Identification and determination of phase II nabumetone metabolites by high-performance liquid chromatography with photodiode array and mass spectrometric detection.
Chromatographic analyses play an important role in the identification and determination of phase I and phase II drug metabolites. While the chemical standards of phase I metabolites are usually available from commercial sources or by various synthetic, degradation or isolation methods, the phase II drug metabolites have usually more complicated structures, their standards are in general inaccessible and their identification and determination require a comprehensive analytical approach involving the use of xenobiochemical methods and the employment of hyphenated analytical techniques. In this work, various high-performance liquid chromatography (HPLC) methods were employed in the evaluation of xenobiochemical experiments leading to the identification and determination of phase II nabumetone metabolites. Optimal conditions for the quantitative enzymatic deconjugation of phase II metabolites were found for the samples of minipig bile, small intestine contents and urine. Comparative HPLC analyses of the samples of above-mentioned biomatrices and of the same biomatrices after their enzymatic treatment using beta-glucuronidase and arylsulfatase afforded the qualitative and quantitative information about phase II nabumetone metabolites. Hereby, three principal phase II nabumetone metabolites (ether glucuronides) were discovered in minipig's body fluids and their structures were confirmed using liquid chromatography (LC)-electrospray ionization mass spectrometric (MS) analyses.
Nobilis
M
M
Institute of Experimental Biopharmaceutics, Joint Research Center of PRO.MED.CS Praha a.s. and Academy of Sciences of the Czech Republic, Heyrovského 1207, CZ-500 03 Hradec Králové, Czech Republic. nobilis@uebf.cas.cz
Holcapek
M
M
Kolárová
L
L
Kopecký
J
J
Kunes
M
M
Svoboda
Z
Z
Kvetina
J
J
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000493:pharmacokinetics* / Q000652:urine; D001646:Bile / Q000737:chemistry; D001711:Biotransformation; D002074:Butanones / Q000493:pharmacokinetics* / Q000652:urine; D002851:Chromatography, High Pressure Liquid; D003626:Data Display; D005966:Glucuronidase / Q000737:chemistry; D007422:Intestines / Q000737:chemistry; D013058:Mass Spectrometry; D000077430:Nabumetone; D021241:Spectrometry, Mass, Electrospray Ionization; D013429:Sulfatases / Q000737:chemistry; D013552:Swine; D013556:Swine, Miniature
D000894:Anti-Inflammatory Agents, Non-Steroidal; D002074:Butanones; D013429:Sulfatases; D005966:Glucuronidase; D000077430:Nabumetone