血清脂溶性代谢物气相色谱质谱联用法检测与分析  

Detection and analysis of serum liposoluble metabolites by the combination of gas chromatography and mass spectrometry

作　　者：余好 陈江 罗小万 龙昌炜 徐纬[1] YU Hao;CHEN Jiang;LUO Xiaowan;LONG Changwei;XU Wei(Key Laboratory of Environmental Pollution Monitoring and Disease Control of Ministry of Education,School of Public Health,Guizhou Medical University,Guiyang 561113,Guizhou,China)

机构地区：[1]贵州医科大学公共卫生与健康学院,环境污染与疾病监控教育部重点实验室,贵州贵阳561113

出　　处：《贵州医科大学学报》2025年第1期31-38,共8页Journal of Guizhou Medical University

基　　金：2020年省级大学生创新创业训练计划项目资助(S202010660012);贵州省科技计划项目(黔科合LH字[2016]7378);贵州省区域内一流学科建设项目-公共卫生与预防医学(黔教科研发[2017]85)。

摘　　要：目的建立一种气相色谱质谱联用法(GC-MS)检测血清中脂溶性代谢物的分析方法。方法以SD大鼠为方法学研究对象,血清经乙酸乙酯漩涡萃取后,取萃取液100μL,加入1 g/L 1-氯萘10μL,氮吹至近干,加入15 g/L甲氧氨盐酸盐吡啶溶液50μL,70℃肟化1 h后氮吹至近干,加入含1%三甲基氯硅烷(TMCS)的N,O-双(三甲基硅基)三氟乙酰胺(BSTFA)100μL,低火力微波15 min,取溶液1μL注入GC-MS进行分析;以血清内源物质棕榈酸和内标物1-氯代萘作为考察对象,评价方法的检测限、线性关系、精密度和准确度;采用建立的GC-MS法对12只同批次健康SD大鼠(雌雄各半)血清进行检测,以确定各脂溶性代谢物与内标物峰面积的比值范围。结果本研究建立的GC-MS经方法学实验验证,棕榈酸和1-氯代萘的保留时间和峰面积的相对标准偏差(RSD)均<5%,1-氯代萘的加标回收率在95.8%~104.5%;对12只SD大鼠血清进行检测时,共检出24个共有色谱峰,鉴别得到13种脂溶性代谢物,并确定了各脂溶性代谢物与内标物峰面积的比值范围。结论本研究建立的GC-MS法对被检血清样品需求量小、检测成本低、精密度和准确度较好,能同时检测分析血清中多种脂溶性代谢物。Objective To establish a gas chromatography combined with mass spectrometry(GC-MS)technique to detect liposoluble metabolites in serum.Methods Sera of Sprague Dawley(SD)rats were selected as the methodological research object.The sera were extracted by vortex-assisted ethyl acetate to obtain the extraction solution.In 100μL of the extraction solution,10μL of 1 g/L 1-chloronaphthalene was added,blown to near dryness by nitrogen,then added with 50μL of 15 g/L methoxy ammonium hydrochloride pyridine solution,blown to near dryness by nitrogen after 1 hour of oximation reaction at 70℃,added with 100μL of N,O-Bis(trimethylsilyl)trifluoroacetamide(BSTFA)containing 1%trimethylchlorosilane(TMCS),microwaved at low power for 15 minutes.1μL of the solution was injected into GC-MS for analysis.Serum endogenous substance palmitic acid and internal standard 1-chloronaphthalene were selected as the investigation objects.The detection limit,linear relationship,precision and accuracy of the method was assessed.GC-MS was used to detect the sera of 12 healthy SD rats(half male and half female)from the same batch and determine the ratio ranges of the peak area of each lipid soluble metabolite to peak area of the internal standard in.Results GC-MS method established in this study was validated through methodological experiments,and the relative standard deviations(RSDs)of retention time and peak area for palmitic acid and 1-chloronaphthalene were both less than 5%.The standard recovery rate for 1-chloronaphthalene ranged from 95.8%to 104.5%.In the sera from 12 tested SD rats,a total of 24 common chromatographic peaks were detected,and 13 liposoluble metabolites were identified.The range of peak area ratios of each lipid soluble metabolite to the internal standard was determined.Conclusion The GC-MS established in this study has low demand for serum amount,low detection cost,good precision and accuracy.It can simultaneously detect and analyze multiple liposoluble metabolites in serum.

关 键 词：血清 代谢物 脂溶性 气相色谱质谱联用法 

分 类 号：R331[医药卫生—人体生理学]