复合免疫亲和柱净化-超高效液相色谱-串联质谱法测定衢枳壳中5种真菌毒素的含量

Determination of 5 Mycotoxins in Quzhou Aurantii Fructus by Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry with Composite Immunoaffinity Column Purification

提出了题示方法测定衢枳壳样品中黄曲霉毒素B1、B2、G1、G2和赭曲霉毒素A的含量。样品经过体积比79∶20∶1的乙腈-水-乙酸混合溶液超声提取,离心,上清液用p H 7.0磷酸盐缓冲液稀释,用复合免疫亲和柱净化,过滤,滤液中的目标物在XDB C18色谱柱上用不同体积比的体积比1∶9的甲醇-5 mmol·L^(-1)乙酸铵混合溶液和体积比9∶1的甲醇-5 mmol·L^(-1)乙酸铵混合溶液的混合溶液分离,电喷雾离子源正离子(ESI+)和多反应监测(MRM)模式检测,同位素内标法定量。结果显示:5种真菌毒素与其同位素内标质量浓度比和对应的峰面积比在一定范围内呈线性关系,检出限为0.05~0.20μg·kg^(-1)。3个加标浓度水平下阴性样品中5种真菌毒素的回收率为82.7%~97.3%,测定值的相对标准偏差(n=6)为5.2%~12%。

The method mentioned by the title was proposed to determine aflatoxin B1,B2,G1,G2,and ochratoxin A in Quzhou Aurantii Fructus samples.After ultrasonic extraction on the sample in the mixed solution composed of acetonitrile,water and acetic acid at volume ratio of 79∶20∶1,the extract was centrifuged.The supernatant was diluted by phosphate buffer solution(pH 7.0),purified with a composite immunoaffinity column,and filtered.The targets in the filtrate was separated on the XDB C18 chromatographic column using mixed solutions composed of methanol-5 mmol·L^(−1) ammonium acetate mixed solution(volume ratio of 1∶9)and methanol-5 mmol·L^(−1) ammonium acetate mixed solution(volume ratio of 9∶1)at different volume ratios,detected by the ESI+and MRM modes,and quantified by the isotope internal standard method.It was shown that linear relationships between values of the mass concentration ratio and corresponding peak area ratio of the 5 mycotoxins to their isotope internal standards were kept in definite ranges,with detection limits in the range of 0.05-0.20μg·kg^(−1).Recoveries of the 5 mycotoxins in negative samples at three spiked concentration levels ranged from 82.7%to 97.3%,and RSDs(n=6)of the determined values were in the range of 5.2%-12%.