Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

Endpoint summary

Description of key information

Effects dominated by content of DEA: Target organs are the haemeopoietic system (mild anemia) and the kidneys.

Calculated NOAELs / LOAELs based on DEA content of 5.5%:

oral: LOAEL = 255mg/kg (90 -day study), NOAEL = 236 (EOGRTS)

Inhalation NOAEC = 0.27mg/L

dermal LOAEL = 582mg/kg

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

236 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEC

272 mg/m³

Study duration:

subchronic

Species:

rat

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEL

582 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Additional information

1.1.1        Repeated Dose Toxicity, oral

In a 14-day range-finding study with Fatty acids, sunflower-oil, conjugated, maleated, reaction products with diethanolamine, maleated tall-oil fatty acids and triethanolamine, 4 Wistar rats per sex and group received 330 and 1100mg/kg of the test substance per day via gavage(6). Clinical signs, haematological and clinical chemistry parameters were examined. Gross pathology was performed, and organ weights of adrenals, kidneys, liver and spleen determined. Body weight gain in high dose females was significantly lower. The same tendency was observed for low dose females and for absolute body weight in females of both test groups, but the differences were not statistically significant. Paralleling the effects on body weight, food consumption was not-significantly reduced in females, but not in males. Red (-6% / -8%) and white (-16% / -20%) blood cells were reduced in high dose males and females, as well as haemoglobin (-11% / -9%), haematocrit (-11% / -11%), and the number of reticulocytes (-37% / -72%). Not all differences reached statistical significance. Absolute and relative kidney weights were increased by 17-19% in high dose animals of both sexes. Relative liver weights were 36% higher in males and 26% higher in females receiving 1100mg/kg. The NOAEL based on the limited parameters examined was 330mg/kg.

 

The source substance diethanolamine was fed to 5 Fischer 344 rats per sex and group via the drinking water for 14 days at doses of 630, 1250, 2500, 5000, and 10000ppm(7). In addition to clinical signs, haematology, clinical chemistry, urinalysis, and histopathology, sperm morphology and vaginal cytology were evaluated. All females in the two highest doses and 2 high dose males died. Surviving animals of the higher dose groups exhibited depressed weight gain. Exposure to diethanolamine produced a moderate, poorly regenerative, normochromic, microcytic anemia in male and female rats of all groups, as indicated by dose-dependent decreases in erythrocyte and reticulocyte counts, MCV, haemoglobin concentration, and haematocrit. In male and female rats, serum concentrations of creatinine, total protein, UN, albumin, and bile acids (male rats) were increased by treatment with diethanolamine. The kidney weight was increased in females at 630ppm and in males at 1250ppm. At higher doses, increased incidences of renal tubular epithelial necrosis, urine concentrations of urea nitrogen, glucose, protein, and lactate dehydrogenase activity were observed. Based on the effects of the haematopoietic system, no NOAEL could be defined. The LOAEL in this study was 630ppm, corresponding to about 80mg/kg.

 

In the following 90-day main study, 10 Fischer 344 rats per sex and group were exposed to 160 (only females) 320, 630, 1250, 2500, and 5000 (only males) ppm via the drinking water(7). The study protocol was similar to OECD guideline 408. Deaths occurred in 2/10 male rats in the top dose group. Surviving animals in the higher concentration groups exhibited depressed weight gains. As already observed in the range finding study, the animals developed a moderate, poorly regenerative, microcytic, normochromic anemia in a dose-dependent manner from the lowest dose level onwards, though effects were very limited at 160 ppm. Hematologic effects were dose-dependent and included decreases in erythrocyte and reticulocyte counts, hemoglobin concentration, hematocrit, MCV, and MCH. MCV was reduced in rats at all dose levels. Hematologic effects were not associated with microscopic changes in the femoral bone marrow. No significant gross lesions attributable to diethanolamine were found at necropsy. Dose-related increases in relative kidney weights were observed in males and females. Kidney weight changes were accompanied by increases in the incidence and/or severity of nephropathy, and renal tubular cell necrosis. Nephropathy was already observed in the lowest dose in females, while changes in males aside from the weight increase didn't occur until 2500ppm. Increased nephropathy was considered a regenerative change and was supported by the observation of tubular necrosis at the higher doses. At high doses, minimal to mild demyelination of the brain and spinal cord was observed in all male and female rats in the 2500 and 5000 ppm dose groups. There were no neurologic clinical signs that could be clearly attributed to these lesions. Dose-related increases in relative liver weights occurred in male and female rats. Although the changes in liver weights were not associated with microscopic lesions in the liver, there were mild to moderate increases in serum concentrations of total bile acids in female rats in all dose groups, and in male rats in all dose groups except the lowest (320 ppm). In conclusion, the most sensitive parameter were haematological changes, and a NOAEL was not achieved. The LOAEL was set to the lowest dose for males and females, i.e., 320 ppm and 160 ppm, respectively. This corresponds to 25mg/kg in males and 14mg/kg in females.

In an extended one-generation reproductive toxicity study, rats were exposed to 100, 300, and 1000ppm of DEA in the drinking water, which corresponds to daily doses of app. 13, 38, and 128mg/kg b.w. Target organs were liver, kidney, and the hemopoietic system. The NOAEL for these effects was 100ppm (13mg/kg), which matches the results of the 90-day study described above. At the highest dose (1000ppm), demyelination and minimal to marked degeneration of nerve fibers occurred in the F1 generation, but the only behavioural difference was an impaired auditory startle response. Demye­li­na­tion was also already observed in the 90-day study starting at 2500ppm. Further details are provided

 

In a sub-chronic oral toxicity study for TEA, which was performed similarly to OECD guideline 408, 20 Cox CD rats per sex and dose were exposed to 0, 250, 500 or 1000 mg/kg bw/day in the diet for 91 days(8). For histopathology and organ weight determination, only 5 rats per sex and group were examined. No significant differences between groups were observed in haematology and organ weights. There were non-significant tendencies for increased relative liver and kidney weights starting at 500mg/kg and 1000mg/kg, respectively. Gross pathologic and histopathologic examination did not reveal any treatment-related effects. In the absence of histopathologic findings and the non-significant differences, the NOAEL was established to be 1000 mg/kg bw/day, the highest dose tested.

1.1.2        Repeated Dose Toxicity, inhalation

Since they are not relevant for the target substance for reasons stated above, local effects have been omitted from the study summaries.

 

In a study performed according to OECD guideline 413, 13 Wistar rats per sex and group were exposed to 15, 150, 400mg/m³ of DEA aerosol for 90 days on 5 days per week. No deaths or clinical signs were observed. Final body weight in high dose males was reduced by 13%. No treatment related differences occurred during FOB examinations. At 400 mg/m³, red blood cells, haemoglobin, haematocrit and mean corpuscular volume were significantly decreased in male and female rats, matching the symptoms of a mild microcytic, normochromic anaemia. Morphological examination revealed only a marginal increase in anisocytosis in the respective males.Mild increases of liver weights and serum alkaline phosphatase serum levels without histopathological findings in the mid and high concentration indicate a concentration dependent adaptive response.Histopathology revealed a moderate to severe (grade 3-4) diffuse testicular atrophy accompanied by oligozoospermia in the epididymides in 3 high dose males. Slight atrophy of the prostate was observed in 4 high dose males. Focal atrophy up to grade 3 was seen in all groups including control animals. Absolute and relative kidney weights were increased in mid and high dose animals. This was accompanied by blood in the urine of high dose animals and an increased excretion of renal tubular epithelium cells including casts in mid and high dose males. Four females each of the mid and high dose showed minimal to slight tubular hyperplasia. Because of the slight alterations of the kidneys still present in the mid dose, the NOAEL was conservatively set to 15mg/m³.

 

With the source substance TEA, repeated inhalation toxicity was investigated in a sub-acute 28 -day study performed according to OECD TG 412, in which Wistar rats (10/sex/dose) were exposed head/nose only to 0, 0.02, 0.1 or 0.5 mg/L for 6 hours/day and 5 days/weekaerosol(9). No mortality was observed. No statistically significant differences between groups were observed in body weight, haematology and clinical chemistry. Differences in grip strength were judged not substance-related because of a lack of concentration- or time-related effect. No other abnormalities were observed during neurofunctional testing. A significant difference in red blood cells was observed in males of the mid-dose group compared to controls, but since this deviation was marginal, not observed in females, and not dose-related, this finding was considered of no toxicological significance. The systemic NOAEC was established to be 0.5 mg/L, the highest dose tested.

 

1.1.3        Repeated Dose Toxicity, dermal

Since they are not relevant for the target substance for reasons stated above, local effects have been omitted from the study summaries.

 

The toxicity of DEA after repeated dermal exposure was examined in a 90-day study performed similarly to OECDguideline 411. Ten malesand female rats per group were exposed to 32, 63, 125, 250, and 500mg/kg of DEA dissolved in ethanol on 5 days per week. One high dose male and two high dose females died or were killed moribund. Final body weights were reduced in males receiving at least 250mg/kg and in females receiving 125mg/kg or more. Moderate microcytic, normochromic anemia developed in male and female rats. Decreases in red blood cells, haemoglobin, haematocrit, MCV, MCH, and reticulocytes were observed even at the lowest dose, though at this level not all changes were significant. Effects worsened dose-dependently. There were no histological changes in femoral bone marrow. Serum biochemical changes in male rats included a mild increase in ALT activity in the 3 highest dose groups. In female rats, UN, albumin, and total protein increased in all dose groups (except at the lowest dose for total protein), and total bile acids increased in the 2 highest dose groups. A mild increase in activity of ALT occurred in female rats in the highest dose group. Effects were noted for the kidneys in form of an increase in absolute and relative kidney weights in male and female rats. These weight changes were associated in females with renal tubular cell necrosis (starting at 250mg/kg), and an increased incidence of tubular mineralization (starting at the lowest dose). In males, there was no histopathological correlate. In the absence of histopathological alterations, the dose-dependent increase in live weights was considered adaptive. Sperm morphology and vaginal cytology evaluations did not show adverse effects. Based on the adverse haematological changes and kidney effects, no NOAEL could be identified. The LOAEL was set to the lowest dose of 32mg/kg.

 

In a sub-chronic dermal toxicity study, Fischer rats were treated with 0, 125, 250, 500, 1000 or 2000 mg/kg bw/day TEA on the skin, 5 days/week for 90 days (13 weeks)(10). 20 animals/sex/dose were exposed, of which 10 were used for periodic urinalysis, hematology, and clinical chemistry determinations. The remaining 10 animals were used for collection of clinical observation data, sperm morphology and vaginal cytology evaluations, necropsy with gross examination and tissue collection, and histopathological examination. No mortality was observed. Topical application of 2000 mg/kg bw resulted in a significant decrease in body weight gain. Heamtological changes were all secondary to chronic skin inflammation. Clinical chemistry findings in high dose animals of increased serum alanine and aspartate aminotransferase activities were suggestive of liver injury. However, sorbitol dehydrogenase activity, which is generally considered to be a better gauge of liver damage, was not increased, and histopathology revealed no evidence of hepatic injury. Aspartate aminotransferase has a wider tissue distribution than sorbitol dehydrogenase, and increased serum activity could be related to minor injury at another site, such as the muscle, rather than to hepatotoxicity. Additionally, some compounds can cause increases in alanine aminotransferase activity in the liver or serum without causing hepatic injury. Kidney weights increased with increasing dose in male and female rats, starting at 1000mg/kg. Dosed males had decreased urinary protein excretion which likely reflected a change in renal function or an increase in protein reabsorption, as serum protein concentrations were not affected. Although these findings suggest the possibility of protein droplet accumulation or some other form of renal dysfunction or injury, no evidence of hyaline droplet nephropathy or other histopathologic changes that might account for the weight changes was noted. In the absence of histopathological correlates, the increased kidney weights were considered as non-adverse. The NOAEL was set to 1000mg/kg, based on reduced body weight and increased aspartate aminotransferase activities at 2000mg/kg.

Justification for classification or non-classification

Effects are caused by the content of DEA. Based on its concentration of 5.5% in the registered substance, the calculated LOAELs / LOAECs after subchronic exposure exceed exceeds the ranges given in REACH CLP table 3.9.3. Additionally, based on the rules for classification of mixtures, the target substance does not need to be classified. Consequently, no classification is proposed for repeated dose toxicity for the target substance according to EU GHS, Regulation (EC) 1272/2008.