Development of an Analytical Method for VInca Alkaloids Using Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS)

Abstract

The purpose of this study was to develop and validate an analytical method for vinblastine (VBL) and deacetylvinblastine (DVBL) using liquid chromatography tandem mass spectrometry (LC/MS/MS). The goal of the study was to develop a method that is sensitive enough to quantitate less than 1.0 ng/mL in plasma. The first step in method development was to optimize the LC/MS/MS by tuning the ion path and optimizing source parameters and chromatography. After development of the LC/MS/MS method, a solid phase extraction sample preparation method was developed and optimized to increase sensitivity. Method validation was carried out through determination of the sensitivity, selectivity, precision, and accuracy of the method. This method was then applied to canine plasma samples. The LC/MS/MS method was successfully developed for detection of vinblastine and desacetylvinblastine with vinorelbine as the internal standard. The LC method utilized isocratic flow rate with atmospheric chemical ionization (APCI) and three ions per analyte in multiple reaction monitoring (MRM) mode. Calibrators in methanol resulted in a LLOQ for vinblastine of approximately 0.77 ng/mL and of deacetylvinblastine at approximately 3.6 ng/mL. The LLOQ for plasma samples after SPE was 0.4450.047 for VBL and 0.2090.01 ng/mL for DVBL. The developed method for plasma extracted with SPE and analyzed via LC/MS/MS was validated and successfully applied to canine plasma samples.