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Analysis of IAA and its metabolites by immunoaffinity extraction and HPLC-MS/MS

IAA and its metabolites occur in a very complex matrix of plant samples at diverse concentrations, ranging from tens of nmol to hundreds of fmol per g of fresh weight. To isolate and quantify the metabolites, we developed an analytical protocol combining a highly-specific anti-IAA immunoaffinity extraction with a sensitive and selective LC-MS/MS analysis. By using internal standards for each of the analyzed compounds, we can study IAA and a wide range of its metabolites including amino acid conjugates. The protocol enables to quantify the metabolites in amounts of fresh material as low as 20 mg.

Immunoaffinity extraction; High-performance liquide chromatography (HPLC); Tandem mass spectrometry (MS/MS); Indole-3-acetic acid (IAA); Auxin conjugates

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