Comparison of a Commercial ELISA Assay for Indole-3-Acetic Acid at Several Stages of Purification and Analysis by Gas Chromatography-Selected Ion Monitoring-Mass Spectrometry Using a 13C6-Labeled Internal Standard | Zendy

Jerry D. Cohen | Zendy; Michael G. Bausher | Zendy; Krystyna Białek | Zendy; J. George Buta | Zendy; Gregory F. W. Gocal | Zendy; Loeke Janzen | Zendy; Richard P. Pharis | Zendy; Arden N. Reed | Zendy; Janet P. Slovin | Zendy

Open Access

Comparison of a Commercial ELISA Assay for Indole-3-Acetic Acid at Several Stages of Purification and Analysis by Gas Chromatography-Selected Ion Monitoring-Mass Spectrometry Using a ¹³C₆-Labeled Internal Standard

Author(s) -

Jerry D. Cohen,

Michael G. Bausher,

Krystyna Białek,

J. George Buta,

Gregory F. W. Gocal,

Loeke Janzen,

Richard P. Pharis,

Arden N. Reed,

Janet P. Slovin

Publication year - 1987

Publication title -

plant physiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 3.554

H-Index - 312

eISSN - 1532-2548

pISSN - 0032-0889

DOI - 10.1104/pp.84.4.982

Subject(s) - chromatography , mass spectrometry , chemistry , quantitative analysis (chemistry) , gas chromatography , acetic acid , gas chromatography–mass spectrometry , indole 3 acetic acid , high performance liquid chromatography , benzene , biochemistry , organic chemistry , gene , auxin

Quantitative analysis of indole-3-acetic acid (IAA) using selected ion monitoring gas chromatography-mass spectrometry (GC-MS) with (13)C(6)[benzene ring]-IAA as the internal standard was used to compare the quantitative accuracy of commercial enzyme-linked immunoabsorbent assay (ELISA) kits. Plant materials differed in the amount of purification required prior to use of ELISA for reliable estimates to be made. Purification similar to that obtained by at least one high performance liquid chromatographic (HPLC) step was generally necessary prior to ELISA analysis of plant materials. Additional levels of purification appeared to be required for some plant materials prior to HPLC in order to obtain an accurate estimate by ELISA techniques. In no case was it possible to obtain reasonable estimates of IAA from crude extracts or even from acidic fractions of extracts of plant tissues. GC-MS techniques provide a rapid and simple method for checking the validity of ELISA techniques. Quantitative GC-MS, or a similar technique that provides an independent quantitative validation, should, whenever possible, be applied to each new plant material under study if use of the ELISA is planned.

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