同位素标记-高效液相色谱-串联质谱法测定中药材中8种真菌毒素

目的建立同位素标记-高效液相色谱-串联质谱法(HPLC-MS/MS)测定中药材中黄曲霉毒素B1、B2、G1、G2、伏马毒素B1、赭曲霉毒素A、杂色曲霉毒素、玉米赤霉烯酮8种真菌毒素残留的分析方法。方法样品经甲醇-甲酸-水溶液(79∶1∶20,V/V)提取后,通过适当稀释和过滤,采用Thermo Hypersil GOLD色谱柱(100 mm×2.1 mm,1.9μm)分离,0.1%甲酸乙腈-0.1%甲酸水为流动相,电喷雾正负离子扫描模式,同位素内标法定量。结果在该实验条件下,8种真菌毒素在浓度0.33μg/L^500μg/L时,线性关系良好,r均>0.999;检出限(LOD)为0.1μg/kg^5.0μg/kg;3种不同浓度的添加水平下,8种真菌毒素的平均回收率为80.3%~108.8%,相对标准偏差(RSD)为0.1%~9.6%。结论将方法用于24种中药材中真菌毒素残留的检测,测定结果表明该方法具有前处理方式简单,检验方法准确、可靠、灵敏度高、检测速度快的优点,可广泛应用于中药材中8种真菌毒素的残留检测。

Determination of 8 mycotoxins in Chinese medical materials by isotope labeling-high performance liquid chromatography-tandem mass spectrometry

Objective An isotope labeling-high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) method was developed for the determination of eight mycotoxin residues in traditional Chinese medicines:aflatoxin B1,B2,G1,G2,fumonisin B1,ochratoxin A,miscellaneous aspergillus toxin and zearalenone.Methods The samples were extracted by methanol-formic acid-water solution(79∶1∶20,V/V),diluted and filtered properly.The separation of 8 mycotoxins was carried out with a Thermo Hypersil GOLD chromatographic column,using 0.1% formic acid acetonitrile-0.1% formic acid solution.8 mycotoxins were determined in positive and negative electro-spray ionization mode.Quantitative analysis was conducted by isotope internal standard method.Results Under the optimized conditions,the calibration curve showed good linearity within 0.33 μg/L-500 μg/L,with correlation coefficients greater than 0.999.The limits of detection(LODs) were within 0.1 μg/kg-5.0 μg/kg.The recoveries of 8 mycotoxins at three levels were within 80.3%-108.8%,and RSDs were within 0.1%-9.6%.Conclusion The method has been applied to the determination of mycotoxin residues in24 kinds of Chinese medicinal materials.The results show that the methods were simple,accurate,reliable,sensitive,rapid,and can be widely used for the residual detection of 8 mycotoxins residues in Chinese medicinal materials.