固相萃取-超高效液相色谱/串联质谱法检测人体血清中邻苯二甲酸二(2-乙基已基)酯代谢物

建立了固相萃取-超高效液相色谱/串联质谱法检测人体血清中邻苯二甲酸二(2-乙基已基)酯(DEHP)代谢物.血清样品中加入乙酸钠溶液及内标13C4邻苯二甲酸单乙基已基酯(13C4MEHP)后,在37℃条件下用β-葡萄糖醛苷酶酶解提取12h,提取液经MAX固相萃取小柱净化并浓缩到纯水中.以甲醇5 mmol/L乙酸铵溶液为流动相,经Waters UPLC(R)HSS T3色谱柱分离后,在电喷雾离子源负离子模式下,以多反应监测(MRM)方式进行扫描,内标法定量.DEHP代谢物在1.0～100.0 μg/L范围内线性关系良好,检出限均为0.1 μg/kg,回收率89.6％～103.2％.该方法灵敏度高,准确度好,可作为监测人群内暴露的重要技术支撑.

Determination of DEHP Metabolites in Human Serum by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry with Solid Phase Extraction

A sensitive and accurate analytical method was developed and validated for the determination of di-(2-ethylhexyl)-phthalate(DEHP) metabolites in human serum by ultra performance liquid chromatography tandem mass spectrometry with solid phase extraction.The serum added with sodium acetate solution and internal standard (13C4MEHP) was hydrolyzed with beta-Glucuronidaseat at 37 ℃ for 12 hours.The pretreated sample was condensed in the purified water after loading,washing and purifying through MAX solid phase extraction column.DEHP metabolites were separated and quantitated by an UPLC-MS/MS method with solid phase extraction.The separation was achieved using a Waters UPLC(R) HSS T3 column and a binary gradient mobile phase of methanol-5 mmol/L ammonium acetate solution.A triple quadrupole mass spectrometer with ESI negative mode was used as a detector.The linear range of calibration curves of DEHP metabolites was 1.0-100.0 μg/L.The recoveries(n=6) were in the range of 89.6％-103.2％.This highly sensitive and accurate method can be used as an important technical support tool to monitor the population exposed to DEHP.