挥发性铵盐盐析辅助液液萃取结合超高效液相色谱-串联质谱法测定酱油中4种黄曲霉毒素

Determination of 4 kinds of aflatoxins in soy sauce by volatile salting-out assisted liquid-liquid extraction combined with ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  酱油  黄曲霉毒素  盐析辅助液液萃取

英文关键词:soy sauce  aflatoxin  salting-out assisted liquid-liquid extraction

中文摘要:

      目的 建立基于挥发性铵盐盐析辅助液液萃取结合超高效液相色谱-串联质谱法检测酱油中黄曲霉毒素B1、B2、G1、G2 4种黄曲霉毒素含量的分析方法。方法 酱油样品用乙腈提取, 乙酸铵作为盐析助剂分层, 提取液加水定容后, 采用C18反相色谱柱以5 mmol/L乙酸铵溶液-甲醇/乙腈为流动相进行梯度洗脱, 采用电喷雾正离子模式检测, 外标法定量。实验考察了萃取溶剂、盐析助剂及pH对萃取效率的影响。结果 最优实验条件下, 在0.5~20.0 ng/mL范围内4种黄曲霉毒素线性良好, 相关系数均大于等于0.999, 检出限为0.01~0.04 μg/kg, 定量限为0.03~0.12 μg/kg; 对空白酱油样品进行0.8、4.0、8.0 μg/kg 3个水平的加标实验, 回收率为73.2%~94.5%, 相对标准偏差为0.9%~5.9%。结论 该方法操作简捷、成本低、准确度高, 可满足对酱油中4种黄曲霉毒素的快速准确分析要求。

英文摘要:

      Objective To establish an analytical method for the determination of aflatoxins B1, B2, G1 and G2 in soy sauce based on volatile ammonium salt salting-out assisted liquid-liquid extraction combined with ultra performance liquid chromatography-tandem mass spectrometry. Methods The soy sauce sample was extracted with acetonitrile, and ammonium acetate was used as the salting-out aid for stratification, after the extraction solution was fixed in volume by adding water, gradient elution was performed using a C18 reverse-phase chromatographic column with 5 mmol/L ammonium acetate solution–methanol/acetonitrile as the mobile phase, and electrospray positive ion mode detection was adopted, followed by external standard quantification. The effects of extraction solvent, salting-out aid and pH on extraction efficiency were investigated. Results Under the optimal experimental conditions, the linearity of the 4 kinds of aflatoxins were good in the range of 0.5?20.0 ng/mL, the correlation coefficients were all greater than or equal to 0.999, and the limits of detection were 0.01?0.04 μg/kg, and the limits of quantitation were 0.03?0.12 μg/kg; the standard addition experiments at the 3 levels of 0.8, 4.0 and 8.0 μg/kg were performed on the blank soy sauce sample, and the recovery rates were 73.2%?94.5%, and the relative standard deviations were 0.9%?5.9%. Conclusion This method is simple, low in cost and high in accuracy, and can meet the requirements of rapid and accurate analysis of 4 kinds of aflatoxins in soy sauce.