Zhang Lin, Gai Qing-Hui, Zu Yuan-Gang, Yang Lei, Ma Yu-Liang, Liu Yang
State Engineering Laboratory of Bio-Resource Eco-Utilization, Northeast Forestry University, Harbin 150040, China; Engineering Research Center of Forestry Bio-preparation, Ministry of Education, Northeast Forestry University, Harbin 150040, China; Key Laboratory of Forest Plant Ecology, Ministry of Education; Northeast Forestry University, Harbin, 150040, China.
State Engineering Laboratory of Bio-Resource Eco-Utilization, Northeast Forestry University, Harbin 150040, China; Engineering Research Center of Forestry Bio-preparation, Ministry of Education, Northeast Forestry University, Harbin 150040, China; Key Laboratory of Forest Plant Ecology, Ministry of Education; Northeast Forestry University, Harbin, 150040, China.
Chin J Nat Med. 2014 Oct;12(10):786-93. doi: 10.1016/S1875-5364(14)60120-5. Epub 2014 Oct 31.
To establish a method to simultaneously determine the main five alkaloids of Catharanthus roseus for trace samples, a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) analysis method was developed.
The five Catharanthus alkaloids, vinblastine, vincristine, vinleurosine, vindoline, and catharanthine were chromatographically separated on a C18 HPLC column. The mobile phase was methanol-15 nmol·L(-1) ammonium acetate containing 0.02% formic acid (65 : 35, V/V). The quantification of these alkaloids was based on the Multiple Reaction Monitoring (MRM) mode.
This method was validated, and the results achieved the aims of the study. The intra- and inter-day precision and accuracy of the five alkaloids were within 1.2%-11.5% (RSD%) and -10.9%-10.5% (RE%). The recovery rates of the five alkaloids of samples were from 79.9% to 91.5%. The five analytes were stable at room temperature for 2 h, at 4 °C for 12 h, and at -20 °C for two weeks. The developed method was applied successfully to determine the content of the five alkaloids in three plant parts of three batches of C. roseus with a minute amount collected from three regions of China.
The HPLC-ESI-MS/MS method can be used for the simultaneous determination of five important alkaloids in trace C. roseus samples.
为建立一种可同时测定长春花中主要5种生物碱的痕量样品分析方法，开发了一种高效液相色谱-电喷雾电离-串联质谱（HPLC-ESI-MS/MS）分析方法。
长春花中的5种生物碱，长春碱、长春新碱、长春罗新、文多灵和长春质碱，在C18高效液相色谱柱上进行色谱分离。流动相为甲醇-含0.02%甲酸的15 nmol·L⁻¹醋酸铵（65∶35，V/V）。这些生物碱的定量基于多反应监测（MRM）模式。
该方法经过验证，结果达到了研究目的。5种生物碱的日内和日间精密度及准确度分别在1.2%-11.5%（相对标准偏差）和-10.9%-10.5%（相对误差）范围内。样品中5种生物碱的回收率为79.9%至91.5%。这5种分析物在室温下2小时、4℃下12小时以及-20℃下两周内均稳定。所开发的方法成功应用于测定从中国三个地区采集的微量的三批长春花三个植物部位中5种生物碱的含量。
HPLC-ESI-MS/MS方法可用于同时测定痕量长春花样品中的5种重要生物碱。
