免疫亲和柱净化-高效液相色谱柱后光化学衍生- 荧光检测器检测食品中黄曲霉毒素

Determination of aflatoxins in food by immunoaffinity chromatography-high performance liquid chromatography fluorescence detector coupled with post-column photochemical derivatization

中文关键词:  黄曲霉毒素  食品  高效液相色谱法  柱后光化学衍生

英文关键词:aflatoxins  food  high performance liquid chromatography  post-column photochemical derivatization

中文摘要:

      目的 建立一种免疫亲和固相萃取柱净化-高效液相色谱柱后光化学衍生-荧光检测器同时测定食品中黄曲霉毒素B1、B2、G1和G2的方法。方法 以甲醇-水(70:30, V:V)为提取溶剂, 采用高速均质提取, 并经过黄曲霉毒素免疫亲和柱净化。结果 经月旭公司Welch Ultimate? XB-C18色谱柱(250 mm×4.6 mm, 5 μm)分离后使用光化学衍生器进行柱后衍生, 并采用带荧光检测器的高效液相色谱仪检测, 流动相为甲醇/水(45 : 55, V:V)。黄曲霉毒素B1、B2、G1和G2线性范围在0.3~50.0 μg/L之间, 线性相关系数均大于0.999, B1、B2、G1和G2检出限分别为0.15 μg/kg、0.05 μg/kg、0.15 μg/kg、0.05 μg/kg。在3 个加标浓度下大米、花生和瓜子等试样的回收率在80.7%~92.6% 之间; 相对标准偏差(RSD)在2.04~3.87%之间。结论 该方法的灵敏度、准确度和精密度均符合黄曲霉毒素的检测技术要求, 且简便快速, 适用于食品中黄曲霉毒素B1, B2, G1和G2的准确测定。

英文摘要:

      Objective To establish an immunoaffinity chromatography-high performance liquid chromatography (HPLC) fluorescence detector coupled with post-column photochemical derivatization method for de-termination of the aflatoxin B1, B2, G1, and G2 in food. Methods The sample was tested by HPLC with fluorescence detector after being homogenizer extracted from methanol-water (70:30, V:V), purified by immunoaffinity chromatography column of aggregated aflatoxin, separated by Welch Ultimate?XB-C18 HPLC column, eluted with a mobile phase consisting of methanol and waters (45:55, V:V) and derived from post column photochemical derivative reactor. The content of aflatoxin B1, B2, G1, and G2 in food was 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 within linear range 0.3~50.0 μg/L. The limits of detection (LODs) were 0.15, 0.05, 0.15 and 0.05 μg/kg for the aflatoxins B1, B2, G1, and G2 in food, respectively. The recovery rate was within 80.7 %~92.6 % at 3 adding levels with the relative standard deviation (RSDs) of 2.04~3.87%. 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 food.