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    免疫亲和柱净化-光化学柱后衍生-高效液相色谱法测定柏子仁中的黄曲霉毒素

    Determination of Aflatoxins in Platycladi Seeds by HPLC Combined with Immunoaffinity Column Cleanup and Post-Column Photochemical Derivatization

    • 摘要: 采用免疫亲和柱净化-光化学柱后衍生-高效液相色谱法测定中药材柏子仁中的黄曲霉毒素G2、G1、B2和B1。样品经甲醇(7+3)溶液提取,提取液经免疫亲和柱净化,用甲醇洗脱,洗脱液经黄曲霉毒素专用C18色谱柱分离,以甲醇(45+55)溶液为流动相进行洗脱,柱后光化学衍生波长为254 nm,荧光检测器的激发波长为365 nm,发射波长为440 nm。黄曲霉毒素G2、B2的线性范围均为0.125~5.0 μg·L-1,黄曲霉毒素G1、B1的线性范围均为0.50~20 μg·L-1,检出限(3S/N)在0.012~0.047 μg·L-1之间。加标回收率81.4%~105%之间,测定值的相对标准偏差(n=6)在1.6%~6.9%之间。

       

      Abstract: A method of HPLC combined with immunoaffinity column cleanup and post-column photochemical derivatization was applied to determine aflatoxins G2, G1, B2 and B1 in platycladi seeds, which was a kind of Chinese herbal medicine. The sample was extracted with methanol (7+3) solution, then the extraction was purified by immunoaffinity column and the analyte was eluted by methanol. The eluent was separated on a special C18 chromatographic column for aflatoxins with a methanol (45+55) solution as mobile phase. The wavelength of post-column photochemical derivatization was 254 nm, and the excitation wavelength and emission wavelength of fluorenscence detector were 365 nm and 440 nm, respectively. The linearity ranges of aflatoxins G2 and B2 were both 0.125-5.0 μg·L-1, and the linearity ranges of aflatoxins G1 and B1 were both 0.50-20 μg·L-1, with detection limits (3S/N) in the range of 0.012-0.047 μg·L-1. Recovery rates obtained by standard addition method were in the range of 81.4%-105% and RSDs (n=6) were in the range of 1.6%-6.9%.

       

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