Immunoaffinity chromatography of abscisic acid combined with electrospray liquid chromatography-mass spectrometry

被引:41
作者
Hradecka, Veronika
Novak, Ondrej
Havlicek, Libor
Strnad, Miroslav
机构
[1] Palacky Univ, Lab Growth Regulators, CZ-78371 Olomouc, Czech Republic
[2] ASCR, Inst Expt Bot, CZ-78371 Olomouc, Czech Republic
[3] ASCR, Inst Expt Biol, Isotope Lab, Prague 14220, Czech Republic
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2007年 / 847卷 / 02期
关键词
abscisic acid; immunoaffinity chromatography; liquid chromatography-mass spectrometry;
D O I
10.1016/j.jchromb.2006.09.034
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Polyclonal antibodies with high specificity for C1-immobilised (+)-cis,trans-abscisic acid (ABA) were raised, characterised by enzyme-linked immunosorbent assay (ELISA) and used for preparation of an inummoaffinity chromatography (IAQ gel. The detection limit of the ELISA was approximately 4.6 x 10(-10) mol/L. Sensitive electrospray liquid chromatography-mass; spectrometry (LC-ESI-MS) methods, were also developed with detection limits below 0.1 x 10(-12) mol. The IAC allowed quick, single-step processing of samples prior to the analyses. The LC-ESI-MS and LC-ELISA techniques were used for comparative estimation of endogenous ABA levels in immumoaffinity purified extracts of normal and water-stressed Nicotiana tabacum L. leaves. The analytical approaches were validated using deuterium- and tritium-labelled internal standards, respectively. The LAC method was found to be highly effective, sensitive and convenient for isolating the target analyte from plant material. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:162 / 173
页数:12
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