孕妇尿中五种邻苯二甲酸酯代谢产物的液-液萃取反相高效液相色谱测定法

Determination of Five Phthalate Metabolites in Urine of Pregnant Women by Liquid-Liquid Extraction-Reversed-Phase High Performance Liquid Chromatography

目的建立孕妇尿液中的邻苯二甲酸酯(PAEs)代谢产物的反相高效液相色谱(HPLC)测定方法。方法萃取剂正己烷用量为10 ml,萃取3次。采用GP-Phenyl 120A型色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈(0.2%乙酸)-水(0.2%乙酸)=45∶55(V/V),检测波长为228 nm,流量为1.0 ml/min。采用液-液萃取反相高效液相色谱法对孕妇尿液中五种PAEs代谢产物[邻苯二甲酸单甲酯(MMP)、邻苯二甲酸单乙酯(MEP)、邻苯二甲酸单丁酯(MBP)、邻苯二甲酸单丁苄酯(MBzP)、邻苯二甲酸-单-乙基己基酯(MEHP)]进行测定。结果在50～10 000μg/L的线性范围内,所得MMP、MEP、MBP、MBzP、MEHP的回归方程均呈较好的线性关系,相关系数≥0.99,检出限分别为4.17、4.31、12.74、10.66、13.12μg/L,平均回收率在80.99%～90.74%之间,RSD在1.580%～4.999%之间。结论该方法操作简便、回收率高、重现性好、检出限低,适用于尿液中5种邻苯二甲酸单酯的同时测定。

Objective To establish a method for the determination of phthalate metabolites in urine by reversed-phase high-performance liquid chromatography. Methods Phthalate metabolites were extracted by using liquid-liquid extraction. The optimal conditions of liquid-liquid extraction were that the usage amount of n-hexane was 10 ml for three times. The wave-length of determine was 228 nm. The contents of 5 kinds of phthalates metabolites （MMP,MEP,MBP,MBzP and MEHP） were determined by using RT-HPLC （GP-Phenyl 120A 4.6 mm×250 mm,5 μm）; methyl cyanides （0.2% acetic acid）：water （0.2% acetic acid）=45：55 （V：V）. Results The correlation coefficients were ≥0.99. The limits of detection were 4.17,4.31,12.74, 10.66 and 13.12 μg/L. The average recovery rates were 80.99%-90.74% and the relative standard deviations were 1.580%-4.999%. Conclusion This method has good recovery rate,reproducibility,lower limit of detection and can be used for the determination of five kinds of phthalate metabolites in urine.