Analytical Sciences

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

Lihong HE,*,** Li YANG,* Aizhen XIONG,* Shujuan ZHAO,* Zhengtao WANG,* and Zhibi HU*

*The Moe Key Laboratory of Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P. R. China
**Department of Cell Biology, Medical College of Soochow University, Suzhou 215123, P. R. China

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.

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