| Objectives Hydroxylated polycyclic aromatic hydrocarbons(OH-PAHs),as metabolites of polycyclic aromatic hydrocarbons(PAHs)in urine,are commonly used as internal biomarkers to comprehensively estimate the total external PAHs exposure in humans and environmental risk.The accurate and reliable determination of OH-PAHs in urine is usually difficult due to their trace concentration and the complexity of biological matrices complicated matrix.Consequently,sample pretreatment approaches should be employed before chromatographic determination.In this manuscript,a solid supported liquid-liquid extraction(SLE)coupled with ultrahigh performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method and was developed for the determination of seven hydroxyl polycyclic aromatic hydrocarbons(OH-PAHs)in urine samples,and graphene oxide(GO)bonded diatomite(Dt)composites were prepared and used as solid phase extraction sorbent for four OH-PAHs in urine samples using high performance liquid chromatography fluorescence detector(HPLC-FLD).Methods 1 The SLE was used as the sample pretreatment method for simultaneous detection of seven kinds of OH-PAHs in urine samples and the determination was carried out by UPLC-MS/MS,SLE parameters including eluting solvent and volume,and the nitrogen evaporation pressure were investigated.Under the optimized condition,the method was evaluated in terms of linearity,precision,recovery,limits of detection(LODs),and limits of quantification(LOQs).The feasibility of this method was further performed by the determination of seven OH-PAHs in urine samples of petroleum workers and control.2 GO was covalently bonded to the amino Dt particles using N,N’-dicyclohexylcarbodiimide as coupling agent to form Dt-NH2@GO composites by coupling the amino groups of amino Dt and the carboxyl groups of GO.The effects of the amount of GO(0-10 mg)and the coating times(1-2 times)on the adsorption efficiency were evaluated using four OH-PAHs.The prepared Dt-NH2@GO was characterized by scanning electron microscopy(SEM),fourier-transform infrared spectroscopy(FT-IR)and elemental analysis.The extraction parameters including loading volume,eluting solvent and volume were optimized.Finally,linearity,limit of quantization,limit of detection(LOD),limit of quantification(LOQ),recovery,precision and stability of the method was validated.Furthermore,the method was performed by the determination of four OH-PAHs in urine samples of smokers.Results 1 The enzymatic hydrolyzed urine(0.8 mL)was loaded to a SLE cartridge,and then the adsorbed analytes were eluted by 6 mL of ethyl acetate,and the collected eluate was evaporated under nitrogen at a pressure of 48 kPa at 25°C until dryness.Under the optimized conditions,the developed method gave the detection limits(S/N=3)of 0.05-1.0 ng/mL,the quantitation limits(S/N=10)of 0.15-3.0 ng/mL,and the linear range of 0.3-3000 ng/mL(r ≥ 0.9951).The recoveries for spiked OH-PAHs in urine samples were in the range of 74.8%-117%,and the intra-day(n=3)and inter-day(n=3)precisions were 3.7%-9.6% and 3.7%-10.8%,respectively.The proposed method was successfully applied in the determination of OH-PAHs in urine from several petroleum workers and control group.Except 2-hydroxy-phenanthrene and 9-hydroxy-phenanthrene,the concentrations of the other six OH-PAHs in the urine samples of petroleum workers were much higher than that in control samples(p < 0.05).According to the results of SEM,FT-IR and elemental analysis,GO was successfully bonded to the surface of the amino Dt particles.One hundred milligram Dt-NH2@GO composites was used as solid phase extraction sorbent for the determination of four OH-PAHs in urine samples.The SPE process was performed on the Dt-NH2@GO composites loading 3 mL enzymatic hydrolyzed urine,and 3 mL ACN as eluting solvent.The linearity of the developed method was 0.1-200 ng/mL for four OHPAHs(r ≥ 0.9995).The detection limits(S/N=3)and the quantitation limits(S/N=10)of four OH-PAHs were 0.04-0.15 ng/mL and 0.1-0.4 ng/mL,respectively.The average recoveries at three spiked levels of four OH-PAHs were 90.6%-100%,with the intra-day(n=3)and inter-day(n=3)relative standard deviations(RSDs)were 1.8%-6.4% and 2.7%-11.8%,respectively.Further,the intra batch(n=3)and inter batch(n=3)RSDs were no more than 14.5%.Conclusions 1 A SLE coupled with UPLC-MS/MS method was developed for the determination of seven OH-PAHs in urine samples,the method can be applied to for the facile and efficient monitoring of OH-PAHs from urine samples.2 The Dt-NH2@GO composites were developed as a low-cost SPE sorbent coupled with UPLC-MS/MS for the rapid and accurate determination of OH-PAHs in urine.Figure 14;Table 10;Reference 152... |
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