应用LC-MS/MS检测中药材中黄曲霉毒素残留量方法研究

Study on the Method for Determination of Aflatoxin Residue in Traditional Chinese Medicine by LC-MS/MS

目的:建立LC-MS/MS对中药材中黄曲霉毒素残留量的检测方法。方法:中药材粗粉甲醇提取,经免疫亲和柱净化吸附,甲醇洗脱,以甲醇/乙腈-1.0mmol乙酸铵为流动相,黄曲霉毒素含量用液相色谱-串联三重四极杆质谱测定,用电喷雾正离子模式ESI,多反应(MRM)监测。结果:黄曲霉毒素G_2、B_2在0.03～3.2ng·ml^(-1)范围内线性关系良好、黄曲霉毒素B_1、G_1在0.1～10ng·ml^(-1)范围内线性关系良好。检测限0.1ng·ml^(-1)定量限0.2ng·ml^(-1)。黄曲霉毒素回收率:86.6%～93.2%,RSD为7.1%～11.8%。结论:本法专属性强,灵敏、简便,准确,通过色谱(保留时间)及质谱特征碎片离子定性,能有效排除假阳性干扰。可作为中药中黄曲霉毒素残留量测定的方法。

To establishe the method for determination of aflatoxin residue in traditional Chinese medicine by LC-MS/ MS. Method ： The powder of traditional Chinese medicine was extraeted by methanol. The extraet phase was purified by immuno-affini- ty column and eluted by methanol solution. Determination of aflatoxin residue was analyzed by liquid chromatography （LC） and TSQ Quantum Access MAX. The positive ion mode and multiple reaction monitoring （ MRM ） were used. Methanol and aeetonitrile -1.0 mmol ammonium acetate were used as the mobile phase. Result： It showed a good linearity relationship when the content of aflatoxin G2 ,B2 was within the rang from 0. 03 ng·ml^-1 to 3.2 ng·ml^-1 and the content of aflatoxin Gl ,B1 was within the rang from O. 1 ng·m1^-1 to 10 ng·ml^-1. The detection limit and quantitative limit was 0. 1 and 0. 2 ng·ml^-1, respectively. The recovery of aflatoxin was 86. 6% - 93.2% with RSD of 7.1% -11.8%. Conclusion： The method is special, sensitive and accurate. The interference of false positive is eliminated by of eharaeteristie fragment ions qualifieation （MS） and retention time （LC）. This method may be used as the determination of aflatoxiu residue in traditional Chinese medicine.