DICP OpenIR
学科主题分析化学
Determination of modified nucleosides in Escherichia coli by Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry
Xie YP(谢玉萍); Gao P(高鹏); Wang XY(王希越); Lian RW(练荣伟); Tian J(田晶); Xu GW(许国旺)
会议文集Proceeding of HPLC 2011
会议名称37th International Symposium on High Performance Liquid Phase Separations and Related Techniques
会议日期2011-10-8
2011
会议地点大连
页码503-0
出版者待补充
出版地待补充
合作性质墙报
部门归属1808
主办者中国化学会色谱专业委员会
英文摘要To date, there are about 100 modified nucleosides (MNs) have been identified in RNA species and the majority of them were found in tRNAs. Except their common properties as components of RNAs, MNs have been confirmed to be of multifunctional. Some researches had demonstrated MNs’ involvement in anti-stress responses in the yeast. Gene yfcC was recently found to modulate anaerobic survival of Escherichia. coli (E. coli) by us. In order to determine if disruption of yfcC could stimulate certain response manifesting by MNs, we developed a quick and simple method to quantify 20 nucleosides by using triple quadurupole liquid chromatography-tandem mass spectrometry (LC-MS/MS). Based on the consideration of the polar properties of the MNs, an Acquity UPLC HSS T3 column was adopted with the mobile phase of 10 mM ammonium acetate and acetonitrile at a flow-rate of 0.2 ml min-1. For the improvement of detection limit, multiple reaction monitoring mode as well as positive electrospray ionization was employed by using Triple Quadrupole mass spectrometry. The linearity of the detected MNs was excellent with R2 > 0.99 and the limits of detection were satisfied. The intra- and inter-day variation was ranged from 1.5% to 5.6% and 2.6% to 18.4%. Many of the included nuclesides could be readily quantified in E. coli and the most abundant of which was inosine. Additionally, distinct composition of the MNs could be distinguished between the wild type and genetic modified strains. This study indicated the effects of genetic manipulation on MNs in prokaryote also existed.; To date, there are about 100 modified nucleosides (MNs) have been identified in RNA species and the majority of them were found in tRNAs. Except their common properties as components of RNAs, MNs have been confirmed to be of multifunctional. Some researches had demonstrated MNs’ involvement in anti-stress responses in the yeast. Gene yfcC was recently found to modulate anaerobic survival of Escherichia. coli (E. coli) by us. In order to determine if disruption of yfcC could stimulate certain response manifesting by MNs, we developed a quick and simple method to quantify 20 nucleosides by using triple quadurupole liquid chromatography-tandem mass spectrometry (LC-MS/MS). Based on the consideration of the polar properties of the MNs, an Acquity UPLC HSS T3 column was adopted with the mobile phase of 10 mM ammonium acetate and acetonitrile at a flow-rate of 0.2 ml min-1. For the improvement of detection limit, multiple reaction monitoring mode as well as positive electrospray ionization was employed by using Triple Quadrupole mass spectrometry. The linearity of the detected MNs was excellent with R2 > 0.99 and the limits of detection were satisfied. The intra- and inter-day variation was ranged from 1.5% to 5.6% and 2.6% to 18.4%. Many of the included nuclesides could be readily quantified in E. coli and the most abundant of which was inosine. Additionally, distinct composition of the MNs could be distinguished between the wild type and genetic modified strains. This study indicated the effects of genetic manipulation on MNs in prokaryote also existed.
文献类型会议论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/116079
专题中国科学院大连化学物理研究所
通讯作者Xu GW(许国旺)
推荐引用方式
GB/T 7714
Xie YP,Gao P,Wang XY,et al. Determination of modified nucleosides in Escherichia coli by Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry[C]. 待补充:待补充,2011:503-0.
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