固相支撑液液萃取结合超高效液相色谱-串联质谱法同时测定尿液中7种多环芳烃代谢物
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摘要
建立了固相支撑液液萃取法(SLE)结合超高效液相色谱-串联质谱法(UPLC-MS/MS)测定尿液中1-羟基萘、2-羟基萘、2-羟基芴、9-羟基芴、2-羟基菲、9-羟基菲和1-羟基芘7种羟基多环芳烃(OH-PAHs)的方法。实验将0.8 mL尿液酶解液经SLE萃取,考察了洗脱溶剂种类及体积的影响,并对氮吹浓缩压力进行了优化。尿液样品使用6 mL乙酸乙酯洗脱,洗脱液在48 kPa(25℃)压力下氮吹至干后,经UPLC-MS/MS对7种OH-PAHs进行分析,采用同位素内标法定量。方法在0.3～3 000 mg·L~(-1)范围内线性关系良好(r≥0.995 1),检出限和定量下限分别为0.05～1.0 mg·L~(-1)和0.15～3.0 mg·L~(-1)。加标3个浓度水平的回收率为70.8%～117%,日内(n=3)和日间(n=3)相对标准偏差分别为3.7%～9.6%和3.7%～11%。采用本方法对10例石油工人和10例对照人群(在校大学生)尿液中的OH-PAHs水平进行分析,除2-羟基菲和9-羟基菲外,其余5种OH-PAHs在石油工人尿液中的含量均高于对照人群(p<0.05),差异具有统计学意义。该方法经上样和洗脱两步完成样品前处理,简单高效,可用于尿液样品中OH-PAHs的分析。
An ultrahigh performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) with solid supported liquid-liquid extraction(SLE) was developed for the determination of seven hydroxyl polycyclic aromatic hydrocarbons(OH-PAHs) in urine samples.0.8 mL enzymatic hydrolyzed urine sample was extracted on a SLE cartridge.Parameters for SLE,including type and volume of eluting solvent,and nitrogen evaporation pressure were optimized.The adsorbed analytes were eluted with 6 mL ethyl acetate,and the collected eluate was evaporated to dryness under nitrogen at a pressure of 48 kPa at 25 ℃,then analyzed by UPLC-MS/MS and quantified by the internal standard method.Under the optimized conditions,the calibration curves for the analytes were linear in the range of 0.3-3 000 mg·L~(-1),with correlation coefficients not less than 0.995 1.The detection limits(S/N=3) and quantitation limits(S/N=10) were in the ranges of 0.05-1.0 mg·L~(-1) and 0.15-3.0 mg·L~(-1),respectively.The recoveries for OH-PAHs in urine samples at three spiked levels ranged from 70.8% to 117%,with the intra-day(n=3) and inter-day(n=3) RSDs of 3.7%-9.6% and 3.7%-11%,respectively.The proposed method was successfully applied in the determination of OH-PAHs in urine from 10 petroleum workers and 10 control groups(college students).Except 2-hydroxy-phenanthrene and 9-hydroxy-phenanthrene,the concentrations of the other five OH-PAHs in the urine samples of petroleum workers were much higher than that in the control samples(p<0.05).The proposed SLE method only included two steps,therefore,the proposed method is applicable for the facile and efficient monitoring of OH-PAHs in urine samples.
An ultrahigh performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) with solid supported liquid-liquid extraction(SLE) was developed for the determination of seven hydroxyl polycyclic aromatic hydrocarbons(OH-PAHs) in urine samples.0.8 mL enzymatic hydrolyzed urine sample was extracted on a SLE cartridge.Parameters for SLE,including type and volume of eluting solvent,and nitrogen evaporation pressure were optimized.The adsorbed analytes were eluted with 6 mL ethyl acetate,and the collected eluate was evaporated to dryness under nitrogen at a pressure of 48 kPa at 25 ℃,then analyzed by UPLC-MS/MS and quantified by the internal standard method.Under the optimized conditions,the calibration curves for the analytes were linear in the range of 0.3-3 000 mg·L~(-1),with correlation coefficients not less than 0.995 1.The detection limits(S/N=3) and quantitation limits(S/N=10) were in the ranges of 0.05-1.0 mg·L~(-1) and 0.15-3.0 mg·L~(-1),respectively.The recoveries for OH-PAHs in urine samples at three spiked levels ranged from 70.8% to 117%,with the intra-day(n=3) and inter-day(n=3) RSDs of 3.7%-9.6% and 3.7%-11%,respectively.The proposed method was successfully applied in the determination of OH-PAHs in urine from 10 petroleum workers and 10 control groups(college students).Except 2-hydroxy-phenanthrene and 9-hydroxy-phenanthrene,the concentrations of the other five OH-PAHs in the urine samples of petroleum workers were much higher than that in the control samples(p<0.05).The proposed SLE method only included two steps,therefore,the proposed method is applicable for the facile and efficient monitoring of OH-PAHs in urine samples.
引文
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[3]Kim K H,Jahan S A,Kabir E,Brown R J.Environ.Int.,2013,60(5):71-80.
[4]Lu C D,Zhang F.Chin.Occup.Med.(卢城德,张放.中国职业医学),2017,44(4):502-504.
[5]Wang D J,Sun Z T,Yang C,Wang B L,Pang S T,Pang Z C.Chin.J.Public Health(王德杰,孙治涛,杨超,王炳玲,逄淑涛,逄增昌.中国公共卫生),2015,31(8):1100-1105.
[6]Shrivastava M,Lou S,Zelenyuk A,Easter R C,Corley R A,Thrall B D,Rasch P J,Fast J D,Massey Simonich SL,Shen H,Tao S.Proc.Natl.Acad.Sci.USA,2017,114(6):1246-1251.
[7]Large C,Wei Y D.Environ.Sci.Pollut.Res.,2017,24(21):17874-17880.
[8]Scherer G,Frank S,Riedel K,Meger-Kossien I,Renner T.Cancer Epidemiol.Biomarkers Prevent.,2000,9(4):373-380.
[9]Wang Y S,Meng L,Pittman E N,Etheredge A,Hubbard K,Trinidad D A,Kato K,Ye X Y,Calafat A M.Anal.Bioanal.Chem.,2017,409(4):1-7.
[10]Shen M,Xing J,Ji Q,Li Z,Wang Y,Zhao H,Wang Q,Wang T,Yu L,Zhang X,Sun Y,Zhang Z,Niu Y,Wang H,Chen W,Dai Y,Su W,Duan H.Environ.Sci.Technol.,2018,52(11):6610-6616.
[11]Bal C,Aˇgi爧E R,Büyük爧ekerci M,Gündüz9z M,Tutkun L,YilmazH.Am.J.Ind.Med.,2018,61(6):471-476.
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[13]You F,Zhu L,He L,Ran L J,Jin Y,Sun C J.Chin.J.Anal.Chem.(游钒,朱岚,何玲,冉良骥,金燕,孙成均.分析化学),2014,42(12):1723-1728.
[14]Li P,Cui J T,Guo X X,Zeng X Y,Yu Z Q.J.Instrum.Anal.(李佩,崔君涛,郭溪香,曾祥英,于志强.分析测试学报),2018,37(3):282-287.
[15]Zhou L J,Hu Y L,Li G K.Anal.Chem.,2016,88:6930-6938.
[16]Gaudreau,BérubéR,Bienvenu J F,Fleury N.Anal.Bioanal.Chem.,2016,408(15):4021-4033.
[17]Zhang H F,Xu H.J.Chromatogr.A,2017,1521:27-35.
[18]Yang B C,Fang S F,Wan X J,Luo Y,Zhou J Y,Li Y,Li Y J,Wang F,Huang O P.Anal.Chim.Acta,2017,973:68-74.
[19]Fan R F,Ramage R,Wang D L,Zhou J Q,She J W.Talanta,2012,93(11):383-391.
[20]Chauhan A,Bhatia T,Singh A,Saxena P N,Kesavchandran C,Mudiam M K.J.Chromatogr.B,2015,985:110-118.
[21]Ding C M,Jin Y L,Lin S B.Chin.J.Anal.Chem.(丁昌明,金银龙,林少彬.分析化学),2012,40(3):397-402.
[22]Liu H,Huang L,Chen Y,Guo L,Li L,Zhou H,Luan T.J.Chromatogr.B,2015,992:96-102.
[23]GAO X,Tan Y L,Guo H.J.Chromatogr.B,2017,1052:27-33.
[24]Seo J E,Park J E,Lee J Y,Kwon H.Food Anal.Methods,2016,9:1595-1605.
[25]Gao X,Guo H,Zhang W W,Gu C.J.Anal.Toxicol,2016,40(6):437-444.
[26]Wang L,Li L,Zhou J P,Xu Q J.Mod.Prevent.Med.(王丽,李磊,周靖平,许秋瑾.现代预防医学),2012,39(23):6238-6241.