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药材中黄曲霉毒素的免疫亲和层析净化高效液相色谱法
Determination of Aflatoxins Content in Traditional Chinese Medicine Using Immunoaffinity Column Cleanup and by High Performance Liquid Chromatography

作  者: ; ; ; (梁美琼);

机构地区: 广东食品药品职业学院

出  处: 《环境与健康杂志》 2010年第1期66-69,共4页

摘  要: 目的建立高效液相色谱法测定中药材砂仁中黄曲霉毒素B1、B2、G1、G2的方法。方法样品经免疫亲和层析净化,色谱柱为Zorbax SB C18柱(4.6mm×150mm,5μm),流动相为甲醇-0.01mol/L的KH2PO(44+6),流量为0.5ml/min,柱温为35℃,激发波长为360nm,发射波长为425nm,柱后衍生反应器温度为70℃,进行HPLC检测。结果黄曲霉毒素B1在12.00~300.00μg/L范围内、黄曲霉毒素B2、G1、G2在24.00~600.00μg/L范围内时,回归方程呈良好的线性关系,检出限分别为0.025、0.085、0.060、0.055μg/L,r≥0.996。该方法的平均回收率为81.7%~101.2%,RSD为0.7%~4.9%。结论该方法灵敏度高,选择性好,方法稳定,可用于中药材中黄曲霉毒素的检验。 Objective To develop a method for the determination of aflatoxins B1, B2, G2 and G: in Chinese herbal medicine. Methods The herbal medicine samples were cleaned up on immnnoaffinity columns and analyzed by high performance liquid chromatography (HPLC) with fluorescence detection. For chromatographic separation, a Zorbax SB Cis (4.6 mm×150 mm,5 μm) column was employed. The separation was carried out as the mobile phase of methanol-0.01 mol/L KH2PO4( 4+6 ) with a flow rate of 0.5 ml/min at column temperature of 35 ℃. The reaction tube temperature of posteolumn 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 μg/L for aflatoxin Bt, and 24.00-600.00 μg/L for aflatoxin B2,Gt and G2. The lowest limits of detection for aflatoxins in Chinese herbal medicine were 0.025 μg/L for AFBI, 0.085 μg/L for AFB2,0.060 μg/L for AFGl,and 0.055 μg/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, Rhizorrti, atractylodis macrocephalae and Rhizorna dioscoreae at the level of 5μg/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.

关 键 词: 色谱法 高效液相 黄曲霉素 中药材 免疫亲和层析

领  域: [理学] [理学]

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