免疫亲和层析净化高效液相色谱测定赭曲霉毒素A的方法研究

Determination of Ochratoxin A in Foods with Immunoaffinity Column Clean-up Combined with High Performance Liquid Chromatography

试样经甲醇-水或碳酸氢钠-水溶液提取,提取液稀释过滤后经过含有赭曲霉毒A特异性抗体的免疫亲和柱层析净化,洗脱液用C18(150×4.60mm,5μm)色谱柱分离,荧光检测器测定。流动相为乙腈:水:乙酸为99:99:2;激发波长333nm;发射波长477nm;柱温35℃;进样量100μl;外标法定量,结果表明,色谱峰面积与含量之间有良好的线性关系,方法的检出限为0.2μg/kg,加标回收率为90.3%～106%,对不同浓度的样品相对标准偏差RSD为1.75%～4.32%。该方法操作简便、准确,回收率高、精密度良好、重现性好,可用于谷物、酒类、调味料等产品中赭曲霉毒素A的测定。

A rapid and accurate method for the determination of ochratoxin A （OTA） in foods was established. Samples were extracted by methanol- water or NaHCO3 solution. After dilution the extarcts, flowed through a OchraTest column containing specific antibody of OTA. The column was eluted with PBS buffer, OTA buffer, pure water, and methanol in turn. The methanol eluate was separated on C18 （150 × 4.60 mm,5 μm） at 35 ℃ with acetonitrile- water- acetic acid （99：99：2, V/V） as mobile phase at the elution flow rate 0.90 ml/min. The excitation wavelength of fluorescence detector was 333 nm and the emission wavelength was 477 nm. It was found that there is a good linear relationship between the peak area and the concentration. The recoveries of OTA are 90.3% to 106%, and the relative standard deviations arel.75% to 4.32% for different kinds of food sample. The method is simple, rapid, accurate and applicable for the determination of OTA in foods.