Simultaneous determination of free mycophenolic acid and its glucuronide in serum of patients under mycophenolate mophetil therapy by ion-pair reversed-phase liquid chromatography with diode array UV detection

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Antonella ArestaGiuseppe Grandaliano

Abstract

An high performance liquid chromatography (HPLC)-UV method for the simultaneous determination of the free forms of mycophenolic acid (MPA) and its phenol glucuronide (MPAG) in human serum samples was developed for the first time. Chromatographic separation was performed on octadecylsilane based stationary phase in combination with a mobile phase of methanol/buffered tetrabutylammonium (TBA) salt mixture. Sample pretreatment consisted of an ultrafiltration step followed by clean-up/enrichment on a C(18) solid-phase extraction (SPE) cartridge. Average recoveries of (99.7 +/- 0.2)% and (64.1 +/- 6.9)% for free MPA and MPAG, respectively, were estimated in the concentration range from 0.5 to 10 microg/ml. The within-day and between-days coefficients of variation were 0.4 and 0.8% for free MPA (0.1 microg/ml spiking level) and 0.8 and 1.6% for free MPAG (5 microg/ml spiking level), respectively. The linear ranges for free MPA and MPAG were 0.06-1 and 0.2-10microg/ml, respectively. Detection limits of 4 and 17 ng/ml for free MPA and MPAG were estimated in spiked serum. The same HPLC method was also capable of a simultaneous determination of the total concentration of MPA and MPAG when coupled to a proper sample pretreatment step. The...Continue Reading

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Citations

Oct 13, 2015·Drug Metabolism and Personalized Therapy·Bing ChenBei-ming Xu
May 8, 2007·Clinical Biochemistry·Fawzy A Elbarbry, Ahmed S Shoker
Mar 14, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xavier CussonneauRoselyne Boulieu
Aug 26, 2014·World Journal of Gastroenterology : WJG·Hao Chen, Bing Chen

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