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Purification and determination of 3-indole-butyric acid and alpha-napthaleneacetic acid from plant tissues using solid phase extraction and high performance liquid chromatography

Author(s): C.Theerakarunwong, S.Phanichphant

The method for separation and purification of auxin compounds based on reversed phase solid phase extraction and high performance liquid chromatography (RP-SPE-HPLC)was investigated. The auxinswhichwere 3-indolebutyric acid (IBA) and alpha-napthaleneacetic acid (alpha-NAA) were separated using a reversed phaseC18column of150 x 4.6mmand 5mparticle. IBA and alpha-NAA can be completely separated at the flowrate of 0.7 mLmin-1 withmethanol/1%formic acid inwater (65:35 v/v) used as amobile phasewith the analysis time of 9.0 min. Solid phase extraction (SPE) was performed to pre-concentrate auxin compounds. Percent recoveries of the extractionmethod were found to be 89.0%. Plant tissueswere extracted using solvent extraction followed byC18SPE and then analyzed byHPLC. By spiking various concentrations ofmixed standard into plant tissues, percent recoveries of both analyte were obtained in the range of 81.5-105. The detected IBA was found to be in the range of 0.65-2.84mgmL-1 and 1.04-3.15mgmL-1 for alpha -NAA.Due to its overall analytical performance, these SPE and HPLC methods are presently being proved to be simple and suitable for routine purposes such as those developed for plant extracts studies.

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