4. In females, treatment with the test item caused adverse reduction in food consumption in group 4 and a reversible and therefore not adverse reduction in food consumption in group 3. A dose dependent reduction in water consumption was recorded only in males of groups 3 and 4. The treatment with the test item caused a dose-dependent but reversible and therefore not adverse reduction in body weight gain in males of groups 3 and 4. In females, body weight loss and adverse reduction in body weights in group 4 was noted. In group 3, reversible and therefore not adverse reduction in body weight gain without significant changes in body weights were observed. The hematology or biochemistry parameters were not different from the control group.

 

Mating performance, fertility, duration of gestation, number of corpora lutea, number of implantation sites and post implantation loss were not affected by the

treatment with the test item at any dose level. An increased postnatal loss was observed in groups 2 and 3 but this effect was not clearly correlated with the dose levels. Regarding the organ weights, a decrease in adrenal weights in males in group 4 and females in group 3 was recorded. In males, no test item-related findings were recorded during the necropsy. During histopathological examination, minimal decreased lymphocytes in the spleen and mesenteric lymph nodes were observed in this gender in group 4. This finding was considered to be related to a non-specific stress. In females, test item-related findings were observed only in group 4 animals found dead or prematurely killed. During necropsy, a reddish discoloration of caecum was found in one female, pale discoloration of kidney and pelvic dilation were noted for another one. Histopathological findings consisted of kidney changes: pelvis urothelium hyperplasia and luminal pelvis dilatation associated with tubular dilatation/degeneration/regeneration. Further, decreased lymphocytes in lymphoid organs were found in some descendents, which was considered to be related to a nonspecific stress. The reddish discoloration of the cecum observed at necropsy was correlated histologically with moderate submucosal edema/congestion.

 

Treatment with the test item did not induce any pathology findings in the reproductive organs of males or females in any group. No test item-related findings were observed in pups at first litter check or during lactation. Righting reflex and sex ratios were unaffected. The test item resulted in reduced pup body weights and reduced pup body weight gain during lactation in group 3. Pup body weights and body weight gain in group 2 were considered not to be affected by the treatment with the test item. No test item-related findings were noted during necropsy of pups in any group.

 

Based on the results of this study, a NOAEL (No Observed Adverse Effect Level) for general toxicity was established at the dose level of 3500 ppm. The NOAEL and NOEL (No Observed Effect Level) for reproduction were set at the dose level of 3500 ppm whereas the NOEL and NOAEL for development was established at the dose level of 1750 ppm due to the reduced body weights and body weight gain in pups at the dose level of 3500 ppm.

 

In another Guideline (OECD 422), reproductive/developmental toxicity study (Inveresk Research, 2004a), the test material (Rosin, fumarated; CAS# 65997-04-8) was administered in the diet to female rats at concentrations of 0, 1000 ppm (79-108 mg/Kg bw/d), 3000 ppm (196-292 mg/Kg bw/d), and 10,000 ppm (449-995 mg/Kg bw/d) throughout pregnancy until termination on Day 4 of lactation. Food consumption and mean body weights were decreased at 10,000 ppm and 3000 ppm, with high dose animals also showing an increase in total bilirubin and decreased adrenal weight.  The lower of these two values will be used as the

maternal (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 litter 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 developmental effects was considered to be 10,000 ppm (449-995 mg/Kg bw/d).

In a supporting dose range-finding developmental toxicity study (Charles River Laboratories, 2017), the test material (UVCB Rosin maleated (10 % MA w/w), CAS# 8050-28-0)), was administered ad libitum by dietary admixture at concentrations of 500, 1500 and 3000 ppm (achieved doses of 44, 123 and 284 mg/Kg body

weight/day, respectively) to groups of 6 mated Sprague Dawley rats from gestation days 3 to 20. A control group received diet from the same batch as treated animals and under the same conditions.

 

No test material-related mortality and no signs of exposure-related clinical toxicity were observed through the study period. Treatment with the test material was associated with slight maternal toxicity in the high dietary exposure group (reduced body weight gain and food consumption) principally towards the end of the gestation period. Consistent with the maternal response at 3000 ppm, lower mean foetal weight and slightly delayed ossification of several bones (principally of the paws, sternum and caudal vertebrae) were noted.

 

Slightly delayed ossification was also noted for the caudal vertebra only in the lower dose groups with evidence of a dose-response relationship in the number

of affected foetuses and litters in the absence of any test material-related effect on mean foetal weight. Although probably test material-related, this minor finding represents normal variation that exists in the rate of development of some skeletal elements in a control population. As there was no structural change in the bones to compromise foetal wellbeing and no other test item-related effects on embryo-foetal development, the slightly higher incidence of this minor and isolated skeletal variation was considered to be of no toxicological significance.

 

Based on the results observed, the maternal systemic toxicity NOAEL was determined to be 1500 ppm (equivalent to an achieved dosage of 123 mg/Kg bw/day) due to effects on body weight gain and food consumption at the dietary level of 3000 ppm (equivalent to an achieved dosage of 284 mg/Kg bw/day). The developmental toxicity NOAEL was determined to be 1500 ppm (equivalent to an achieved dosage of 123 mg/Kg bw/day) due to an effect on foetal weight and

delayed ossification of a several regions of the skeleton at the dietary level of 3000 ppm (equivalent to an achieved dosage of 284 mg/Kg bw/day).

In another supporting dose range-finding developmental toxicity study (Charles River Laboratories, 2018), the test material (UVCB Resin acids and Rosin acids, maleated, sodium salts, 15% MA, w/w, (CAS no. 68201-60-5)), was administered ad libitum by dietary admixture at nominal concentrations of 500, 1500 and 3000 ppm (equivalent to average test material intake (period of exposure - GD’s 3 to 20) of 39, 120 and 250 mg/Kg body weight/day, respectively) to groups of 6 mated Sprague Dawley rats from gestation days 3 to 20. A control group received diet from the same batch as treated animals and under the same conditions.

 

Maternal parameters monitored included morbidity/mortality, clinical signs, body weight andfood consumption. The females were submitted to a caesarean examination on gestation Day 20 and litter parameters were recorded. At necropsy, the females were examined macroscopically and thegravid uterus was weighed. All foetuses were weighed, sexed and examined for external abnormalities. All foetuses were then processed for skeletal examination.

 

No test material-related mortality and no signs of exposure-related clinical toxicity were observed through the study period. There was no effect on mean body weight gain or mean terminal body weight in any group. Food consumption for treated animals was comparable with or superior to that of the control group. Gross necropsy did not reveal any remarkable findings. One female (no. 7) fed diet containing 500 ppm exhibited renal pelvis dilation but this isolated finding in the low dietary exposure group was considered incidental. There was no effect on gravid uterus weight in any group and all 6 females in each group were pregnant with viable foetuses at the terminal caesarean examinations. There was no influence of dietary exposure (initiated on gestation Day 3) in any group on the pre-implantation data (mean numbers of corpora lutea and implantations and the corresponding percentage pre-implantation loss) and embryo-foetal survival was unaffected by exposure to the test material.

 

There was no effect on mean foetal weight or sex ratio in any group. External examination of the foetuses revealed one control group foetus (no. 2) from dam no. 5 with brachydactyly of the rightforepaw and gastroschisis. These malformations observed in a single control group foetus were considered incidental and no external abnormalities were observed in the treated groups. Skeletal examinations showed that the same control group foetus also showed absence of phalanx of the right forepaw, split palatine and absence of metatarsal with absence of phalanx of the right hindpaw. These malformations observed in a single control group foetus in were considered incidental. Amongst the other less severe skeletal anomalies and variations, there was an exposure related increased incidence of foetuses/litters with unossified metacarpal of 5th digit in the 500, 1500 and 3000 ppm groups, respectively when compared with the control group. This anomaly was commonly observed in the historical control data (HCD) of the laboratory and the foetal incidence remained within the HCD range and was therefore considered incidental. The incidences of the other skeletal anomalies and variations were comparable with the concurrent control and/or the historical control data or there was no evidence of an expsoure response relationship in the number of affected foetuses and/or litters to suggest any association with oral dietary exposure.

 

Oral administration (dietary admixture) of the test material (UVCB Resin acids and Rosin acids, maleated, sodium salts, 15% MA w/w, (CAS no. 68201-60-5)) to pregnant Sprague Dawley rats during gestation (Days 3 to 20) at nominal concentrations of 500, 1500, 3000 ppm was not associated with any maternal toxicity or effect on early implantation, embryo-foetal survival or skeletal development. Based on the results observed, the maternal systemic toxicity NOAEL and the developmental toxicity NOAEL were determined to be 3000 ppm (equivalent to average test material intake (period of exposure - GD’s 3 to 20) of 250 mg/Kg body weight/day, respectively)

Justification for classification or non-classification

Rosin Adducts and Rosin Adducts Salts are 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).

 

A detailed description of the available physicochemical, environmental and toxicological data of the Category 3 substances is presented at the beginning of the Testing Strategy Document attached to Section 13 of the dossier. The physico-chemical data indicate that there are similarities between Resin acids and Rosin acids, maleated, sodium salts (68201-60-5); Resin acids and Rosin acids, maleated, potassium salts (85409-27-4); and Resin acids and Rosin acids, maleated, calcium salts (91722-01-9) to Rosin fumarated (65997-04-8) (provided in Table 3 of the Category Justification Document attached to Section 13 of the dossier). No data are available for the mammalian toxicity of Resin acids and Rosin acids, maleated, sodium salts or Resin acids or Rosin acids, maleated, calcium salts.

 

In order to provide supportive evidence regarding the structural similarity of Resin acids and Rosin acids, maleated, sodium salts; Resin acids and Rosin acids, maleated, potassium salts; and Resin acids and Rosin acids, maleated, calcium salts to Rosin fumarated, an analytical assessment was performed (Kemira (2017) Lab Report: Isolation and analysis of rosin from rosin soap).

 

The comparison of IR spectra (Figure 1 of Isolation and analysis of rosin fromrosin soap (Kemira (2017) Lab Report: Isolation and analysis of rosin from rosin soap) clearly shows that the rosin extracted from rosin soap is similar to rosin-fumarated in the relevant range from 1600-1800 cm-1. This range is characteristic for acid and anhydride groups. Rosin maleated shows different spectral-bands in this spectral region, due to the presence of the anhydride group. IR analysis of the rosin soap (50% solution and dried) confirms the absence of the characteristic anhydride band of maleated rosin. This analysis shows that the anhydride group of rosin maleated is hydrolysed to the corresponding tri-carboxylic-acid in the rosin-soap production process. This carboxylic acid is chemically very similar to fumaropimaric acid (stereo-isomers). 

 

In conclusion, the results of these analyses demonstrate that the substances Resin acids and Rosin acids, maleated, sodium salts; Resin acids and Rosin acids, maleated, potassium salts; and Resin acids and Rosin acids, maleated, calcium salts are more structurally similar to Rosin fumarated than to Rosin maleated. In addition, the physico-chemical data also indicate that there are similarities between the Resin acids and Rosin acids, maleated, sodium; calcium; and potassium salts, and Rosin fumarated.  Therefore, Rosin fumarated is considered the most appropriate Category 3 substance for read across to Resin acids and Rosin acids, maleated, sodium, potassium, and calcium salts.