| Enzyme-linked immuno sorbent assay(ELISA)kits for Aflatoxin B1(AFB1),Zearalenone(ZEN)and Deoxynivalenol(DON)from four manufacturers which are widely used both domestic and overseas were chosen in this project.The test was strictly carried out according to the introduction of each kind of ELISA kits.Test results from the standard sample of sensitivity,accuracy and precision,reproducibility,and result consistency(compare to results acquired from high performance liquid chromatography(HPLC))and some other indexes were compared and evaluated in this project.Expt.1.Comparisons of 4 kinds of ELISA kits for AFB1Four kinds of ELISA kits were used to determinate the concentration of AFB1 in both reference standard and real sample.Evaluating the sensitivity,accuracy,precision and reproducibility of these kits through detennining the inhibitory concentration 50(IC50),average relative error,average standard deviation,slope and intercept coefficient of standard curve.By measuring the recover of sample and the result consistency compared to HPLC analysis to assess the practicability of these kits.These observations showed that IC50 of kits A1,A2,A3 and A4 were 2.18 ?g/kg.9.42 ?g/kg,8.05 ?g/kg and 6.58 ?g/kg for reference standard,respectiely;five gradients(10 ?g/kg,25 ?g/kg,50 ?g/kg,100 ?g/kg and 200 ?g/kg)of AFB1 standards were added to this test,the average relative errors of these kits were 28.74%,63.76%,20.01%and 28.74%,respectively;the average relative standard deviations were 8.82%,38.46%,13.59%and 8.82%,respectively;the RSD of standard curve of 4 kits were 14.17%,13.01%,7.91%and 7.74%,respectively;the RSD of interception were 23.74%,54.55%,12.43%and 12.31%,respectively.For real samples,add 3 gradients(10 ?g/kg,50 ?g/kg and 100 ?g/kg)of AFB1 standards to mixed batch,the average recovery were 99.86%,39.13%,91.10%and 85.39%,respectively.In the determination of real samples,The ratio of each kit and HPLC measurement value(P)were 9.28,26.89,5.08 and 1.83,and the ratio of each kit and HPLC measurement value(S)were72.22,160.42,15.77 and 3.43 respectively.In summary,the kit from A1 has good sensitivity and recovery rate.The accuracy,precision and repeatability of A3 and A4 are better than the others.Additionally,the result consistency compared to the HPLC analysis of A4 kit is better.Expt.2.Comparisons of 4 kinds of ELISA kits for ZENFour kinds of ELISA kits were used to determinate the concentration of ZEN in both reference standard and real sample.Evaluating the sensitivity,accuracy,precision and reproducibility of these kits through determining the inhibitory concentration 50(IC50),average relative error,average standard deviation,slope and intercept coefficient of standard curve.By measuring the recover of sample and the result consistency compared to HPLC analysis to assess the practicability of these kits.These observations showed that IC50 of kits Zl,Z2,Z3 and Z4 were 270.24 ?g/kg,deletion,168.51 ?g/kg and 39.27 ?g/kg for reference standard,respectively;five gradients(50 ?g/kg.250 ?g/kg,500 ?g/kg,1000?g/kg and 2000 ?g/kg)of ZEN standards were added to this test,the average relative errors of these kits were 26.43%,146.19%,8.53%and 19.35%,respectively;the average relative standard deviations were 2.68%,8.05%,3.58%and 7.42%,respectively;the RSD of standard curve of 4 kits were 34.42%,9.13%,20.27%and 12.33%,respectively;the RSD of interception were 41.40%,167.10%,22.18%and 10.78%,respectively.For real samples,add 3 gradients(250 ?g/kg,500 ?g/kg and 1000 ?g/kg)of ZEN standards to mixed batch,the average recovery were 95.83%,295.57%,218.74%and 132.77%,respectively.In the determination of real samples,The ratio of each kit and HPLC measurement value(P)were 3.66,5.76,12.90 and 4.90,and the ratio of each kit and HPLC measurement value(S)were 12.76,21.32,18.90 and 7.61 respectively.In summary,the kit from Z4 has good sensitivity and reproducibility.The accuracy and precision of Z3 is better than the others,the recovery rate of Z1 kit is better.But,the result consistency compared to the HPLC analysis of four kits are bad.Expt.3.Comparisons of 4 kinds of ELISA kits for DONFour kinds of ELISA kits were used to determinate the concentration of DON in both reference standard and real sample.Evaluating the sensitivity,accuracy,precision and reproducibility of these kits through determining the inhibitory concentration 50(IC50),average relative error,average standard deviation,slope and intercept coefficient of standard curve.By measuring the recover of sample and the result consistency compared to HPLC analysis to assess the practicability of these kits.These observations showed that IC50 of kits D1,D2,D3 and D4 were 354.31 ?g/kg,89.79 ?g/kg,589.83 ?g/kg and 403.44?g/kg for reference standard,respectively;five gradients(200 ?g/kg,500 ?g/kg,1000?g/kg,2000 pg/kg and 4000 ?g/kg)of DON standards were added to this test,the average relative errors of these kits were 44.56%,41.16%,20.89%and 50.15%,respectively;the average relative standard deviations were 6.22%,4.65%,9.99%and 3.01%,respectively;the RSD of standard curve of 4 kits were 17.45%,8.13%,11.79%and 4.53%,respectively;the RSD of interception were 14.71%,19.64%,18.95%and 7.77%,respectively.For real samples,add 3 gradients(200 ?g/kg,1000 ?g/kg and 2000 ?g/kg)of DON standards to mixed batch,the average recovery were 63.96%,99.54%,169.17%and 152.76%,respectively.In the determination of real samples,The ratio of each kit and HPLC measurement value(P)were 6.68,3.13,14.24 and 6.68,and the ratio of each kit and HPLC measurement value(S)were 9.92,7.35,30.24 and 127.16 respectively.In summary,the sensitivity,recovery rate and result consistency compared to the HPLC analysis of D2 is better than the others,the accuracy of D3 kit is better.Additionally,the kit from D4 has good precision and reproducibility. |
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