高效液相色谱-串联质谱法快速测定吸烟者尿液中8-羟基脱氧鸟苷和8-羟基鸟苷  

Rapid Determination of 8-Hydroxydeoxyguanosine and 8-Hydroxyguanosine in Urine of Smokers by HPLC-MS/MS

作　　者：吴凡 李惠杰 杨光宇 郭亚东 胡秋芬 梁梦洁 WU Fan;LI Huijie;YANG Guangyu;GUO Yadong;HU Qiufen;LIANG Mengjie(School of Ethnic Medicine,Yunnan Minzu University,Kunming650504,China;Technology Center,China Tobacco Yunnan Industrial Co.,Ltd.,Kunming650106,China;School of Pharmacy,Kunming Medical University,Kunming650504,China;Research and Development Center,Yunnan Yuntianhua Co.,Ltd.,Kunming650228,China)

机构地区：[1]云南民族大学民族医药学院,昆明650504 [2]云南中烟工业有限责任公司技术中心,昆明650106 [3]昆明医科大学药学院,昆明650504 [4]云南云天化股份有限公司研发中心,昆明650228

出　　处：《理化检验（化学分册）》2021年第2期127-131,共5页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：中国烟草总公司科技项目[110201901016(JY-03)];云南中烟工业有限责任公司资助项目(2016JC03);国家自然科学基金项目(21762050);云南省应用基础研究基金项目(2017FB014)。

摘　　要：在采集的尿液样品中依次加入内标和甲醇(控制样品介质中甲醇的体积分数约为50%),所得混合液经自主设计的样品离心分离和过滤装置处理,采用高效液相色谱-串联质谱法快速测定滤液中8-羟基脱氧鸟苷和8-羟基鸟苷的含量。以Synergi Polar-RP色谱柱为固定相,以不同体积比的10g·L^(-1)乙酸铵溶液和甲醇的混合液为流动相进行梯度洗脱,串联质谱分析中采用电喷雾离子源正离子模式和多反应监测模式。以15 N5-8-羟基脱氧鸟苷为内标。8-羟基脱氧鸟苷和8-羟基鸟苷的线性范围均为0.2~40μg·L^(-1),检出限(3S/N)依次为0.054,0.092μg·L^(-1),测定下限(10S/N)依次为0.18,0.32μg·L^(-1)。以中度吸烟者的尿液样品为基体进行加标回收试验,所得日内回收率、日间回收率分别为92.3%~96.3%,93.4%~96.3%,测定值的日内相对标准偏差(n=7)和日间相对标准偏差(n=7)分别为1.6%~3.0%,2.8%~3.4%。The collected urine sample was successively added with internal standard and methanol,and the volume fraction of methanol in the sample medium should be controlled to be about 50%.The above mixture was prepared by self-designed device for centrifugation and filtration of the sample.HPLC-MS/MS was applied to the rapid determination of 8-hydroxydeoxyguanosine and 8-hydroxyguanosine in the filtrate.Synergi Polar-RP chromatographic column was used as stationary phase,and the mixture of 10g·L^(-1)ammonium acetate solution and methanol mixed in different ratios was used as mobile phase in gradient elution.ESI+and multi-reactions monitor mode were adopted in MS/MS.15 N5-8-hydroxydeoxyguanosine was used as internal standard.Linearity ranges of 8-hydroxydeoxyguanosine and 8-hydroxyguanosine were found in the same range of 0.2-40μg·L^(-1),with detection limits(3S/N)of 0.054,0.092μg·L^(-1)and lower limits of determination(10S/N)of 0.18,0.32μg·L^(-1).On the base of urine sample of a moderate smoker,test for recovery was made by standard addition method;values of intra-day recovery and inter-day recovery were in the ranges of 92.3%-96.3%,93.4%-96.3%respectively,with intra-day RSDs(n=7)and inter-day RSDs(n=7)of determined values in the ranges of 1.6%-3.0%,2.8%-3.4%respectively.

关 键 词：高效液相色谱-串联质谱法 8-羟基脱氧鸟苷 8-羟基鸟苷 吸烟者 尿液 

分 类 号：O657.63[理学—分析化学]