免疫亲合柱净化高效液相色谱检测啤酒中赭曲霉毒素A

建立了检测啤酒中赭曲霉毒素A(OTA)免疫亲合柱净化高效液相色谱法。该方法为将脱气后的啤酒样品用15%NaCl和2%NaHCO_3的水溶液稀释,然后通过Ochra Test免疫亲和柱净化,采用C18(5μm,250×4.60 mm)色谱柱,流动相为乙腈-水-乙酸(体积比为99:99:2),流速0.900 mL/min;激发波长333 nm;发射波长477 nm;柱温35℃;进样量100μL;外标法定量。方法所采用的标准曲线有良好的线性关系,检出限为0.1 ng/mL,加标回收率为95%～106%,对不同浓度的样品相对标准偏差RSD为2.43%～8.32%。该方法操作简便、准确,回收率高,精密度良好,重现性好,可用于啤酒中赭曲霉毒素A的测定,具有实际应用价值。

Determination of Ochratoxin A in Beer by Immunoaffinity Column Clean-up and High Performance Liquid Chromatography

Immunoaffinity Column clean-up and high performance liquid chromatography method(HPLC) for the determination of OTA in beer have been established.20mL of degassed beer wete diluted with 4mL of a water solution containing 15 %NaCl and 2 % NaHCO_3,then were cleaned up through OchraTest column. Separation was carried out on a column of C_(18) (5μm,250×4.60 mm)at 35℃with acetonitrile-water-acetic acid(99:99;2,v/v) as mobile phase,elution flow rate 0.90mL/min.The detection of excitation wave- length was 333nm and the emission wavelength was 470nm,quantified by external standard method.The ex- perimental results showed a good linear relationship between the peak area and the concentration,the recov- eries for OTA were 95%～106%,and the relative standard deviation was 2.43%～8.32% for different kinds of beer.The results show that the method is simple,rapid,accurate and applicable for the determination of OTA in beer.