酶联免疫吸附法测定云南铁皮石斛中黄曲霉毒素B1

Determination of aflatoxin B1 in Dendrobium candidum of Yunnan province by enzyme linked immunosorbent assay method

中文关键词:  铁皮石斛  黄曲霉毒素B1  酶联免疫吸附法  高效液相色谱法

英文关键词:Dendrobium candidum  aflatoxin B1  enzyme-linked immunosorbent assay  high performance liquid chromatography

中文摘要:

      目的 建立酶联免疫吸附法测定铁皮石斛中黄曲霉毒素B1(afatoxin B1, AFB1)的分析方法, 并研究酶联免疫吸附法 (enzyme-linked immunosorbent assay, ELISA)和高效液相色谱法 (high performance liquid chromatography, HPLC)测定云南铁皮石斛中AFB1的结果差异性。方法 用甲醇+水溶液(50+50, V/V)提取石斛中AFB1, 采用酶联免疫吸附法和高效液相色谱法同时检测云南铁皮石斛(铁皮枫斗)中AFB1, 进行准确性、精密度和回收率等方法学实验, 比较2种方法的差异性。结果 ELISA法在测定范围内AFB1与其对应的吸光度值之间呈良好的线性关系, 回归方程Y=-34.798X+18.134, 相关系数r2为0.9967; 添加1.0、4.0、10.0 μg/kg 3个浓度的平均加标回收率为85.3%～94.7%, 相对标准偏差为2.57%～4.88%。2种方法的结果符合率为93.2%~117%, 相对标准偏差均小于10%。结论 ELISA法具有操作简单、检测速度快、灵敏、特异性好等优点, 可同时检测大批量样品中AFB1的含量。

英文摘要:

      Objective To establish a method for the determination of aflatoxin B1 in Dendrobium candidum of Yunnan province by enzyme-linked immunosorbent assay (ELISA) and to study the difference between ELISA and high performance liquid chromatography(HPLC) for the determination of AFB1 in Dendrobium candidum from Yunnan province. Methods Aflatoxin B1 (AFB1) was extracted from Dendrobium candidum with methanol and aqueous solution (50+50, V/V). AFB1 in Dendrobium candidum was detected by ELISA and HPLC, and the accuracy, precision and recovery rate of aflatoxin B1 in Dendrobium candidum were compared. Results There was a good linear relationship between AFB1 and its corresponding absorbance value in the detection range of ELISA. The regression equation was Y=-34.798X+18.134, and the correlation coefficient r2 was 0.9967. The average recoveries and relative standard deviations were 85.3%?94.7% and 2.57%?4.88% with the addition standard of 1.0, 4.0 and 10.0 μg/kg. The coincidence rates of ELISA and HPLC were 93.2%?117%, and the relative standard deviations were less than 10%. Conclusion ELISA has the advantages of simple operation, fast detection speed, sensitivity and good specificity. It can simultaneously detect the content of AFB1 in a large number of samples.