BITERTANOL EXPLANATION Bitertanol was first evaluated by the 1983 JMPR which allocated a temporary acceptable daily intake of 0-0.005 mg/kg bw and required, by 1987, metabolism studies to clarify the metabolic pathway of bitertanol in mammals, an oral toxicity study in the dog with minimum duration of one year and a chronic toxicity and carcinogenicity study in rats at appropriate dosage (Annex 1, WHO/FAO, 1984). Relevant data has been submitted for evaluation by the present Joint Meeting and is summarized in this monograph addendum. EVALUATION FOR ACCEPTABLE INTAKE BIOLOGICAL DATA Biochemical Aspects Absorption, Distribution and Excretion Groups of 5 male and 5 female Wistar rats received either a single oral dose of 14C-phenyl-radiolabelled bitertanol solution in ethylene glycol (100 mg/kg or 100 mg/kg), a single intravenous dose (100 mg/kg) or 14 daily oral doses (100 mg/kg) Followed by a single oral dose of 14C-radiolabelled bitertanol (100 mg/kg). Urine and fetes were collected 6 and 24 hours after dosing and subsequently at 24 hour intervals until sacrifice, 7 days later. Serial blood samples were collected during this period. Faecal excretion of radioactivity following intravenous administration indicates that biliary excretion was predominant. In each case 14C-radioactivity was mostly excreted in the faeces. Urinary excretion amounted to about 4-11% of administered dose whilst expired radioactivity was not detectable. Total recoveries of radioactivity were high, about 92-104%. Pharmacokinetic analysis indicated that absorption of an oral dose of bitertanol followed a first order pattern at the single and repeated low dose (100 mg/kg) but not at the higher dose (1000 mg/kg), suggesting saturation of absorption, distribution or elimination processes. The highest tissue levels detected were in liver (17.9 ppm, female) and kidney (5.9 ppm, male) of the high-dose groups. Tissue residues totalled about 0.2-0.4% of administered radioactivity (Puhl & Hurley, 1983). Metabolism Alternative pathways for the metabolism of absorbed bitertanol is shown in the Figure. Hydroxylation of the para-position of the biphenyl and of a methyl group of the sec-butyl moieties gave rise to phenolic (1) and diol (2) metabolites. Although one diastereomer of the latter was detectable, it underwent oxidation to the corresponding butanoic acid (4) and subsequent ring hydroxylation (5). The para-hydroxylated diol (6) was also detected as was 4,4-dihydroxybiophenyl (7). Other hydroxylated metabolites (8,9,10) were also tentatively identified (Puhl & Hurley, 1983). Toxicological Studies Special Study on Hepatotoxicity Groups of 10 male and 10 female Wistar rats received by gavage 0, 30, 100 and 300 mg/kg bitertanol (95.8% purity) suspended in distilled water with Cremorph EL daily for 14 days. On sacrifice, blood was collected for detailed hematological and clinical chemistry examination and liver samples taken for detailed histopathology and enzyme studies. During treatment female rats of the mid- (1/10) and high-dose (9/10) groups exhibited hair loss. In some cases, weight loss occurred in mid- and high-dose rats while low-dose females had reduced bodyweight gain in comparison with control animals. Female rats had a dose-related tendency to mild thrombocytosis which was significant at mid- and high-doses only. Serum gamma-glutamyl transpeptidase and bilirubin levels were slightly raised in high dose female rats. There were no gross pathological findings at necropsy. Liver weights tended to be increased at mid- and high-doses, especially in females. Histopathology showed slight to moderate bile duct proliferation with peribiliary infiltration of monocytes or polynucleocytes at mid- and high-doses. Sometimes these changes were accompanied by the presence of parenchymal Councilman bodies or, more occasionally, mitoses. The high-dose hepatocytes were occasionally swollen with finely granular cytoplasm. Fatty infiltration was not remarkable. Results of the in vitro studies were consistent with induction of hepatic microsomal enzymes: cytochrome P-450 content increased in a dose-related manner, especially in males. Aminopyrene N-demethylase activity was increased in high-dose males and females and in mid-dose males while O-demethylase activity was increased in mid-and high-dose males and high-dose female rats. Hepatic triglyceride content was not influenced by bitertanol treatment. The results of this study indicate that bitertanol caused mild hepatotoxicity with modest induction of hepatic microspinal activity in rats at 100 and 300 mg/kg (Mihail & Luckhaus, 1985). Short-term Studies Dogs Groups of 4 male and 4 female beagle dogs received bitertanol (95-97.3% purity) for two years in the diet at 0, 10, 40 or 160 ppm. Although the control dogs exhibited a greater increase in bodyweight than did treated dogs, no relationship with bitertanol treatment was observed. Good nutritional status was maintained by dogs of all groups and there were no significant differences between groups in food and water consumption. No abnormalities were seen on examination or selected reflexes, body temperature and pulse rate. However, 3 of 8 high-dose dogs developed bilateral cataracts. Urinalysis and hematological examination, conducted at approximately 3 month intervals, revealed no abnormalities. However, serum alanine amino-transferase and alkaline phosphatase were elevated at 40 and 160 ppm. At necropsy, the mean liver weight of high-dose dogs only were markedly increased. Histological examination showed mild to moderate vacuolation of adrenal zona reticularis epithelia at and above 40 ppm. Based on this finding, the no observed effect level indicated by this study is 10 ppm in the diet (Hoffman & Groening, 1983). In another study, groups of 6 male and 6 female beagle dogs were fed 0, 3 and 25 ppm bitertanol (96.3-96.7%) in the diet for 12 months. A high dose group was maintained on a diet containing 200 ppm bitertanol for 20 months to permit continued ophthalmoscopic examination. Treatment had no apparent effect on the dogs' behaviour, appearance or nutritional status. Food consumption and bodyweight gain were also unaffected. Physical examinations of pulse rates, body temperature and selected reflexes were unremarkable. Ophthalmoscopy, conducted at 3 monthly intervals in the control, low- and mid-dose groups was normal. However, in the high-dose group, one dog developed severe bilateral lenticular cataracts, observable from week 58. Four other animals exhibited slight lens opacification by week 85. In addition, dogs of this group also had signs of conjunctivitis with nasociliary discharge and incrustation. Intermittent elevation of serum hepatic alanine amino-transferase, glutamate dehydrogenase and alkaline phosphatase also occurred in the high-dose dogs. Other usual clinical chemical, hematological and urinalysis parameters were not affected by treatment in any group. At necropsy there was no grossly abnormal pathology. The adrenal weights of high-dose dogs were apparently increased in comparison to controls and lipoid vacuolations were observed in zona reticularis epithelia. The results of this study accord with the previous findings (Hoffman & Vogel, 1983). Rabbits In a repeated-dose study, bitertanol (95.8% pure) in a aqueous suspension (0.5 ml) was applied to the intact or abraded dorsal and lateral skin of groups of 6 male and 6 female New Zealand rabbits 6 hours daily 5 times per week for 3 weeks at 0, 50 and 250 mg/kg. The animals were inspected for signs of dermal irritation 24 hours after each treatment. Hematological, clinical chemical and urinalysis parameters were investigated at termination and the animals necropsied for further examination. Dermal exposure to bitertanol had no apparent effect on appearance or behaviour, bodyweight or survival of the rabbits. Transient erythema developed for a few days at the exposure areas, initially with abraded skin but later with intact skin. There was no effect on skin-fold thickness, hematology, clinical chemistry or urinalysis parameters. At necropsy, there were no gross pathological findings. Histopathological examination showed a slight epidermal thickening in exposed areas only of all treated animals. This study indicates that bitertanol is a weak skin irritant only. In vitro liver preparations showed no evidence of induction of microsomal N-demethylation or O-demethylation or Cytochrome P-450 content (Heimann & Vogel, 1984). COMMENTS The 1983 JMPR requested data to clarify the metabolic pathway of bitertanol and additional toxicological data from a one year dog study and chronic and carcinogenicity studies in rats. In rats, bitertanol is extensively metabolized, primarily by ring and side chain hydroxylation, oxidation to the corresponding carboxylic acid and also by ether cleavage. Pharmacokinetic studies demonstrate that it is absorbed more completely at lower (100 mg/kg bw) than at higher (1000 mg/kg bw) doses. Tissue retention of absorbed material is low and biliary excretion predominates. Data evaluated by the 1983 JMPR show that short-term feeding of bitertanol to rats produced body weight depression and hepatotoxicity at 600 and 2400 ppm, (equivalent top 60-240 mg/kg bw/day). Body weight depression was seen in a 90-day study at 300 ppm (30 mg/kg bw/day). On chronic administration, bitertanol retarded the growth of rats at 500 ppm (25-50 mg/kg bw/day). In view of the reduced absorption of bitertanol at high doses that has now been demonstrated, and because of the above mentioned toxicity to rats at relevant doses, the meeting considered that further chronic or carcinogenicity studies in rats would be unlikely to yield new information of toxicological significance. An additional study confirms that dogs are more susceptible to bitertanol than rats. Feeding studies of one and two years' duration show the formation of cataracts, conjunctivitis and mild hepatotoxicity, with a no-observed-adverse-effect level at 10 ppm. TOXICOLOGICAL EVALUATION LEVELS CAUSING NO TOXICOLOGICAL EFFECT Rat: 20 ppm in the diet, equivalent to 1 mg/kg bw/day. Dog: 10 ppm in the diet, equivalent to 0.25 mg/kg bw/day. ESTIMATE OF ACCEPTABLE DAILY INTAKE FOR MAN 0-0.003 mg/kg bw. STUDIES WHICH WILL PROVIDE INFORMATION VALUABLE IN THE CONTINUED EVALUATION OF THE COMPOUND Observations in man. REFERENCES Heimann, K.G. & Vogel, O., 1984. KWG 0599 Subacute Study of Dermal Toxicity to Rabbits. Unpublished study from Bayer AG Institute of Toxicology Report No. 12571. Submitted to WHO by Bayer AG, Leverkusen, FRG. Hoffman, K. & Groening, P., 1983. KWG 0599/Chronic Toxicity Study on Dogs (2-Year Feeding Experiment). Unpublished study from Bayer AG Institute of Toxicology Report No. 12307. Submitted to WHO by Bayer AG, Leverkusen, FRG. Hoffman, K. & Vogel, O., 1983. KWG 0599 (Bitertanol)/Second Chronic Toxicity Study with Dogs on Oral Administration. Unpublished study from Bayer AG Institute of Toxicology Report No. 12328. Submitted to WHO by Bayer AG, Leverkusen, FRG. Mihail, F. & Luckhaus, G., 1985. KWG 0599 (Bitertanol)/Subacute Oral Toxicity Study with Special Attention to Effect on the Liver. Unpublished study from Bayer AG Institute of Toxicology Report No. 13230. Submitted to WHO by Bayer AG, Leverkusen, FRG. Puhl, R.J. & Hurley, J.B., 1983. The Absorption, Excretion and Metabolism of Baycor TM-phenyI-UL-C14 by Rats. Unpublished study from Mobay Chemical Corporation Agri. Chem. Division. Report No. 85832. Submitted to WHO by Bayer AG, Leverkusen, FRG.
See Also: Toxicological Abbreviations Bitertanol (Pesticide residues in food: 1983 evaluations) Bitertanol (Pesticide residues in food: 1984 evaluations) Bitertanol (JMPR Evaluations 1998 Part II Toxicological)