色谱

色谱

• 研究论文 • 上一篇    下一篇

气相色谱-质谱联用辅助鉴定细菌内毒素标准品菌种来源的纯度

岳丽娜1,2,李京华2,贺高红1,于万滢2,邵英光2,王俊德2

  

  1. 1.State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China; 2.Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian 116023, China
  • 收稿日期:2005-01-28 修回日期:2005-11-21 出版日期:2006-01-30 发布日期:1987-09-25
  • 通讯作者: 贺高红

Analysis of the Impurity of Bacterium Source of Standard Endotoxin by Gas Chromatography/Mass Spectrometry

YUE Lina1，, LI Jinghua2, HE Gaohong1, YU Wanying2, SHAO Yingguang2, WANG Junde2   

  1. 1.State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China; 2.Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian 116023, China
  • Received:2005-01-28 Revised:2005-11-21 Online:2006-01-30 Published:1987-09-25

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摘要：

为分析内毒素标准品菌种来源的纯度,建立了采用气相色谱-质谱联用法、以N，O-双三甲基硅三氟乙酰胺作为硅烷化试剂对待测物进行衍生、对内毒素标准品所含的3-羟基脂肪酸种类进行检测的方法。色谱柱为DB-5 (60 m×0.25 mm i.d.);载气为氦气，恒压，柱前压206 kPa；进样器温度250 ℃；柱箱初始温度为90 ℃，保持5 min，以5 ℃/min的升温速率升至280 ℃，保持5 min。进样量为1 μL，不分流进样。质谱离子源为电子轰击离子源（EI），离子源温度为250 ℃，接口温度为280 ℃。通过对内毒素标准品和大肠杆菌、绿脓杆菌、去离子水中3-羟基脂肪酸种类的比较,探讨了通过3-羟基脂肪酸种类来辅助判断内毒素标准品菌种来源的问题。结果显示，来源为大肠杆菌的9000 EU/支内毒素国家标准品中只含有3-羟基十四烷酸；20 EU/支的内毒素工作标准品中除含有3-羟基十四烷酸外，还含有3-羟基十二烷酸,说明其含有非大肠杆菌的细菌。

关键词: 3-羟基脂肪酸, 大肠杆菌 , 内毒素标准品, 气相色谱, 质谱

Abstract:

To analyse the impurity of bacterium source of standard endotoxin, 3-hydroxy fatty acid species in different endotoxin standards was determined by gas chromatography/mass spectrometry (GC/MS) using N,O-bis(trimethylsilyl)trifluoroacetamide as the silanizing reagent. GC/MS analysis was performed using a gas chromatograph equipped with a 60 m×0.25 mm i.d. DB-5 fused silica capillary column and an injector at 250 ℃. Helium was used as the carrier gas under a constant pressure of 206 kPa. The oven was programmed at a rate of 5 ℃/min from 90 ℃ (held for 5 min) to 280 ℃(held for 5 min). The sample size was 1 μL. The transfer line was kept at 280 ℃. The quadrupole mass spectrometer was operated in electron impact (EI) ionization mode, and the temperature of the source was kept at 250 ℃. The kind of 3-hydroxy fatty acids in 9000 EU/tube national standard endotoxin, 20 EU/tube working standard endotoxin, escherichia coli, pseudomonas aeruginosa and deionized water were determined to study the purity of bacterium source of the standard endotoxin. It was shown that 9000 EU/tube endotoxin standard and escherichia coli only contained 3-hydroxytetradecanoic acid. There was 3-hydroxydodecanoic acid in 20 EU/tube working standard endotoxin, which indicated the presence of impurity of bacterium source.

Key words: 3-hydroxy fatty acids, endotoxin standard, escherichia coli , mass spectrometry (MS), gas chromatography (GC)