Simultaneous Quantification of Four Indole Alkaloids in Catharanthus roseus Cell Line C20hi by UPLC-MS

An ultra-performance liquid chromatography/mass spectrometry method to simultaneously quantify vindoline, catharanthine, serpentine and ajmalicine in Catharanthus roseus cell line C20hi is reported. Samples were extracted with 1% acetic acid, basified to pH 10 with ammonia, then extracted with ethyl acetate, dried, reconstituted with methanol–1% acetic acid water solution (1:1, v/v) and analyzed using an acetonitrile–0.1% formic acid gradient as the mobile phase. Detection was carried out by electrospray ionization mass spectrometry in the positive-ion mode with selective ion monitoring. The analysis of one sample was achieved in 6 min. The limits of detection were 0.46 – 0.70 ng/ml in cell samples, and 0.10 – 0.16 ng/ml in medium samples. The linearity of detection was over the wide range of 1.00 – 6250.0 ng/ml. Intra- and inter-day accuracies (recovery 88.0 – 111.8%) and precision (RSD 1.25 – 7.81%) showed the performance of the assay. This method provides a more sensitive and high-throughput technique to quantify the four alkaloids in large amount of samples, and will be helpful in high-production cultivar screening.