QuEChERS前处理结合高效液相色谱法及高效液相色谱-串联质谱法检测葡萄酒中3种赭曲霉毒素

Detection of three ochratoxins in wine by QuEChERS combined with high performance liquid chromatography and high performance liquid chromatography-tandem mass spectrometry

目的建立QuEChERS净化技术结合高效液相色谱法及高效液相色谱-串联质谱法检测葡萄酒中赭曲霉毒素A(OTA)、赭曲霉毒素B(OTB)和赭曲霉毒素C(OTC)3种赭曲霉毒素的方法。方法对QuEChERS净化条件和高效液相色谱及高效液相色谱-串联质谱检测条件进行优化,葡萄酒样品先用乙腈-冰乙酸(90∶10,V/V)进行酸化稀释,离心后取上清液经C_(18)+SiO_2+MgSO_4净化剂组合净化过滤,待测样液经液相色谱C_(18)柱(4.6 mm×250 mm,5μm)梯度洗脱分离后,分别采用荧光检测器和串联质谱电喷雾技术,在多反应监测(MRM)模式下,实现同时对葡萄酒中OTA、OTB和OTC三种赭曲霉毒素的定性和定量分析。结果方法最低定量限(LOQ)均为2.0μg/kg,相对标准偏差(RSD)为2.01%～7.52%(n=6)。结论该方法操作方便、灵敏度高、重现性好,适用于日常监管工作中对葡萄酒中赭曲霉毒素的检测。

Objective Method were developed to detect three ochratoxins including ochratoxin A（OTA）, ochratoxin B（OTB） and ochratoxin C（OTC） in wine via QuEChERS combined with high performance liquid chromatography（QuEChERS-HPLC） and HPLC-tandem mass spectrometry（HPLC-MS/MS）. Methods The wine samples were diluted with acetonitrile-glacial acetic acid（90∶10, V/V） and then purified with a modified QuEChERS using a scavenging agent combination of C_（18）, SiO_2 and MgSO_4 after centrifugation. The separation was performed on C_（18） column（4.6 mm×250 mm, 5 μm） after the gradient elution on a liquid chromatography. Three ochratoxins contaminants in wine were analyzed by HPLC-FLD and HPLC-MS/MS-ESI in the positive ionization under the multiple-reaction monitoring（MRM） mode. Results By these method, the limit of quantification（LOQ） were 2.0 μg/kg and the relative standard deviation（RSD） was between 2.01%-7.52%（n=6）. Conclusion The developed methods were proved to be simple, sensitive, reproducible and were suitable for the determination of ochratoxins in wine in daily supervision.