Frommherz L, Kintz P, Kijewski H, Köhler H, Lehr M, Brinkmann B, Beike J
Institute of Legal Medicine, University Hospital Münster, Röntgenstrasse, Münster, Germany.
Int J Legal Med. 2006 Nov;120(6):346-51. doi: 10.1007/s00414-005-0071-9. Epub 2006 Jan 6.
A new specific and sensitive LC-MS-MS method for the detection of taxine B and isotaxine B, the main toxic pseudo-alkaloids from yew (Taxus sp.), in biological samples (blood, urine, gastric content) was developed. Biological samples were prepared for LC-MS-MS by means of solid-phase extraction (SPE) procedure and yielded a recovery of 86%. Chromatographic separation was achieved using an RP(18) column. Detection of taxine B and isotaxine B was performed using multiple reaction monitoring with m/z 584.2 as precursor ion, i.e. M+H, of both isomers and m/z 194.3 and m/z 107.1 as product ions after collision-induced dissociation. Docetaxel was applied as internal standard. The method was fully validated for the analysis of blood samples. Linearity was proven in the range from 0.1-500 ng/g. The limit of detection and the limit of quantitation are 0.4 and 2 ng/g, respectively. The method was applied to the determination of taxine B and isotaxine B in four fatal cases (two humans, two horses) with suspected yew intoxication. Blood levels were 105, 168, 174 and 212 ng/g.
建立了一种新的特异性和灵敏性的液相色谱-串联质谱法，用于检测生物样品（血液、尿液、胃内容物）中的紫杉碱B和异紫杉碱B，这两种物质是红豆杉属植物中的主要有毒假生物碱。生物样品通过固相萃取（SPE）程序进行处理以用于液相色谱-串联质谱分析，回收率为86%。使用反相（RP）（18）柱实现色谱分离。采用多反应监测模式检测紫杉碱B和异紫杉碱B，两种异构体均以m/z 584.2作为前体离子，即M+H，碰撞诱导解离后的产物离子分别为m/z 194.3和m/z 107.1。多西他赛用作内标。该方法已针对血样分析进行了全面验证。在0.1 - 500 ng/g范围内证明了线性关系。检测限和定量限分别为0.4和2 ng/g。该方法应用于4例疑似红豆杉中毒的死亡病例（2例人类、2例马匹）中紫杉碱B和异紫杉碱B的测定。血药浓度分别为105、168、174和212 ng/g。
