HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF MORPHINE AND ITS 3-GLUCURONIDE AND 6-GLUCURONIDE METABOLITES - IMPROVEMENTS TO THE METHOD AND APPLICATION TO STABILITY STUDIES

Improvements to previously reported methods for the determination of morphine, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in human plasma are described. The improved methods involve the use of a solid-phase extraction cartridge and a chromatographic system which uses paired-ion reversed-phase high-performance liquid chromatography with a radially compressed column. Only one cartridge is used to prepare each sample for chromatography and each cartridge may be used for at least fourteen 1-ml plasma samples. The recovery is greater than 85%. The improvements to the method of sample pretreatment and in the chromatographic conditions have allowed determination of morphine, M3G and M6G in human plasma down to 13.3 nmol/l(coefficient of variation = 9.3%), 108 nmol/l(6.6%) and 41 nmol/l(6.7%), respectively, using ultraviolet detection alone. It was shown that all three compounds were stable in plasma for up to 101 weeks when stored at -20-degrees-C.