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	Development of an ion-pair HPLC method for investigation of energy charge changes in cerebral ischemia of mice and hypoxia of Neuro-2a cell line
	Chen, Yunyun ; Xing, Dongming ; Wang, Wei ; Ding, Yi ; Du, Lijun
	2010-05-11 ; 2010-05-11
关键词	IP-HPLC energy charge adenine nucleotides cerebral ischemia Neuro-2a cell line PERFORMANCE LIQUID-CHROMATOGRAPHY RAT-BRAIN INVIVO ADENINE-NUCLEOTIDES REVERSED-PHASE ATP NUCLEOSIDES DEOXYNUCLEOTIDES METABOLITES EXTRACTION SYSTEM Biochemical Research Methods Biochemistry & Molecular Biology Chemistry, Analytical Pharmacology & Pharmacy
中文摘要	The determination of adenine nucleoticles and energy charge (EC) has great importance in the characterization of cerebral ischemic injury and post-ischemic recovery. An IP-HPLC method was developed for the quantification of AMP, ADP, ATP and EC in cerebral ischemia and hypoxia of the Neuro-2a cell line. The chromatographic conditions were: a Zorbax SB-C-18 reversed-phase column: mobile phase 100 mm KH2PO4, 1 mm tetrabutyl ammonium hydroxide, and 2.5% acetonitrile. brought to pH 7.0 with potassium hydroxide (4 m), filtered through a 0.45 mu m Millipore filter and degassed prior to use. The flow-rate was 1.0 mL/min. The injection volume was 20 mu L. Detection was performed at a wavelength of 254 nm under a constant temperature (27 VC). The method was validated by means of linearity, using calibration curves constructed with five concentration levels of each compound. The limit of detection was also determined. The system precision was calculated as the coefficient of variation for five injections for each compound tested. Cerebral tissue was homogenized (4 degrees C) in 1 mL of an ice-cold 6% trichloroacetic acid that contained ATPase inhibitor and obtained good recovery (> 90%). The results show that the described method for the determination of adenine nucleotides by HPLC has good linearity, limit of detection, precision and specificity, and is simple and rapid to perform. Copyright (c) 2007 John Wiley & Sons. Ltd.
语种	英语 ; 英语
出版者	JOHN WILEY & SONS LTD ; CHICHESTER ; THE ATRIUM, SOUTHERN GATE, CHICHESTER PO19 8SQ, W SUSSEX, ENGLAND
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/27034]
专题	清华大学
推荐引用方式 GB/T 7714	Chen, Yunyun,Xing, Dongming,Wang, Wei,et al. Development of an ion-pair HPLC method for investigation of energy charge changes in cerebral ischemia of mice and hypoxia of Neuro-2a cell line[J],2010, 2010.
APA	Chen, Yunyun,Xing, Dongming,Wang, Wei,Ding, Yi,&Du, Lijun.(2010).Development of an ion-pair HPLC method for investigation of energy charge changes in cerebral ischemia of mice and hypoxia of Neuro-2a cell line..
MLA	Chen, Yunyun,et al."Development of an ion-pair HPLC method for investigation of energy charge changes in cerebral ischemia of mice and hypoxia of Neuro-2a cell line".(2010).

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