Determination of Salicinoids by Micro-high-performance Liquid Chromatography and Photodiode Array Detection
Article first published online: 26 NOV 2013
Copyright © 2013 John Wiley & Sons, Ltd.
Volume 25, Issue 3, pages 185–191, May/June 2014
How to Cite
Rubert-Nason, K. F., Hedman, C. J., Holeski, L. M. and Lindroth, R. L. (2014), Determination of Salicinoids by Micro-high-performance Liquid Chromatography and Photodiode Array Detection. Phytochem. Anal., 25: 185–191. doi: 10.1002/pca.2485
- Issue published online: 14 APR 2014
- Article first published online: 26 NOV 2013
- Manuscript Accepted: 6 OCT 2013
- Manuscript Revised: 1 OCT 2013
- Manuscript Received: 10 APR 2013
- capillary HPLC;
- phenolic glycoside;
Plant-derived salicinoids (conjugates of glucose and salicylate phenolic moieties) are potent modulators of plant–herbivore interactions. We demonstrate the use of micro-high-performance liquid chromatography (μHPLC) with photodiode-array detection (DAD) for quantification of four salicinoids (salicin, salicortin, hydroxycyclohexen-on-oyl salicortin and tremulacin) in methanolic extracts of Populus.
To develop and implement a solvent-conserving μHPLC method to quantify salicinoids in methanolic extracts of Populus tissue.
Salicinoids were extracted from Populus tissue into methanol, filtered, and introduced to μHPLC. Extracted analytes were separated on a Zorbax SB C18-column with a binary gradient of methanol and water (with 2% tetrahydrofuran; 20 μL/min), and quantified by DAD (274 nm). We confirmed measurement reliability through standard addition, comparison with an accepted method, and assessment of chromatographic peak purity by ultraviolet absorbance spectra.
Method detection and quantification limits for the salicinoids as a percentage of dry leaf weight were as follows: salicin (0.1%, 0.2%), salicortin (0.001%, 0.02%), hydroxycyclohexen-on-oyl salicortin (0.02%, 0.06%) and tremulacin (0.0006%, 0.002%). Calibrations by external standardisation were linear over 1.5 orders of magnitude with acceptable accuracy and reproducibility.
Micro-HPLC can serve as a solvent-conserving alternative to conventional HPLC for quantification of salicinoids in Populus tissue. Copyright © 2013 John Wiley & Sons, Ltd.