免疫亲和层析净化-高效液相色谱法测定大米中桔霉素

HPLC Determination of Citrinin in Rice with Purification by Immunoafinity Chromatography

大米样品经甲-醇10 mmol·L^-1磷酸溶液（70＋30）提取,提取液稀释过滤后经CitriTestTM免疫亲和柱层析净化后用高效液相色谱法测定其桔霉素的含量。以Kromasil 100-10 C18色谱柱（250 mm×4.6 mm,5μm）为分离柱,甲酸（1＋99）溶液与乙腈以体积比1比1混合的溶液作流动相,采用荧光检测,激发波长为350 nm,发射波长为500 nm。必要时采用液相色谱-串联质谱法对结果作验证。桔霉素的质量浓度在5.0-500.0μg·L^-1围内与其峰面积呈线性关系,测定下限（10S/N）为10.0μg.kg^-1。方法的回收率为73%-76%,相对标准偏差（n=8）为7.5%-11.8%。

Citrinin was extracted from rice sample with a mixture of methanol and 10 mmol·L^-1 phosphoric acid（70＋30）,and the extract obtained was diluted and purified by passing through the immoumoaffinity chromatographic column Citri TestTM column.In the HPLC analysis,Kromasil 100-10 C18 column（250 mm×4.6 mm,5 μm） was used as separation column,and a solution of dil.formic acid（1＋99） and acetonitrile mixed in the ratio of 1 to 1 by volume was used as mobile phase.Fluorescence detection at wavelengths of（λex） 350 nm and（λem） 500 nm was adopted in the determination.Results were confirmed if necessary by hyphenation of LC-MS/MS.Linear relationship between values of peak area and mass concentration of citrinin was obtained in the range from 5.0 to 500.0 μg·L-1,with lower determination limit（10S/N） of 10.0 μg·kg-1.Values of recovery and RSD′s（n=8） of the method were found in the range of 73%-76% and 7.5%-11.8%,respectively.