免疫亲和柱净化-高效液相色谱法测定红曲制品中桔霉素的含量

Determination of citrinin in monascus products by high performance liquid chromatography combined with immunoaffinity column

中文关键词:  桔霉素  红曲制品  免疫亲和柱  高效液相色谱法

英文关键词:citrinin  monascus products  immunoaffinity column  high performance liquid chromatography

中文摘要:

      目的 建立免疫亲和柱净化-高效液相色谱法测定红曲制品中桔霉素含量的分析方法。方法 试样经甲醇-水提取, 稀释过滤后经含桔霉素特异性抗体的免疫亲和柱净化, 洗脱液经Merk Hibar RP-18e色谱柱分离, 以乙腈:0.1%磷酸水溶液作为流动相, 采用荧光检测器测定(激发波长350 nm, 发射波长500 nm), 以外标法定量。结果 桔霉素在1~50 ng/mL范围内有良好的线性关系, 相关系数为0.99991, 方法检测限为50 μg/kg, 在50、100和200 μg/kg 3个水平的加标回收率为67.55%~89.12%, 相对标准偏差为5.43%~11.25%。结论 该方法操作简便、准确, 可适用于红曲制品中桔霉素的测定。

英文摘要:

      Objective To establish a method for the determination of citrinin in monascus products by high performance liquid chromatography combined with immunoaffinity column. Methods Samples were extracted with methanol-water solution, purified by a immunoaffinity column containing specific antibody of citrinin after dilution and filteration. The eluate was separated by Merk Hibar RP-18e column with acetonitrile and 0.1% H3PO4-water as mobile phase, and detected by fluorescence detector with the excitation wavelength of 350 nm and the emission wavelength of 500 nm. Also the external standard method was used for quantitative analysis. Results Citrinin had a good linear relationship in the range of 1~50 ng/mL with correlation coefficient of 0.99991. The limit of detection was 50 μg/kg. The recoveries of citrinin spiked at 3 levels of 50, 100 and 200 μg/kg were 67.55%~89.12%, and the relative standard deviations were 5.43%~11.25%. Conclusion This method is simple, accurate and is suitable for the determination of citrinin in monascus products.