Simultaneous determination of six adenyl purines in human plasma by high-performance liquid chromatography with fluorescence derivatization
Simultaneous determination of six adenyl purines in human plasma by high-performance liquid chromatography with fluorescence derivatization
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DOI：
10.1016/s0378-4347(01)00265-1
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发表时间：
2001-08-25
期刊：
影响因子：
--
通讯作者：
Sato, H
中科院分区：
文献类型：
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作者：
Katayama, M;Matsuda, Y;Sato, H
建立了同时测定人血浆中6种腺苷嘌呤的高效液相色谱法。以腺嘌呤、腺苷、AMP、ADP、ATP和环AMP为荧光标记物，采用2-氯乙醛衍生化，对反应条件和分离条件进行了重新研究，以提高小体积样品分析的灵敏度。在Capcell Pack SG 120 A（TM）柱上分离每种衍生的嘌呤，其中移动的流动相由0.05 M柠檬酸-0.1 M磷酸氢二钾（pH 4.0）-甲醇（97+3）组成。荧光检测的检测限为100-1000 fmol/ml，比以前的报道高出约500倍。该方法已用于人血浆中腺苷嘌呤的测定。嘌呤水平如下：ATP（9.2-22.2 pmol/ml），ADP（5.5-22.2 pmol/ml），AMP（0.8-3.2 pmol/ml）。其他嘌呤、腺嘌呤、腺苷、cAMP均低于0.1 pmol/ml。(C)2001 Elsevier Science B. V.保留所有权利。
A sensitive method was developed for the simultaneous determination of six adenyl purines in human plasma by high-performance liquid chromatography. The adenyl purines (adenine, adenosine, AMP, ADP, ATP and cyclic AMP) were derivatized using 2-chloroacetaldehyde for fluorescence detection, and the reaction and separation conditions were reinvestigated to improve sensitivity for small volume sample analysis. Each derivatized purine was separated on a Capcell Pack SG120A(TM) column with mobile phase consisting of 0.05 M citric acid-0.1 M dipotassium hydrogen phosphate (pH 4.0)-methanol (97+3). The detection limits were 100-1000 fmol/ml by fluorescence detection, some 500 times better than previous reports. The proposed method was applied to determine adenyl purines in human plasma. The purine levels were as follows: ATP (9.2-22.2 pmol/ml), ADP (5.5-22.2 pmol/ml), AMP (0.8-3.2 pmol/ml). Other purines, adenine, adenosine, cAMP were lower than 0.1 pmol/ml. (C) 2001 Elsevier Science B.V. All rights reserved.