食品科学

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顶空固相微萃取分析金黄色葡萄球菌挥发性代谢产物的条件优化

史 辉,唐俊妮,陈 娟*,王 琼   

  1. 西南民族大学生命科学与技术学院,四川 成都 610041
  • 出版日期:2015-06-25 发布日期:2015-06-12
  • 通讯作者: 陈 娟
  • 基金资助:

    国家自然科学基金面上项目(31371781);教育部“新世纪优秀人才支持计划”项目(NCET-11-0847);
    西南民族大学研究生创新项目(CX2014SP263)

Optimization of Headspace Solid Phase Micro-Extraction Conditions for GC-MS Analysis of  Volatile Metabolites from Staphylococcus aureus

SHI Hui, TANG Junni, CHEN Juan*, WANG Qiong   

  1. College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China
  • Online:2015-06-25 Published:2015-06-12
  • Contact: CHEN Juan

摘要:

采用65 μm聚二甲基硅氧烷/二乙基苯(polydimethylsiloxane/divinylbenzene,PDMS/DVB)、75 μm碳分子筛/聚二甲基硅氧烷(carboxen/polydimethylsiloxane,CAR/PDMS)萃取头和50/30 μm二乙基苯/碳分子筛/聚二甲基硅氧烷(divinylbenzene/carboxen/polydimethylsiloxane,DVB/CAR/PDMS)萃取头,20、30 min和40 min 3 种萃取时间,50、65 ℃和80 ℃ 3 种萃取温度,分别对金黄色葡萄球菌胰蛋白胨大豆肉汤(tryptone soy broth,TSB)培养物的挥发性代谢产物进行萃取,经气相色谱-质谱检测,以建立适宜于金黄色葡萄球菌挥发性代谢物的萃取分析方法。结果表明,50/30 μm DVB/CAR/PDMS萃取头能更好地满足各类化合物的萃取需要,获得更加全面的化合物信息,并且在萃取温度80 ℃条件下萃取30 min,得到的金黄色葡萄球菌挥发性代谢物的数量和响应强度较高。因此,萃取金黄色葡萄球菌挥发性代谢产物的最适条件为:50/30 μm DVB/CAR/PDMS萃取头、萃取温度80 ℃、萃取时间30 min。在该萃取条件下,检测出金黄色葡萄球菌在TSB中产生的主要挥发性物质为乙醇、1-癸醇、3-羟基-2-丁酮、丙酮、乙酸、苯甲醇、3-甲基-1-丁醇、吲哚、1-辛醇和正辛酸等。该萃取方法的建立可为细菌挥发性代谢产物的分析提供参考。

关键词: 金黄色葡萄球菌, 挥发性代谢产物, 萃取条件, 优化, 顶空固相微萃取-气相色谱-质谱联用

Abstract:

Volatile metabolites from Staphylococcus aureus cultured with tryptone soy broth (TSB) were extracted under
different conditions namely three types of fiber (65 μm PDMS/DVB fiber, 75 μm CAR/PDMS fiber and 50/30 μm DVB/
CAR/PDMS fiber), three extraction times (20, 30 and 40 min) and three extraction temperatures (50, 65 and 80 ℃),
respectively. The extracts were detected by gas chromatography-mass spectroscopy (GC-MS). The optimal extraction
conditions were determined. The results indicated that 50/30 μm DVB/CAR/PDMS fiber was suitable for the extraction of
various components and more comprehensive information was obtained using this fiber. Abundant amounts and response
intensities of bacterial volatile metabolites were extracted at 80 ℃ for 30 min. Accordingly the optimal extraction conditions
for volatile metabolites from S. aureus were obtained. Using the established extraction method, the main volatile compounds
produced by S. aureus cultured in TSB were detected to be alcohol, 1-decanol, 3-hydroxy-2-butanone, acetone, acetic acid,
benzyl alcohol, 3-methyl-1-butanol, indole, 1-octanol and octanoic acid. This extraction method can provide a valuable tool
for the analysis of bacterial volatile metabolites.

Key words: Staphylococcus aureus, volatile metabolites, extraction condition, optimization, headspace solid phase micro-extraction gas chromatography-mass spectroscopy (HS-SPME-GC-MS)

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