在线免疫亲和固相萃取-高效液相色谱法快速测定食品中赭曲霉毒素A

Rapid and sensitive determination of ochratoxin A in foods by on-line automatic immunoaffinity solid-phase extraction with high performance liquid chromatography

中文关键词:  在线固相萃取  食品安全  赭曲霉毒素A  免疫亲和  高效液相色谱法

英文关键词:on-line solid-phase extraction  food safety  ochratoxin A  immunoaffinity  high performance liquid chromatography

中文摘要:

      目的 建立在线免疫亲和固相萃取-高效液相色谱快速测定食品中赭曲霉毒素A(ochratoxin A, OTA)的分析方法。方法 谷物、豆类样品采用乙腈-水(80:20, V:V) 提取, 葡萄酒类样品采用NaHCO3/NaCl/水溶液(0.4:15:100, m:m:V)提取, 3% Tween-20(m:V)溶液稀释后, 直接经RIDA? CREST ICE在线免疫亲和固相萃取系统净化-高效液相色谱荧光检测OTA含量。以乙酸铵-乙腈为流动相洗脱, 流速1.0 mL/min, C18色谱柱 (150 mm×4.6mm, 5 μm) 分离测定。结果 方法检出限分别为0.020 μg/kg(豆类、谷物)和0.050 μg/kg(葡萄酒类); 在OTA的浓度为12.5~500 ng/L的范围内线性关系良好, 相关系数r2为0.9995; 日内加标回收率为87.4%~109.8%, 相对标准偏差0.50%~4.80%。结论 该方法分析速度快, 灵敏度高, 重现性好, 可满足快速准确测定大批量食品样品中OTA含量的需要。

英文摘要:

      Objective To establish a analytical method for determination of ochratoxin A in food by on-line automatic immunoaffinity solid-phase extraction with high performance liquid chromatography (HPLC). Methods The samples of grain and legumes were extracted with acetonitrile water (80:20, V:V). The wine samples were extracted with NaHCO3/NaCl/ aqueous solution (0.4:15:100, m:m:V), 3% Tween-20 (m:V) solution was diluted, and the OTA content was detected directly by the RIDA @CREST ICE on-line immuno affinity solid phase extraction system purification and high performance liquid chromatography fluorescence. The sample solution was eluted with ammonium acetate and acetonitrile as mobile phase at a flow rate of 1.0 mL/min, separated and determined by C18 column (150 mm×4.6 mm, 5 μm). Results The detection limits were 0.020 μg/kg (legumes, grains) and 0.050 μg/kg (wine), and the linear relationship was good in the range of 12.5-500 ng/L of OTA, the correlation coefficient r2 was 0.9995, the recoveries within the day were 87.4%-109.8%, and the relative standard deviations were 0.50%-4.80%. Conclusion The proposed method is rapid, simple, repeatable and sensitive, which is suitable for high throughput determination of OTA in food.