尿液中典型胆汁酸的超高效液相色谱-串联三重四级杆质谱分析方法  

Analysis of typical bile acids in urine by ultra-performance liquid chromatography coupled to triple quadrupole mass spectrometry

作　　者：胡立成[1] 朱传新[2] 

机构地区：[1]乐清市疾病预防控制中心,浙江温州325600 [2]温州市疾病预防控制中心,浙江温州325001

出　　处：《中国卫生检验杂志》2017年第24期3523-3525,3528,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的建立尿中胆酸、脱氧胆酸、鹅脱氧胆酸和石胆酸的超高效液相色谱-串联三重四级杆质谱同时分析方法。方法取1.0 ml尿样,用β-葡萄糖醛酸酶水解后,使用混合型强阴离子交换固相萃取技术净化,反相超高效液相色谱柱和含0.1%甲酸的甲醇-水梯度洗脱分离后,采用阴离子模式下的电喷雾三重四极杆质谱检测,内标标准曲线法定量。结果 4种胆汁酸检出限在0.2 ng/ml^1.2 ng/ml。胆酸、脱氧胆酸和石胆酸线性范围为1.0 ng/ml^500 ng/ml,而鹅脱氧胆酸线性范围为5.0 ng/ml^500 ng/ml;4种胆汁酸线性相关系数为0.991~0.998,尿样平均加标回收率率为83.7%~109.7%,相对标准偏差为5.4%~13.8%。20名成人尿样中胆酸、脱氧胆酸、鹅脱氧胆酸和石胆酸均被检出,浓度分别为(164.1±87.6)ng/ml、(156.7±145.3)ng/ml、(165.8±106.1)ng/ml和(6.5±6.1)ng/ml。结论该方法快速、灵敏、准确,完全满足尿中典型胆汁酸的分析要求。Objective To develop an analytical method for simultaneous determination of cholic acid,deoxycholic acid,chenodeoxycholic acid and lithocholic acid in urine by ultra-performance liquid chromatography coupled to triple quadrupole mass spectrometry（ UPLC-MS/MS）. Methods After 1. 0 ml of urine was hydrolyzed by β-glucuronidase and purified by mixed-mode anion exchange solid phase extraction,the bile acids were separated with the gradient elution of mobile phase of methanol and water containing 0. 1% formic acid and detected by triple quadrupole mass spectrometry with the electrospray ion source in negative ion mode. The quantification was based on internal standard calibration curve method. Results The detection limits of the four bile acids varied within 0. 2 ng/ml-1. 2 ng/ml. The linear ranges of cholic acid,deoxycholic acid,and lithocholic acid were within 1. 0 ng/ml-500 ng/ml and that of chenodeoxycholic acid was within 5. 0 ng/ml-500 ng/ml. The linear correlation coefficients of four bile acids varied within 0. 991-0. 998. The average recoveries of bile acids spiked in real urine samples varied within 83. 7%-109. 7%,with relative standard deviations within 5. 4%-13. 8%. Four bile acids were detected in 20 real adult urine samples and the concentrations ± standard deviations of cholic acid,deoxycholic acid,chenodeoxycholic acid,and lithocholic acid were（ 164. 1 ± 87. 6） ng/ml,（ 156. 7 ± 145. 3） ng/ml,（ 165. 8 ± 106. 1） ng/ml,and（ 6. 5 ± 6. 1） ng/ml,respectively. Conclusion The method is rapid,sensitive,accurate and suitable for the determination of bile acids in urine.

关 键 词：尿液 胆汁酸 超高效液相色谱-串联质谱法 

分 类 号：O657.63[理学—分析化学]