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基于UPLC-MS/MS的2型糖尿病患者血清胆汁酸代谢产物分析
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陈冰婷,伊丽则热·艾拜杜拉,李德龙,马晓丽 新疆医科大学药学院 
基金项目:新疆维吾尔自治区高校科研计划(XJEDU2019Y026);新疆维吾尔自治区自然科学基金(2018D01C170)
中文摘要:应用基于超高效液相色谱-串联质谱(UPLC-MS/MS)技术的代谢组学方法检测2型糖尿病患者血清胆汁酸代谢谱,获得代谢标志物,发掘可预测糖尿病的潜在预防治疗靶点。采用Waters ACQUITY UPLC BEH C18色谱柱(150 mm×2.1 mm,1.7 μm)分离,0.05%乙酸水溶液-乙腈为流动相,以0.40 mL/min流速进行梯度洗脱,用此方法测定了30例糖尿病患者和 50例正常人血清胆汁酸代谢谱。41种胆汁酸标准品在30 min内均得到良好分离,各成分在一定浓度范围内呈良好线性关系,检出限(LOD)为0.24~7.81 nmol/L,定量下限(LOQ)为0.49~15.63 nmol/L,相关系数(r2)为 0.986 7~0.999 7。血清中胆汁酸代谢谱与两组间存在相关性,通过正交偏最小二乘法-判别分析(OPLS-DA)发现,有12种胆汁酸(包括甘氨熊脱氧胆酸、甘氨胆酸、牛磺鹅脱氧胆酸、胆酸、熊去氧胆酸、甘氨鹅脱氧胆酸、甘氨脱氧胆酸、12 酮基石胆酸、鹅脱氧胆酸、脱氧胆酸、异石胆酸、石胆酸)承载了分组的重要信息。结合受试者工作特征(ROC)分析,最终获得牛磺鹅脱氧胆酸(AUC=0.68)、甘氨鹅脱氧胆酸(AUC=0.66)、甘氨熊脱氧胆酸(AUC=0.63)、胆酸(AUC=0.63)、脱氧胆酸(AUC=0.63)、甘氨脱氧胆酸(AUC=0.58) 6种胆汁酸代谢标志物,对于糖尿病的早期诊断及深入研究具有潜在的科研及临床价值。
中文关键词:超高效液相色谱-串联质谱(UPLC-MS/MS)  胆汁酸代谢  2型糖尿病  血清
 
Analysis of Bile Acid Metabolites in Serum of Type 2 Diabetic Patients Based on UPLC-MS/MS
Abstract:In this research,a metabolomics method of ultra high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) was developed for the detection of bile acid metabolites in serum of type 2 diabetic patients,obtaining metabolic markers and exploring potential preventive and therapeutic targets that may predict diabetes.The samples were separated on a Waters ACQUITY UPLC BEH C18 column(150 mm×2.1 mm,1.7 μm) by gradient elution with 0.05% acetic acid aqueous solution and acetonitrile as mobile phases at a flow rate of 0.40 mL/min.And then this method was used to determine the bile acid metabolites in serum of 30 diabetic patients and 50 normal persons.41 bile acid standard solutions were well isolated within 30 min.There were good linear relationships for 41 bile acids in certain concentration ranges,with their correlation coefficients(r2) of 0.986 7-0.999 7.The limits of detection(LOD) and the limits of quantitation(LOQ) were in the ranges of 0.24-7.81 nmol/L and 0.49-15.63 nmol/L,respectively.Results showed that there was a correlation between the bile acid metabolic spectra of the serum and the two groups.Through the orthogonal projections to latent structures-discriminant analysis(OPLS-DA),there are 12 kinds of bile acids,including glycoursodeoxycholic acid,glycocholic acid,taurochenodeoxycholic acid,cholic acid,ursodeoxycholic acid,glycochenodeoxycholic acid,glycodeoxycholic acid,12 ketolithocholic acid,chenodeoxycholic acid,deoxycholic acid,isolithocholic acid and lithocholic acid carrying a group of important information.Combined with the receiver operating characteristic curve(ROC curve),six bile acid metabolic markers,including taurochenodeoxycholic acid(AUC=0.68),glycochenodeoxycholic acid(AUC=0.66),glycoursodeoxycholic acid(AUC=0.63),cholic acid(AUC=0.63),deoxycholic acid(AUC=0.63) and glycodeoxycholic acid(AUC=0.58) were finally obtained,which have a potential value for the early diagnosis and further research of diabetes.
Key Words:ultra-high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)  bile acid metabolism  type 2 diabetes mellitus  serum
引用本文:陈冰婷,伊丽则热·艾拜杜拉,李德龙,马晓丽.基于UPLC-MS/MS的2型糖尿病患者血清胆汁酸代谢产物分析[J].分析测试学报,2020,39(12):1451-1457.
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