Determination of aflatoxins in nelumbinis semen by immunoaffinity column clean-up and HPLC-FLD with on-line post-column photochemical derivatization and LC-MS/MS confirmation / 中国中药杂志

ABSTRACT

<p><b>UNLABELLED</b>To determine the contents of aflatoxin B1, B2, G1 and G2 in Nelumbinis Semen using on-line post-column photochemical derivatization-HPLC-FLD method and verify the method by LC-MS method.</p><p><b>METHOD</b>The samples were extracted with MeOH-H2O (80 20) and purified with inmunoaffinity column, aflatoxins were analyzed by HPLC-FLD with post-column photochemical derivatizaton. The positive samples were further confirmed by LC-MS/MS.</p><p><b>RESULT</b>On optimum conditions, aflatoxin B1, G1 ranging 0.3-30 mg x L(-1) showed a good linear relationship with aflatoxin B2, G2 ranging 0.09-9.0 mg x L(-1) with r >0.999 9. The recoveries ranged between 86.7% and 99.1%, with RSDs all bellow 4.87%. LOD of aflatoxin B1, B2, G1 and G2 were 0.08, 0.03, 0.10, 0.03 microg x kg(-1), respectively. Among 20 Nelumbinis Semen samples, 14 were found to contain aflatoxin B1 ranging from 0.40 to 586 microg x kg(-1). The total content of aflatoxin B1, B2, G1 and G2 were between 0.40 and 602.5 microg x kg(-1). By LC-MS/MS method, the same fragment ions were founded in samples and the control group at the same retention times, ruling out the possibility of false positive samples.</p><p><b>CONCLUSION</b>The method is simple, highly sensitive and reproducible for the determination of aflatoxins in Nelumbinis Semen.</p>