Ion-pair high-performance liquid chromatographic separation of two thyroxine glucuronides formed by rat liver microsomes.
A simple reversed-phase ion-pair high-performance liquid chromatographic separation method has been developed for thyroxine (T4) and its glucuronide metabolites formed by liver microsomes of untreated and 3-methylcholanthrene-treated rats. Besides the phenol-T4-glucuronide, another, probably acyl-T4-glucuronide, formation has been detected. The effect of pH and temperature on the stability of the acyl-T4-glucuronide was also investigated. The lowering of pH to 2 and cooling the samples to 5 degrees C is necessary to prevent the hydrolysis of acylglucuronide, while both pH and temperature do not affect the stability of the phenol-T4-glucuronide. The retention times of T4 and phenol-T4-glucuronide are highly influenced by the pH of the mobile phase, but not that of acyl-T4-glucuronide.
Jemnitz
K
K
Department of Biochemical-Pharmacology, Central Research Institute for Chemistry, Hungarian Academy of Sciences, Budapest, Hungary.
Vereczkey
L
L
D000215:Acylation; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005965:Glucuronates / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008862:Microsomes, Liver / Q000378:metabolism*; D010636:Phenols / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature; D013974:Thyroxine / Q000302:isolation & purification* / Q000378:metabolism
D005965:Glucuronates; D010636:Phenols; D008748:Methylcholanthrene; D013974:Thyroxine