超高效液相-串联质谱分析法同时测定蕲蛇提取物中核苷及氨基酸的含量  

Simultaneous determination of nucleosides and amino acids in extract of Agkistrodon acutus by ultra-high performance liquid chromatography-tandem mass spectrometry

作　　者：王蓉蓉 胡争艳[2] 吴平谷[2] 单乐天[1] 黄家卫[1] 王春雷[3] WANG Rong-rong;HU Zheng-yan;WU Ping-gu;SHAN Le-tian;HUANG Jia-wei;WANG Chun-lei(Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310053,China)

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]浙江省疾病预防控制中心,浙江杭州310051 [3]浙江省肿瘤医院,浙江杭州310022

出　　处：《中国卫生检验杂志》2020年第15期1793-1797,1803,共6页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金项目(81774331);浙江省中医药科技计划项目(2016ZQ010)。

摘　　要：目的建立蕲蛇提取物中4种核苷(尿苷、尿嘧啶、黄嘌呤、次黄嘌呤)及4种氨基酸(L-羟脯氨酸、L-天冬氨酸、L-谷氨酸、甘氨酸)的超高效液相-串联质谱分析法(UPLC-MS/MS),并对蕲蛇酒浸提取物与高温水提物中核苷及氨基酸类成分的含量进行对比分析研究。方法蕲蛇提取物经甲醇-水(1∶1,V/V)超声溶解后采用HSS T3色谱柱(100 mm×2.1 mm,1.8μm)分离,以0.1%甲酸水和乙腈为流动相进行梯度洗脱,采用电喷雾正离子模式(ESI+)在多反应监测(MRM)模式进行监测,外标法定量。结果8种化合物线性关系良好,相关系数(r)均>0.995,各化合物的检出限和定量限分别为0.20 ng/ml^20.0 ng/ml和1.00 ng/ml^50.0 ng/ml,平均加标回收率为92.8%~102.4%,RSD为0.3%~5.2%。结论该方法操作简单、快速、准确,可用于蕲蛇提取物中多种核苷和氨基酸的分析。Objective A quick confirmative method was developed for the determination of 4 kinds of nucleosides(uridine,uracil,2,6-dihydroxypurine,6-hydroxypurine)and 4 kinds of amino acids(L-hydroxyproline,L-glutamic acid,L-aspartic acid,glycine)in extractant of Agkistrodon acutus by ultra-high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).The nucleosides and amino acids in the ethanol extracts and high temperature water extracts of Agkistrodon acutus were analyzed and compared.Methods The extracts of Agkistrodon acutus were dissolved in methanol-water(1∶1,V/V)and then analyzed on HSS T3 column(100 mm×2.1 mm,1.8μm)using 0.1%formic acid solution and acetonitrile as the mobile phase with gradient elution.The samples were determined by tandem mass spectrometry in the positive ion mode(ESI+)with the multiple reaction monitoring(MRM)mode.The quantitative analysis was performed by the external standard method.Results It was showed that 8 kinds of nucleosides and amino acids obtained good linearities with correlation coefficients(r)more than 0.995.The limits of detection(LOD)and quantitation(LOQ)were respectively within 0.20 ng/ml-20.0 ng/ml and 1.00 ng/ml-50.0 ng/ml.The recoveries of the 8 compounds were within 92.8%-102.4%and the relative standard deviations were within 0.3%-5.2%.Conclusion The developed method is simple,rapid,and accurate,which could simultaneously analyze multi-kinds of nucleosides and amino acids in Agkistrodon acutus extracts.

关 键 词：蕲蛇提取物 超高效液相-串联质谱分析法 核苷 氨基酸 

分 类 号：O657.63[理学—分析化学]