Identification of sulfate and glucuronic acid conjugates of the 5-hydroxy derivative as major metabolites of 2-amino-3-methylimidazo[4,5-f]quinoline in rats.

New metabolites of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), a potent mutagen and carcinogen formed during cooking of meat or fish, have been identified and quantitated in the urine and bile of rats. Administration was either by a pulse gavage dose of 40 mg/kg [2-14C]IQ or by dietary intake of 300 ppm IQ for 6 weeks. The metabolites were isolated by high-performance liquid chromatography and quantitated by radioactivity. They were then characterized by their resistance or sensitivity to hydrolytic enzymes or acid hydrolysis, by nuclear magnetic resonance and mass spectrometry, or coinjection with a synthetic sample. A minor metabolite was the IQ N-glucuronide. A major metabolite was formed by hydroxylation of IQ at the 5-position; it was present in urine and bile and was conjugated as the glucuronide or sulfate ester, which together accounted for about 40% of urinary or biliary metabolites. The unconjugated compound partially adsorbs onto the high-performance liquid chromatographic columns used. The amounts of 5-OH-IQ present as conjugates in urine or bile were similar, irrespective of mode of administration. Thus, hydroxylation of IQ on carbon 5 followed by type II conjugation reactions yields quantitatively important metabolic products.

Metabolism of 2-amino-3-methylimidazo[4,5-f]quinoline in the male rat.

The metabolism of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) was studied in the male rat using the radiochemical labels 14C and 3H at positions 2 and 5 of the molecule, respectively. Adult male Fischer 344 rats were administered [2-14C]IQ or [5-3H]IQ by oral gavage at dose levels of 20 or 40 mg/kg body weight. Rats were also given [2-14C]IQ in the diet at a dose level of 300 ppm for 2 days and after administration of unlabelled IQ (300 ppm) in the diet for approximately 6.5 wk for an additional 2 days. In the initial 48 hr following oral administration of 20 or 40 mg [2-14C]IQ/kg body weight, about 40-50% radioactivity was recovered in the urine, and about 30-38% radioactivity was recovered in the faeces. In the initial 72 hr following consumption of [2-14C]IQ (300 ppm) in the diet about 26% radioactivity was recovered in the urine and about 61% radioactivity was recovered in the faeces. Following cannulation of the bile ducts, rats administered a single dose of [2-14C]IQ (40 mg/kg body weight) by oral gavage excreted about 15% of the administered dose in the bile over a period of 2 days. Urine from rats given [2-14C]IQ contained three main polar metabolites that included a glucuronide, a sulphate ester and IQ sulphamate, and a number of less polar metabolites that included IQ, 2-acetylamino-3-methylimidazo[4,5-f]quinoline, 2-aminoimidazo[4,5-f]quinoline and 2-amino-3,6-dihydro-3-methyl-7H-imidazo[4,5-f]quinoline-7-one (7-OH-IQ). Administration of [2-14C]IQ by oral gavage or in the diet gave the same metabolites, but in different amounts. In the faeces of rats given [2-14C] by oral gavage, IQ-sulphamate was the major metabolite in the polar fraction. Non-polar metabolites similar to those found in the urine were also present, but in different amounts. A major, non-polar faecal metabolite, 7-OH-IQ was probably formed as a result of the activity of the intestinal bacterial flora. In rats given a single gavage dose of [2-14C]IQ, excretion of metabolites was higher in the urine and lower in the faeces compared with that in animals fed [2-14C]IQ in the diet. One polar metabolite present in the urine, IQ-sulphamate (39%), was found at considerably higher levels in rats dosed orally with IQ compared with those fed IQ (less than 6%). Thus, IQ is extensively metabolized to give a number of polar and non-polar metabolites, the amounts of which depend, in part, on the mode of dosing.

Comparison of XAD-2 column and blue cotton batch techniques for isolation of metabolites of 2-amino-3-methylimidazo[4,5-f]quinoline.

In studies on the metabolism of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in the rat, two methods were used to concentrate IQ and its metabolites in urine: XAD-2 columns and blue-cotton extraction. These methods were compared as to the total recovery of 14C-label and the results of high-performance liquid chromatography (HPLC) of the main metabolites. In the HPLC analysis, three major peaks in the polar region and two adjacent peaks in the nonpolar region having radioactivity were found in the urine from rats given 14C-IQ. XAD-2 columns adsorbed approximately 65% and blue cotton 23% of the applied isotope from the urinary metabolites. Both methods efficiently adsorbed and totally released the relatively nonpolar compounds in a mixture of metabolites, including the compound administered, IQ. However, they failed to retain the more polar metabolites. Thus, both the XAD-2 column and blue cotton adsorption techniques may be useful mainly to concentrate nonpolar compounds from larger volumes of aqueous solutions.