牛乳中黄曲霉毒素B_1和M_1检测方法的比较

Comparison of methods for detecting aflatoxins B_1 and M_1 in milk

比较了免疫亲和柱-高效液相色谱法(HPLC)和酶联免疫吸附法(ELISA)测定牛乳中黄曲霉毒素B1(AFB1)和M1(AFM1)的回收率、方法检出限及精密度。结果表明,HPLC法测定AFB1、AFM1的检测限分别为0.02和0.01μg·L-1,ELISA法测定AFB1、AFM1的最低检出限分别为0.05和0.02μg·L-1;阴性牛乳试样AFB1(0.003、0.006、0.012、0.024μg·kg^(-1))和AFM1(0.005、0.01、0.025、0.05μg·kg^(-1))的回收率试验表明,HPLC法测定AFB1与AFM1的回收率分别为77.58%~82.81%和77.51%~82.23%,变异系数分别为1.93%和3.51%;ELISA法测定AFB1与AFM1回收率分别为77.28%~79.11%和75.40%~76.34%,变异系数分别为2.20%和3.80%,两种方法测定AFB1的回收率差异不显著(P>0.05),当AFM1添加浓度大于0.025μg·kg^(-1)时,HPLC法回收率显著高于ELISA法(P<0.05);综上,两种方法灵敏度高、重复性好,HPLC法测定高浓度AFM1时准确性优于ELISA法。

Immunoaffinity column-high performance liquid chromatography（HPLC）and enzyme-linked immunosorbent assay（ELISA）methods were adopted to determine aflatoxins（AF）B1 and M1 in milk.The recovery,detection limit,and precision of these two methods were compared in this study.The results showed that,for HPLC,the detection limits of AFB1 and AFM1 were 0.02 and 0.01μg·L-1,respectively,while for ELISA,the detection limits of AFB1 and AFM1 were 0.05 and 0.02μg·L-1,respectively.Standard solutions of AFB1（0.003,0.006,0.012,and 0.024μg·kg-1）and AFM1（0.005,0.01,0.025,and 0.05μg·kg-1）were added into blank milk samples and the results of recovery experiments indicated that,for HPLC,the recoveries of AFB1 and AFM1 were 77.58%82.81% and 77.51%82.23% with relative standard deviations of 1.93%and 3.51%,respectively.For ELISA,the recoveries of AFB1 and AFM1 were 77.28%79.11% and 75.40%76.34% with relative standard deviations of 2.20%and 3.80%,respectively.The recovery of AFB1 did not significantly differ between the two methods（P〉0.05）.However,the recovery of AFM1,when the amount added was more than 0.025μg·kg-1,was significantly higher by HPLC than by ELISA（P〈0.05）.In conclusion,both the tested methods were highly accurate,with a high sensitivity and good repeatability.At higher concentrations of AFM1,the accuracy of HPLC was better than that of ELISA.