人尿中4种有机磷酸酯阻燃剂代谢物的固相萃取-高效液相色谱-串联质谱测定法

Determination of four metabolites of organophosphate flame retardants in human urine by solidphase extraction and high-performance liquid chromatography-tandem mass spectrometry

目的建立人尿中磷酸二(1,3-二氯异丙基)酯(BDCPP)、磷酸二(2-氯丙基)酯(BCPP)、磷酸二甲苯酯(BCP)和磷酸二苯酯(DPHP)4种有机磷酸酯阻燃剂代谢物(DAPs)的固相萃取-高效液相色谱-串联质谱检测方法。方法取尿样2ml,加2 ml水稀释,以Strata TM-X-AW弱阴离子交换柱富集净化,微弱氮气吹干后,25%甲醇-水复溶至400μl,过0.22μm尼龙滤膜,1 mmol/L乙酸铵水溶液-甲醇为流动相,经DIKMA HPLC Spursil C18色谱柱(250 mm×2.1 mm,3μm)梯度洗脱分离,电喷雾离子源负离子模式(ESI-),多反应监测(MRM)模式检测。结果 DPHP、BDCPP、BCP在0.05~200μg/L、BCPP在0.1~250μg/L的线性范围内,各目标化合物的回归方程均呈良好的线性关系,r>0.999。该方法的检出限为0.01~0.4μg/L,定量下限为0.05~1.3μg/L;平均回收率为63.0%~109.4%,RSD为2.1%~13.1%。结论该方法操作简单,灵敏可靠,适用于人尿中4种有机磷酸酯代谢物的同时测定。

Objective To establish a method for the determination of(1,3-dichloroisopropyl)phosphate(BDCPP),di(2-chloropropyl)phosphate(BCPP),dibenzyl phosphate(BCP)and triphenyl phosphate(DPHP)in human urine by solid-phase extraction(SPE)with high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods The urine samples(2 ml)were diluted by 2 ml water,purified through StrataTM-X-AW weak anion solid phase extraction cartridge,concentrated to dryness under weak nitrogen,reconstituted to 400μl with 25%methanol-water and filtered with 0.22μm nylon filter.The four metabolites of organophosphate flame retardants were separated by DIKMA HPLC Spursil C18 column(250 mm×2.1 mm,3μm)with methanol and 1 mmol/L ammonium acetate aqueous solution as mobile phase for gradient elution,then determined by HPLC-MS/MS under multiple reaction monitoring with negative ion mode electrospray ionization.Results All calibration curves except BCPP(showed good linearity in 0.10-250μg/L)showed good linear relationship in the range from 0.05μg/L to 200μg/L with good correlation coefficients(r>0.999).The limits of detection were 0.01-0.4μg/L and the limits of quantification were 0.05-1.3μg/L.The average spiked recoveries for four targets were 63.0%-109.4%with RSDs of 2.1%-13.1%.Conclusion The method is simple,sensitive,reliable and is applicable to simultaneous determination of the four metabolites of organophosphate flame retardants in human urine in the study.