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Studies of the biliary excretion and metabolites of the antioxidant ethoxyquin, 6-ethoxy-2,2,4-trimethyl-1,2-dihydroquinoline in the rat.

1. Biliary excretion and metabolites of ethoxyquin, and gastro-intestinal absorption of ethoxyquin were studied in rat. 2. An average of 28 and 36% of the dose of 14C following intragastric administration of [14C]ethoxyquin was recovered in the bile of bile-duct cannulated rats in 12 and 24 h, respectively. 3. By g.l.c.-mass spectrometry, 75 to 85% of the 14C excreted in the 12 h bile was identified as unchanged ethoxyquin, and the following metabolites were isolated and identified: 8-hydroxy-ethoxyquin, hydroxylated 8-hydroxy-ethoxyquin, 6-ethoxy-2,2,4-trimethyl-8-quinolone, hydroxylated 6-ethoxy-2,2,4-trimethyl-8-quinolone, 6-ethoxy-2,4-dimethylquinoline and 2,2,4-trimethyl-6-quinolone. 4. Three groups of rats were used in the biliary excretion experiments, and the effect of standardization of experimental conditions was demonstrated. Infusion of sodium taurocholate following bile-duct cannulation did not affect the biliary excretion kinetics of ethoxyquin. 5. Only about 3% of the radioactivity administered was absorbed from the gastrointestinal tract via the lymphatic pathway in thoracic-duct connulated rats within 24 h. It was concluded that ethoxyquin was absorbed primarily by the portal route.

Adsorption

Influence of ethoxyquin on the utilization of selenium by the chick.

Experiments showed that commonly used concentrations of dietary ethoxyquin (6-ethoxy-2,2,4-trimethyl-1,2-dihydroxyquinoline) spare the selenium requirement of the vitamin E-deficient chick according to the following function: log Y = -0.0011X - 0.7741, where Y = Se requirement (ppm) and X = dietary ethoxyquin (ppm). The basis of the sparing effect appeared to be metabolic; (a) ethoxyquin was effective in alleviating exudative diathesis when fed (physiologically and chronologically) separately from selenium, and (b) ethoxyquin was effective in promoting increases in the plasma of the selenium-containing enzyme, glutathione peroxidase.

Animals

The distribution of 14C-ethoxyquin in rat.

Adult male rats were given the antioxidant 14C-ethoxyquin by oral intubation and were sacrificed at various time intervals from 0.5 hr to 6 days following administration of the drug. The distribution pattern was studied by whole-body autoradiography and liquid scintillation counting. The isotopelabelled antioxidant was distributed throoughout most tissues and the blood at 0.5 hr after administration. The highest radioactivity throughout the experimental period was observed in the liver, the kidney, the gastrointestinal tract and the adipose tissue. No activity was observed in the brain and the central nervous system. Of the dose ingested 2.2 and 0.2% were found in the liver at 0.5 hr and 6 days respectively following dosing. The hepatic peak in radioactivity was measured at 8 hrs and after 6 days 7.5% of this level was still present in the liver. Six days after administration residues of ethoxyquin and metabolites were also present in the kidney cortex, the intestines, the lung, various adipose tissue and blood.

Animals

Nondestructive distinction between aflatoxin B1 and ethoxyquin in thin-layer chromatography.

A rapid and simple method has been developed for the nondestructive distinction between aflatoxin B1 and the feed antioxidant, ethoxyquin. These two chemicals exhibit similar RF values in certain solvent systems and produce a similar bluish fluorescence under long UV (366 nm) radiation. The method involves the in situ generation of fluorescence spectra of the respective thin-layer chromatography spots. Since it is nondestructive, the method affords ancillary study of the separated aflatoxins.

Aflatoxins

Teratologic assessment of maleic hydrazide and daminozide, and formulations of ethoxyquin, thiabendazole and naled in rats.

Teratogenicity studies were conducted in rats treated orally from days 6-15 of gestation with single daily doses of 400-1600 mg/kg of maleic hydrazide, 300-1000 mg/kg daminozide, 125-500 mg/kg ethoxyquin or thiabendazole, or 25-100 mg/kg naled. Dams were killed on the 22nd day of gestation, and fetuses were evaluated by routine teratologic methods. No adverse effect was related to any treatment other than an increased incidence of anomalous fetuses at the highest dose (500 mg/kg) of thiabendazole.

Animals

Studies on the metabolism of the antioxidant ethoxyquin, 6-ethoxy-2,2,4-trimethyl-1,2-dihydroquinoline in the rat.

1. The metabolism of ethoxyquin (6-ethoxy-2,2,4-trimethyl-1,2-dihydroquinoline) in rat has been investigated. Urinary metabolites were identified by combined g.l.c.-mass spectrometry. 2. The major metabolic reaction was de-ethylation which gave rise to 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline and an oxidation product, 2,2,4-trimethyl-6-quinolone. Other reactions were hydroxylation to four different hydroxylated metabolites and one dihydroxylated metabolite. A total of 95% of the dose (100 mg/kg) was accounted for.

Animals

Ethoxyquin (EMQ) residues in Atlantic salmon measured by fluorimetry and gas chromatography (GLC).

Salmons of two different sizes (group I; 22--30 g, group II; 600--1450 g) were tested for ethoxyquin (EMQ) residues after fed ad libitum a diet supplied with the therapeutic level of 900 ppm EMQ for 2 months. The sums of residues of EMQ and possible metabolites having similar fluorescence characteristics were determined by a fluorimetric method. The percentage of unmetabolized EMQ present was estimated using a gas chromatographic-mass spectrometric (GLC-MS) method measuring only unchanged EMQ. For both methods the detection limit was 0.1 ppm. EMQ residues were also fluorimetrically estimated in muscle tissue of small salmons fed a diet preserved with 150 ppm EMQ for 2 months. At termination of feeding the diet supplied with 900 ppm EMQ, residues measured fluorimetrically in muscle tissue of salmons in both groups were averaging 0.7 ppm. Approximately one third was unchanged EMQ. From salmons of group II blood, kidney and hepatic tissue were also analyzed at termination of feeding and residues averaging 0.3, 0.8 and 1.8 ppm respectively were estimated. The elimination of EMQ from muscle tissue is illustrated in Fig. 1. No residues could be detected 9 days following termination of feeding the therapeutic level of EMQ. In muscle tissue of salmons fed the diet preserved with 150 ppm EMQ, residues ranging from 0.1 to 0.3 ppm of EMQ and possible metabolites were found in 9 of 10 samples obtained from 20 salmons in group I. For an evaluation of EMQ used as a feed additive (150 ppm), further studies were recommended. The diets used in the experiment contained 88 and 830 ppm EMQ respectively measured fluorimetrically at the end of the experiment. Approximately 88% of the residues in the feed was unchanged EMQ. It is suggested that most information regarding the hygienic aspects of residues in muscle tissue is obtained by fluorimetry which is therefore recommended for determination of EMQ residues in tissues. The GLC-MS method, however, is recommended for estimation of EMQ levels in feed.

Animal Feed

Inhibition of thiabendazole metabolism in the rat.

1. A single oral dose of desmethylimipramine (80 mg/kg) administered to rats inhibited the hepatic microsomal hydroxylation of thiabendazole (45%), aniline (30%), biphenyl (30%) and ethylmorphine (15%) in vitro at 5 h after dosage; there was no decrease in cytochrome P-450 or b5. 2. A single oral dose of ethoxyquin (200 mg/kg) to rats inhibited the hepatic microsomal hydroxylation of thiabendazole (65%), aniline (40%) and biphenyl (40%) in vitro at 1 h after dosage; inhibition was less at 5 h. There were no changes in the contents of cytochromes P-450 and b5. 3. The max. plasma concn. of thiabendazole occurred 2--4 h after oral dosing (50--200 mg/kg) to rats. Thiabendazole (100 mg/kg) administered orally 30 min after oral ethoxyquin (400 mg/kg) or thiabendazole (200 mg/kg) administered orally 30 min after oral desmethylimipramine (80 mg/kg) delayed absorption of the thiabendazole and resulted in markedly markedly decreased plasma concentration of the anthelmintic. 4. Simultaneous administration of ethoxyquin (300 mg/kg) potentiated the anthelmintic effect of thiabendazole (750 mg/kg) on the helminth parasite, Nematospiroides dubius, in the mouse. Desmethylimipramine showed no similar potentiation.

Animals