Determination of a novel epothilone D analog (AV‐EPO‐106) in human plasma using ultra‐performance liquid chromatography–tandem mass spectrometry

Abstract

A novel ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method has been established for the determination of a newly synthesized epothilone D analog (AV-EPO-106) in human plasma. The plasma samples were prepared by liquid-liquid extraction with cold tert-butyl methyl ether. The chromatographic separation was achieved within 5 min on a C(18) column with water-methanol (10:90, v/v) as mobile phase at a flow-rate of 0.8 mL/min. Mass transition of m/z 568.2 to 386.1 was measured for AV-EPO-106 in positive atmospheric pressure chemical ionization mode. A detailed validation of the method was performed as per the USFDA guidelines. For AV-EPO-106 at the concentrations of 1.0, 5.0 and 10.0 microg/mL in human plasma, the absolute extraction recoveries were 86.17, 85.24 and 85.69%, respectively. The linear quantification range of the method was 0.10-20.0 microg/mL in human plasma with linear correlation coefficients greater than 0.999. The intra-day and inter-day accuracy for AV-EPO-106 at the levels of 1.0, 5.0 and 10.0 microg/mL in human plasma fell in the ranges of 98.25-100.47 and 94.19-97.25%, and the intra- and inter-day precision were in the ranges of 4.75-6.30% and 8.89-10.45%, respectively. The method was successfully applied to quantify AV-EPO-106 in human plasma to determine the half-life of this compound in human plasma.

Simultaneous determination of epothilone D and its hydrolytic metabolite in human plasma by high performance liquid chromatography–tandem mass spectrometry for pharmacokinetic studies

“…Some analytical methods were reported for epothilone D and epothilone D analog, to be used high performance liquid chromatography (HPLC) [1] and ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) [10], respectively. In this method, the low limit of quantitation is 0.2 ng/ml which is decreased 500-fold as compared to previous methods, the sample preparation is precipitated protein with acetonitrile which is more simpler and faster than that in liquid-liquid extraction [1,10]. Only 100 l of plasma is required for the sample pretreatment procedures, this is so important for clinical study, especially for clinical patients.…”

“…For its further study, it is necessary to develop an analytical method for the determination of such a compound, especially in biological fluids. Some analytical methods were reported for epothilone D and epothilone D analog, high performance liquid chromatography (HPLC) [1] and ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) [10]. And a few papers were reported preclinical pharmacology of epothilone D. In order to satisfy the urgent request of clinical study, in this research, we develop a LC/MS/MS method that employed a simple acetonitrile precipitation procedure for analyte recovery and that allowed sensitive and rapid quantitation of epothilone D and its hydrolytic metabolite in human plasma simultaneously.…”