Simultaneous Quantification of Four Indole Alkaloids in Catharanthus roseus Cell Line C20hi by UPLC-MS
Simultaneous Quantification of Four Indole Alkaloids in Catharanthus roseus Cell Line C20hi by UPLC-MS
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DOI：
10.2116/analsci.27.433
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发表时间：
2011-04-01
影响因子：
1.6
通讯作者：
Hu, Zhibi
中科院分区：
文献类型：
--
作者：
He, Lihong;Yang, Li;Hu, Zhibi
建立了超高效液相色谱/质谱法同时测定长春花C20 hi细胞中文多灵、长春质碱、蛇纹石和阿吉马林的方法。样品用1%乙酸提取，用氨水碱化至pH 10，然后用乙酸乙酯提取，干燥，用甲醇-1%乙酸水溶液（1：1，v/v）复溶，并使用乙腈-0.1%甲酸梯度作为移动的相进行分析。通过电喷雾电离质谱法以正离子模式进行检测，并进行选择性离子监测。一个样品的分析在6分钟内完成。检测限为0.46 - 0.70 ng/ml的细胞样品和0.10 - 0.16 ng/ml的培养基样品。在1.00 ~ 6250.0ng/ml范围内线性关系良好。日内和日间准确度（回收率88.0 - 111.8%）和精密度（RSD 1.25 - 7.81%）显示了该方法的性能。该方法为大批量样品中4种生物碱的定量分析提供了一种更灵敏、更高通量的方法，为高产品种的筛选提供了依据。
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.