顶空气相色谱-质谱联用法分析粪便中挥发性脂肪酸  

Rapid Analysis of Volatile Fatty Acids in Feces by Headspace Gas Chromatography Tandem Mass Spectrometry

作　　者：江振作[1,2,3] 王跃飞[1,2] 陈荣荣[1,2] 朱彦[1,2] 张蕾[1,2] 刘双[1,2] 刘海利 

机构地区：[1]天津市现代中药重点实验室,天津中医药大学,天津300193 [2]中药新药研发中心,天津国际生物医药联合研究院,天津300457 [3]瀚盟生物技术(天津)有限公司,天津300457

出　　处：《分析化学》2014年第3期429-435,共7页Chinese Journal of Analytical Chemistry

基　　金：重大新药创制科技重大专项(No.2012ZX09304007);国家自然科学基金(No.81202877);天津市滨海新区专业性科技服务业项目(No.2012-XJR13006)资助~~

摘　　要：建立了一种快速分析粪便中挥发性脂肪酸（Volatile fatty acids,VFAs）的顶空气相色谱-质谱联用法（Headspace gas chromatography-mass spectrometry,HS-GC/MS）.粪便样品用6％ H3P04溶液按1∶2（m/V）混悬后密封于顶空进样瓶中,直接进行HS-GC/MS检测.顶空振荡室加热温度为80℃,振荡加热时间30 min,顶空进样针温度为80℃,不分流进样1 mL;采用DB-FFAP毛细管柱（30 m＆#215;0.25 mm＆#215;0.25 μm）,进样口温度为250℃,升温程序（初始温度50℃保持1 min,以10℃/min升至200℃）,载气（高纯氦）流速为1.0 mL/min;使用电子轰击（Electron impact,EI）离子源,电子能量为-70 eV,离子源温度为250℃,传输线温度为280℃,电子倍增器电压为0.95 kV,全扫描模式,扫描范围m/z 33 ～ 200.结果表明,本方法能够应用于人及大鼠粪便中挥发性脂肪酸的分析,通过NIST标准谱库检索,采用对照品比对以及质谱数据解析的方法,在人的粪便样品中检测到9种挥发性脂肪酸：乙酸、丙酸、异丁酸、丁酸、异戊酸、戊酸、异己酸、己酸、庚酸;在大鼠粪便中检测到8种挥发性脂肪酸：乙酸、丙酸、异丁酸、丁酸、异戊酸、戊酸、己酸、庚酸.通过峰面积归一化法,计算得到乙酸、丙酸、丁酸的相对百分含量约占挥发性脂肪酸总量的85％.本方法简单、灵敏,可用于人和大鼠粪便中挥发性脂肪酸的快速检测.A rapid headspace gas chromatography tandem mass spectrometric （HS-GC/MS） method was established for the analysis of volatile fatty acids （VFAs） in the feces. Feces were suspended by 6% phosphoric acid aqueous solution （ 1 ： 2 m/V） and sealed in the headspace bottle for HS-GC/MS analysis. The HS-GC/MS method was optimized as follows： agitator temperature （temp.） ： 80 ℃ , syringe temp. ： 80 ℃ , sample incubation time： 30 min, injection： 1 mL without split-flow. The chromatographic separation was performed on a DB-FFAP capillary column （30 m × 0.25 mm × 0.25 μm） with injection port temp. ： 250 ℃. The temperature program （initial temp. at 50 ℃ within first 1 min, and raised to 200 ℃ by 10 ℃/min） was employed by fixing the flow of carrier gas （ high purity helium） at 1.0 mL/min. The electron energy at -70 eV for electron impact （EI） ionization, ion source temp. ： 250 ℃, transfer line temp. ： 280 ℃, the voltage of electron multiplier at 0.95 kV. The spectra were recorded in the range of m/z 33-200 for full scan. The established HS-GC/MS method could be applied to analyze VGAs in the feces from human and rat appropriately. There are nine VFAs identified in the feces from human, and eight VFAs detected in the feces from rat by retrieving the NIST library, comparing with the standards and analyzing the MS data. Furthermore, the relative percentage contents of acetic acid, propionic acid and butyric acid accounted for roughly 85% of all VFAs by area normalization. The method is simple and sensitive, and it can be used to rapidly detect VFAs in the feces from human and rat.

关 键 词：顶空气相色谱-质谱联用法 粪便 肠道菌群 挥发性脂肪酸 

分 类 号：R446.1[医药卫生—诊断学]