Optimization of a liquid chromatography-tandem mass spectrometry method for the quantification of traces of taxanes in a Corylus avellana cell suspension medium

Abstract

Since the recent discovery of taxol and other taxanes in Corylus avellana, this plant species has attracted interest as a potential new source of these compounds. However, its low taxane content in comparison with Taxus spp. has restricted research to analytical identification or global quantitation. A feasible and sensitive method based on liquid chromatography-tandem mass spectrometry using a triple quadrupole analyzer was developed for the analysis of taxol and four other taxanes in a Corylus avellana cell suspension medium. Taxanes were extracted from the cell culture medium with dichloromethane and analyzed using electrospray ionization and quantified by multiple-reaction monitoring mode. Methanol and matrix-matching calibration curves using docetaxel as the internal standard were analyzed. Linearity was confirmed over the whole calibration range (0.3-2.1 μg mL-1). The inter- and intra-day precision of taxanes ranged from 80% to 120% and the recovery rates were higher than 80%. Limits of detection were between 0.24-38 ng mL-1 and the limits of quantification were between 0.8-125 ng mL-1. The low detection and quantitation values obtained allowed us to detect small quantities of the released taxanes (120 ng mL-1 of B, 151 ng mL-1 of CF and 105 ng mL-1 of T), which correspond to about 0.5 ng mL-1 of each taxane, in the 20 mL Corylus avellana cell suspension culture medium extracted, even at the beginning of the culture. These results were confirmed by high resolution mass spectrometry. This journal is