表面活性剂辅助微萃取-高效液相色谱法测定尿中一氯苯的2种代谢物

HPLC Determination of 2 Metabolites of Monochlorobenzene in Urine with Surfactant Assisted Microextraction

尿样以十六烷基三甲基溴化铵为辅助剂,正辛醇为萃取剂,萃取液离心后,取上层有机相,采用高效液相色谱法测定其中一氯苯的2种代谢物(4-氯邻苯二酚和对氯苯酚)的含量。以RD-C18色谱柱为分离柱,以甲醇与0.15%(体积分数)的磷酸溶液以体积比65∶35组成的混合液为流动相,在检测波长280nm处进行测定。4-氯邻苯二酚和对氯苯酚的质量浓度均在600μg·L^-1以内与其对应的峰面积呈线性关系,检出限(3S/N)为1.67,2.23μg·L^-1,测定下限(10S/N)为5.55,7.43μg·L^-1。以空白样品为基体进行加标回收试验,所得回收率为89.2%~97.4%,测定值的日内相对标准偏差(n=6)为2.1%~4.9%,日间相对标准偏差(n=18)为2.5%~5.3%。

The urine sample was extracted with n-octanol with hexadecyl trimethyl ammonium bromide as auxiliary solvent.The extract was centrifugated,and then the upper organic phase was taken.HPLC was applied to the determination of 2 metabolites(4-chlorocatechol and p-chlorophenol)of monochlorobenzene in the upper organic phase.RD-C18 chromatographic column was used as separation column,and a mixture of methanol and 0.15%(φ)phosphoric acid solution with volume ratio of 65∶35 was used as mobile phase.Detection at 280 nm was adopted in the determination.Linear relationships between values of peak area and mass concentration of 4-chlorocatechol and p-chlorophenol were kept in the same range within 600μg·L^-1,with detection limits(3 S/N)of 1.67,2.23μg·L^-1 and lower limits of determination(10 S/N)of 5.55,7.43μg·L^-1 respectively.On the base of blank sample,test for recovery was made by standard addition method;values of recovery found were in the range of 89.2%-97.4%,with intra-day RSDs(n=6)and inter-day RSDs(n=18)of determined values in the ranges of 2.1%-4.9%,2.5%-5.3%,respectively.