高效液相色谱-荧光检测法测定水产品中孔雀石绿代谢物残留量

Determination of Residual Amounts of Malachite Green and Its Metabolite in Aquatic Products by High Performance Liquid Chromatography with Fluorescence Detector

水产品中残留的孔雀石绿经硼氢化钾还原为其代谢物隐色孔雀石绿,采用高效液相色谱对隐色孔雀石绿进行了检测。经均匀化处理的试样用乙腈在pH 4.5的乙酸盐缓冲介质中提取,所得提取液用二氯甲烷液-液分配后,用酸性氧化铝柱进行分离及净化。以Agilent SB-C18色谱柱(250 mm×4.6 mm,5μm)为分离柱,乙腈和乙酸盐缓冲溶液以体积比80比20的混合溶液为流动相,用荧光检测器检测。孔雀石绿的质量浓度在1.0～500.0μg.L-1范围内与其峰面积呈线性关系,检出限(3S/N)为0.46μg.kg-1。方法的回收率在78.0%～95.2%之间,相对标准偏差(n=6)在3.01%～5.98%之间。试验结果表明,孔雀石绿转化为隐色孔雀石绿的转化率在99.1%～100.8%之间。

The residual malachite green（MG） in aquatic products was reduced to leuco-malachite green（LMG） with KBH4,and determined by high performance liquid chromatography（HPLC）.MG was extracted from the homogenized tissue sample with acetonitrile in an acetate buffer medium of pH 4.5,separated and purified by liquid-liquid partition with CH2Cl2 and acidic alumina column.The Agilent SB-C18（250 mm×4.6 mm,5 μm） column was used as stationary phase,and a mixed solution of acetonitrile and acetate buffer solution（mixed in the ratio of 6 to 94） was used as mobile phase.The eluate from the column was used for HPLC analysis with fluorescence detector.Linear relationship between the values of peak area and mass concentration of MG was obtained in the range of 1.0-500.0 μg·L-1,with detection limit（3S/N） of 0.46 μg·kg-1.Values of recovery and RSD′s（n=6） of the method found were in ranges of 78.0%-95.2% and 3.01%-5.98%.It was found that the transformation rate of MG into LMG was in the range of 99.1%-100.8%.