Development of a Rapid LC-DAD/FLD Method for the Simultaneous Determination of Auxins and Abscisic Acid in Plant Extracts

Abstract

Plant hormones play a crucial role in controlling plant growth and development. These groups of naturally occurring substances trigger physiological processes at very low concentrations, which mandate sensitive techniques for their quantitation. This paper describes a method to quantify endogenous (±)-2-cis-4-trans-abscisic acid, indole-3-acetic acid, indole-3-propionic acid, and indole-3-butyric acid. The method combines high-performance liquid chromatography (HPLC) with diode array and fluorescence detection in a single run. Hybrid tea rose 'Monferrato' matrices (leaves, petals, roots, seeds, androecium, gynoecium, and pollen) were used as references. Rose samples were separated and suspended in extracting methanol, after which (±)-2-cis-4-trans-abscisic acid and auxins were extracted by solvent extraction. Sample solutions were added first to cation solid phase extraction (SPE) cartridges and the eluates to anion SPE cartridges. The acidic hormones were bound to the last column and eluted with 5% phosphoric acid in methanol. Experimental results showed that this approach can be successfully applied to real samples and that sample preparation and total time for routine analysis can be greatly reduced.

Dynamics of abscisic acid and indole-3-acetic acid during the early-middle stage of seed development in Rosa hybrida

“…During hybridisation and hip development, monthly mean greenhouse temperatures were measured as 19.8°C Analysis of plant growth regulators by HPLC ABA and IAA were assessed at 28, 35, 42, and 49 DAP in the seed coat and embryo of R. hybrida by HPLC based on SPE purification, according to Bosco et al (2013). Analytical reagent grade chemicals were used, unless otherwise indicated.…”

“…Then, 0.3 g of each homogenized sample was suspended with 2 mL of 80 % aqueous methanol containing 10-20 mg L -1 of butylated hydroxytoluene for 16 h at 4°C in darkness under magnetic stirring. Extraction was performed as described by Bosco et al (2013) using SPE DSC-MCAX and SPE LC-NH2 cartridges. A total of 20 lL of purified samples were injected into HPLC.…”

“…In this work, we used an analytical method that allowed simultaneous detection of both ABA and IAA amounts. To clearly illustrate the dynamics of ABA and IAA in rose seed coat and embryo during seed development, plant growth regulators were firstly separated by their hydrophobic and acidic properties using anionic SPE, and then separated with HPLC (Bosco et al 2013).…”

Localized surface plasmon resonance-based abscisic acid biosensor using aptamer-functionalized gold nanoparticles

“…So the quantitative determination of ABA is very essential for the physiological process of plants. Up to now, a number of analytical techniques have been developed for the detection of ABA, such as capillary electrophoresis [ 3 ], electrochemical immunosensors [ 4 – 6 ], high performance liquid chromatography [ 7 ] and liquid chromatography/mass spectrometry methods [ 8 – 11 ]. However, these methods have their inherent disadvantages, such as sophisticated instrument dependence and time-consuming procedures.…”

Simultaneous column chromatographic extraction and purification of abscisic acid in peanut plants for direct HPLC analysis

“…Although a variety of mixed organic solutions and pH buffer solutions were used for ABA extraction in different plant materials, methanol aqueous solution is the mostly used solvent to extract ABA from plants [8][9][10]. Traditionally, ABA was extracted by two or three times of the solvent macerating extraction followed by one or more steps of purification such as liquid-liquid fractionation or solid phase extraction, before used for content determination [7,[11][12][13][14].…”