Determination of Aflatoxins Content in Traditional Chinese Medicine Using Immunoaffinity Column Cleanup and by High Performance Liquid Chromatography / 环境与健康杂志

ABSTRACT

Objective To develop a method for the determination of aflatoxins B1,B2,G1 and G2 in Chinese herbal medicine.Methods The herbal medicine samples were cleaned up on immunoaffinity columns and analyzed by high performance liquid chromatography(HPLC) with fluorescence detection.For chromatographic separation,a Zorbax SB C18(4.6 mm?150 mm,5 ?m) column was employed.The separation was carried out as the mobile phase of methanol-0.01 mol/L KH2PO(44+6)with a flow rate of 0.5 ml/min at column temperature of 35 ℃.The reaction tube temperature of postcolumn derivatization system was 70 ℃.The detection was observed by fluorescence with excitation at 360 nm and emission wavelength at 425 nm.Results The calibration curve showed good linearity in the range of 12.00-300.00 ?g/L for aflatoxin B1,and 24.00-600.00 ?g/L for aflatoxin B2,G1 and G2.The lowest limits of detection for aflatoxins in Chinese herbal medicine were 0.025 ?g/L for AFB1,0.085 ?g/L for AFB2,0.060 ?g/L for AFG1,and 0.055 ?g/L for AFG2,and the correlation coefficients were ≥0.996.Recoveries were 81.7%-101.2% for aflatoxin B1,B2,G1 and G2 spiked to Fructus amomi,Radix angelicae sinensis,Rhizoma polygonati,Rhizoma atractylodis macrocephalae and Rhizoma dioscoreae at the level of 5 ?g/L,and the relative standard deviations were 0.7%-4.9% in all instances.Conclusion The method has good stability,high sensitivity and high selectivity,and it is applicable to the determination of aflatoxins in Chinese herbal medicine.