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Plant Physiology 68:96-98 (1981)
© 1981 American Society of Plant Biologists

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Articles

Validation of a Radioimmunoassay for Indole-3-Acetic Acid Using Gas Chromatography-Selected Ion Monitoring-Mass Spectrometry 1

William L. Pengelly, Robert S. Bandurski2 and Aga Schulze

Department of Botany and Plant Pathology, Michigan State University, East Lansing, Michigan 48824

A radioimmunoassay for indole-3-acetic acid (IAA) has been validated by comparison with a physico-chemical assay utilizing gas chromatography-selected ion monitoring-mass spectrometry and 4,5,6,7-tetradeutero-indole-3-acetic acid as an internal standard. The radioimmunoassay provided a reliable estimate of the free IAA content of etiolated corn shoots. However, base hydrolysis of extracts for determination of ester IAA released substances which interfered with the radioimmunoassay. Interference was detected by internal controls and by lack of agreement with the mass spectral method. Interfering compounds could be removed from extracts by chromatography on diethylaminoethyl- and hydroxypropylated (lipophilic) Sephadex G-25. Following such purification the radioimmunoassay estimate of the total (free + ester) IAA content of etiolated corn shoots agreed with the mass spectral method within 2% on the average.

2 To whom reprint requests should be directed.

1 This work was supported by the Metabolic Biology Section of the National Science Foundation, PCM 79-04637, and is Journal Article 9693 from the Michigan Agricultural Experiment Station.






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Copyright © 1981 by the American Society of Plant Biologists