Determination of 18 phenolic acids in tobacco and rhizosphere soil by ultra high performance liquid chromatography combined with triple quadrupole mass spectrometry

An ultra high performance liquid chromatography with triple quadrupole mass spectrometry method for the determination of free and bound phenolic acids in tobacco plant and soil was developed. A simple solid-phase extraction, which used Polar Enhanced Polymer column as stationary phase and methanol as mobile phase, was used for the clean-up of bound phenolic acids, and a liquid-phase extraction using chloroform as solvent was used to purify free phenolic acids. With our method, 18 phenolic acids in rhizosphere soil of continuous cropping flue-cured cultivar k326 were separated and determined within 6 min with recoveries of 82-107% and relative standard deviations (n = 5) of 1.1-4.8%. Results showed that free phenolic acids accounted for 0-9, 92-100, and 69-100% of total phenolic acids in rhizosphere soil, cultivar k326 roots and leaves, respectively. Results also revealed that p-hydroxybenzoic acid, p-coumaric acid, vanillic acid, ferulic acid, and syringic acid were the predominant phenolic acids in rhizosphere soil of cultivar k326, and continuous cropping of cultivar k326 in the same farmland could lead to the accumulation of these phenolic acids in soil except syringic acid. The determination of phenolic acids provided detailed information for evaluating their source and characteristics in continuous cropping tobacco plant and soil.