高效液相荧光色谱法同时测定六神曲中黄曲霉毒素、玉米赤霉烯酮和赭曲霉毒素A

Simultaneous determination of aflatoxin,zearalenone,and ochratoxin A in Massa Medicata Fermentata by high-performance liquid chromatography and fluorescence detection

目的建立一种用免疫亲和柱净化-柱后光化学衍生-高效液相色谱同时检测六神曲中黄曲霉毒素、玉米赤霉烯酮和赭曲霉毒素A的方法。方法样品采用60%乙腈超声提取,免疫亲和柱净化,采用XBridge^(®)Phenyl苯基色谱柱(4.6 mm×250 mm,5μm),以乙腈和0.1%磷酸溶液为流动相,梯度洗脱,光化学衍生仪衍生,通过切换荧光波长检测。结果黄曲霉毒素、玉米赤霉烯酮和赭曲霉毒素A标准曲线的线性范围分别为0.006~0.108、0.500~2.500和0.202~1.009 ng,6种毒素的线性关系在0.9994以上,检出限分别为1.2、100.0和40.3 ng/ml,平均加标回收率为73.2%~92.3%,相对偏差为2.1%~5.4%。结论该方法具有专属性强、操作方便等特点,能够有效用于六神曲中3种毒素的同时检测和安全质量控制。

Objective To establish a method for the simultaneous detection of aflatoxin,zearalenone,and ochratoxin A in Massa Medicata Fermentata using immunoaffinity column purification-post column photochemical derivatization-high-performance liquid chromatography.Methods The sample was extracted with 60%acetonitrile ultrasound,purified with an immunoaffinity column,and treated for gradient elution with XBridge^(®)Phenyl column(4.6 mm×250 mm,5μm),and with acetonitrile and 0.1%phosphoric acid solution as the mobile phase,derivatization with a photochemical derivation instrument,and detection by switching fluorescence wavelength.Results The linear ranges of the standard curves for aflatoxin,zearalenone,and ochratoxin A were 0.006-0.108,0.500-2.500,and 0.202-1.009 ng,respectively.The linear relationships of the six toxins were above 0.9994,with detection limits of 1.2,100.0,and 40.3 ng/ml,and an average spiked recovery rate of 73.2%-92.3%.The relative deviation was 2.1%-5.4%.Conclusion This method has the characteristics of strong specificity and easy operation,and can be effectively used for the simultaneous detection and safety quality control of three toxins in Massa Medicata Fermentata.