血清中胆汁酸的高效液相色谱荧光测定及质谱鉴定结果分析

Analysis of results of high-performance liquid chromatography fluorescence determination and mass spectrometry identification of bile acids in serum

目的应用超快速液相色谱联合质谱分析法对胆石症患者胆汁酸成分及其意义进行分析。方法建立超快速液相色谱联合质谱(HPLC-MS)测定血清中胆汁酸的方法。选取广东附属医院2018年10月~2019年4月收治的胆石症患者80例为研究对象,设为试验组,对照组受试者为健康人群,试验组受试者为胆石症患者,分为胆囊结石和胆管结石两种疾病。采集受试者血清样本,测定脂质代谢水平[胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(H-LDL)含量];应用HPLC-MS法测定两组受试者血清胆汁酸中6种亚组分[牛磺胆酸(TCA)、甘氨胆酸(GCA)、牛磺鹅脱氧胆酸(TDCA)、甘氨鹅脱氧胆酸(GCDCA)、甘氨脱氧胆酸(GDCA)、牛磺石胆酸(TLCA)]组成和含量的差别。结果对照组受试者中检测出了4种亚组分、试验组受试者中检测出了6种亚组分。试验组与对照组相比,TCA、GCA成分含量较高,差异有统计学意义(P<0.05)。TCA、GCA成分可能为胆石症疾病诊断的标志成分;试验组受试者中胆囊结石患者与胆管结石患者相比,GDCA成分含量不同,差异有统计学意义(P<0.05),该成分可能为胆石症不同发病部位的诊断成分;试验组与对照组相比,TC、TG、LDL脂质代谢指标均较高,差异有统计学意义(P<0.05),可作为胆石症疾病诊断的指标之一。结论通过建立一种HPLC-MS测定血清中胆汁酸的方法,为胆石症患者的胆石形成机制提供理论的依据,充实其形成的理论原因;找出胆石症患者的预测或治疗的检测标志物,可减少胆石症患者的发病率,节省社会资源,促进人们的健康。

Objective To analyze the bile acid composition in patients with gallstone disease by ultra-performance liquid chromatography coupled with mass spectrometry and its significance.Methods A method for determination of bile acids in serum by ultra-performance liquid chromatography coupled with mass spectrometry(HPLC-MS)was established.80 patients with gallstone disease admitted to Guangdong Affiliated Hospital from October 2018 to April 2019 were selected as the experimental group.The subjects in the control group were healthy people,and the subjects in the experimental group were patients with cholelithiasis which were further divided into gallbladder stones and bile duct stones.The serum samples were collected from subjects to determine lipid metabolism levels(total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),high density lipoprotein(H-LDL)levels).HPLC-MS method was used to determine the difference in composition and levels of 6 sub-components taurocholic acid(TCA),glycine cholic acid(GCA),taurodeoxycholic acid(TDCA),glycine chenodeoxycholic acid(GCDCA),glycine deoxycholic acid(GDCA),taurolithocholic acid(TLCA)in serum bile acids of the two groups.Results 4 sub-components were detected in the control group and 6 sub-components were detected in the experimental group.The levels of TCA and GCA in the experimental group was higher than those in the control group,and the differences were statistically significant(P<0.05).TCA and GCA components may be the hallmark components for the diagnosis of cholelithiasis.In the experimental group,the level of GDCA components between patients with gallbladder stones and patients with bile duct stones was different,and the difference was statistically significant(P<0.05).This component may be the diagnostic component of different locations of cholelithiasis.The TC,TG and LDL lipid metabolism indexes of the experimental group were higher than those of the control group,and the differences were statistically significant(P<0.05).All of them can be used as one of the indicators for the diagnosis of cholelithiasis.Conclusion By establishing a HPLC-MS method for the determination of bile acids in serum,we provide a theoretical basis for the gallstone formation in patients with cholelithiasis,and enrich the theoretical reasons for its formation.Finding out predictive or therapeutic markers for patients with gallstone disease can reduce the incidence of cholelithiasis,save social resources,and promote people’s health.