坚果中黄曲霉毒素的光化学柱后衍生-高效液相色谱法测定

Determination of Aflatoxins in nuts by High Performance Liquid Chromatography Coupled with Post-Column Photochemical Derivatization

中文关键词:  黄曲霉毒素  坚果  光化学柱后衍生  高效液相色谱

英文关键词:aflatoxins  nuts  post-column photochemical derivatization  high performance liquid chromatography (HPLC)

中文摘要:

      目的 建立坚果中黄曲霉毒素B1、B2、G1、G2的高效液相色谱荧光检测器测定方法。方法 样品以甲醇-水(70:30, v/v)溶液匀质提取, 过黄曲霉毒素总量免疫层析亲和柱净化, 经LaChrom C18色谱柱分离和光化学柱后衍生反应器衍生后, 用带有荧光检测器的高效液相色谱仪测定。采用峰面积外标法定量坚果中黄曲霉毒素B1、B2、G1、G2含量。结果 四种黄曲霉毒素在各自的浓度范围内线性关系良好, 相关系数均大于0.999, B1、B2、G1、G2的检出限依次为0.10、0.05、0.10、0.05 μg/kg。在3个添加水平下回收率为77.5%~109.8%, 相对标准偏差为1.43%~2.71%。结论 该方法的灵敏度、准确度、精密度均符合黄曲霉毒素的检测技术要求, 适用于坚果中黄曲霉毒素B1、B2、G1、G2的日常检测。。

英文摘要:

      Objective To establish a high performance liquid chromatography (HPLC) coupled with post-column photochemical derivatization method for the determination of the aflatoxin B1, B2, G1, and G2 in nuts. Methods The sample was tested by HPLC with fluorescence detector after being extracted from metha-nol-water (70:30, v/v), purified by immunoaffinity chromatography column of aggregated aflatoxin, sepa-rated by LaChrom C18 chromatographic column, and derived from post column photochemical derivative reactor. The contents of aflatoxin B1, B2, G1, and G2 in nuts were measured using peak area external standard method. Results The calibration curves showed a good linearity in each concentration range with correlation coefficient greater than 0.999. The limits of detection (LODs) were 0.10, 0.05, 0.10, and 0.05 μg/kg for the aflatoxins B1, B2, G1, and G2 in nuts, respectively. The recovery rate was within 77.5%~109.8% at three adding levels with the relative standard deviation (RSDs) of 1.43%~2.71%. Conclusion This method is sensitive, accurate and precise which is in accordance with technical standards of the aflatoxins detection. It is proper for routine detection of the aflatoxins B1, B2, G1, and G2 in nuts.