固相萃取-高效液相色谱法测定鱼制品中8种杂环胺类化合物

HPLC Determination of 8 Heterocyclic Amines in Fish Products with Solid Phase Extraction

鱼制品样品(2g)用2.0mol·L-1氢氧化钠溶液10mL超声提取20min,用二氯甲烷50mL淋洗装有15g硅藻土的层析柱,洗脱液经MCX柱进行固相萃取分离。用氨水-甲醇(10+90)混合液淋洗MCX柱,洗脱液于40℃吹氮蒸干,残渣用甲醇1.0mL溶解供色谱分析。以TOSOH TSK gel ODS-80TM色谱柱为固定相,用(A)乙腈和(B)pH 3.5的0.01mol·L-1磷酸溶液以不同体积比组成的混合液作流动相进行梯度淋洗。用二极管阵列检测器测定。8种杂环胺类化合物的质量浓度均在0.005~1.0mg·L-1范围内与其峰面积呈线性关系。方法的检出限(3S/N)在0.3~3.8μg·kg-1之间。以空白样品为基体进行加标回收试验,所得回收率在51.8%~93.7%之间,测定值的相对标准偏差(n=6)在0.89%~3.7%之间。

Sample of fish product(2g)was alkalified with 10mL of 2.0mol·L-1 NaOH solution and extracted ultrasonically for 20min.50mL of CH2Cl2 were used to elute the column equipped with 15g of diatomite.The eluate was passed through a MCX column for SPE separation.Mixture of NH3·H2O-CH3OH(10+90)were to elute the MCX column.The eluate was evaporated to dryness by N2-blowing at 40℃,and the residue dissolved in 1.0mL of CH3OH for HPLC analysis.TOSOH TSK gel ODS-80TM column was used for chromatographic separation,and mixtures of (A)CH3CN and(B)0.01mol·L-1 H3PO4solution pH 3.5mixed in different volumic ratio were used as mobile phase in gradient elution.PAD were used in the determination.Linear relationships between values of peak area and mass concentration of the 8heterocyclic amines were kept in the same range of 0.005-1.0mg·L-1,with detection limits(3S/N)in the range of 0.3-3.8μg·kg-1.On the base of blank sample,test for recovery was made by standard addition method;values of recovery found were in the range of 51.8%-93.7%,with RSD′s(n=6)in the range of 0.89%-3.7%.