高效液相色谱-串联质谱法同时测定配制酒中5种紫杉烷

Simultaneous Determination of 5 Kinds of Taxanes in Blending Liquor by Using High Performance Liquid Chromatography-tandem Mass Spectrometry

建立了同时测定配制酒中紫杉醇、三尖杉宁碱、巴卡亭Ⅲ、7-表-紫杉醇和10-脱乙酰基巴卡丁Ⅲ(10-DAB)5种紫杉烷的高效液相色谱-串联质谱(HPLC-MS/MS)法。样品经40℃氮吹浓缩,二氯甲烷与水(6∶3,v∶v)萃取净化,采用Thermo Hypersil Gold C18色谱柱(100 mm×2.1 mm,3μm)分离,以乙腈和0.1%甲酸水进行梯度洗脱,质谱正离子多反应监测(MRM)模式测定。结果显示,5种紫杉烷在0~120 ng/mL范围内线性关系良好,相关系数r≥0.9982;紫杉醇、7-表-紫杉醇、巴卡亭Ⅲ检出限为4μg/kg,三尖杉宁碱、10-DAB检出限为2μg/kg;回收率为71.84%~87.68%,相对标准偏差为1.33%~8.82%。该方法操作简单、灵敏度高,能对配制酒中5种紫杉烷进行定性、定量检测,从而辅助鉴别配制酒中非法添加红豆杉的行为,为开展配制酒中红豆杉非法添加的整治工作提供技术支撑。

A method for the determination of 5 taxanes including taxol,cephalomannine,baccatin III,7-epi-taxol and 10-deacetylbaccatin III(10-DAB)in blending liquor was developed with optimized high performance liquid chromatography tandem mass spectrometry conditions. The taxanes were concentrated by nitrogen sweeping with thermostatic waterbath at 40 ℃,purified by dichloromethane(water:dichloromethane,6∶3,v∶v),and separated by Thermo Hypersil Gold C18 chromatographic column(100 mm×2.1 mm,3 μm). Acetonitrile and 0.1% formic acid aqueous solution were used as mobile phase,and the positive ion mode with multiple reaction monitoring(MRM)was used to detect taxanes. The results showed that the 5 taxanes exhibited good linear relationships in the range of 0~120 ng/mL with correlation coefficients(r)of more than 0.9982;the limits of detection for taxol,7-epi-taxol and baccatin III were 4 μg/kg,and for cephalomannine and 10-deacetylbaccatin III were 2 μg/kg,respectively. The recoveries ranged from 71.84% to 87.68% with the relative standards deviations of 1.33%~8.82%. This simple and sensitive method could determine the five kinds of taxanes in blending liquor qualitatively and quantitatively,and assist in the identification of illegal addition of Taxus chinensis in blending liquor,and provide useful technical support for the regulation of illegal addition of Taxus chinensis in blending liquor.