人尿中9种单羟基多环芳烃的固相萃取-超高效液相色谱-串联质谱同时测定法

Simultaneous determination of nine monohydroxypolycyclic aromatic hydrocarbons in urine by using solid phase extraction-ultra performance liquid chromatography tandem mass spectrometry

目的建立同时测定尿中9种单羟基多环芳烃(OH-PAHs,即1-羟基萘、2-羟基萘、3-羟基芴、3-羟基菲、1-羟基芘、3-羟基■、6-羟基■、3-羟基苯并蒽和9-羟基苯并芘)的固相萃取-超高效液相色谱-串联质谱(SPE-UPLC-MS/MS)法。方法将10 ml尿样经酶水解、C18固相萃取柱(500 mg,3 ml)提取富集后,用Waters ACQUITY UPLC BEH C_(18)色谱柱(2.1×100 mm,1.7μm)分离,在三重四级杆串联质谱负离子模式下检测,以内标法定量分析。结果尿中9种目标化合物在0.102~40 ng/ml范围内均呈良好的线性关系(r>0.999),检出限(S/N=3)为0.01~0.32 ng/ml,样品在1.6、4.0、10 ng/ml浓度水平的平均加标回收率为91.5%~128%,相对标准偏差(RSD)<15%。结论该方法用时短,灵敏度高,准确度好,精密度高,适合人群尿中多环芳烃代谢物的监测。

Objective To establish a method for the simultaneous determination of nine monohydroxypolycyclic aromatic hydrocarbons （OH-PAHs） in urine by using SPE-UPLC-MS/MS. Methods An aliquot of 10 ml urine was hydrolyzed with enzyme and extracted by Ci8 SPE cartridge （500 mg, 3 ml）. The extract was separated on Waters ACQUITYUPLC BEH C18 column （2.1×100 mm, 1.7 μm）. Then the target analytes, including 1-NAP, 2-NAP, 3-FLUO, 3-PHEN, 1-PYR, 3-CHRY, 6-CHRY, 3-BAA and 9-BAP, were detected in negative ionization mode of tandem mass spectrometry and at last quantified by internal standard method. The experimental parameters such as solid phase extraction column, elutiou solvent and its volume, analytical column, gradient conditions, and mass spectrometry parameters were optimized. Results The analytes had good linearity range （0.102-40 ng/ml, r〉0.999）. The limits of detection were 0.01-0.32 ng/ml. The average recoveries at three different concentration levels （1.6, 4.0, 10 ng/ml） were 91.5%-128%, and the inter-day RSDs were less than 15%. Conclusion The proposed method is time-saving,accurate, sensitive, as well as lower LOD, and it is applicable to the rapid analysis of OH- PAHs in urine.