Ultrahigh-pressure liquid chromatography of isoflavones and phenolic acids on different stationary phases

• Published in: Journal of Chromatography A 2008-06, Vol.1195 (1), p.52-59
• Main Authors: Klejdus, B., Vacek, J., Lojková, L., Benešová, L., Kubáň, V.
• Format: Article
• Language: English
• Subjects: Acid hydrolysis; Agronomy. Soil science and plant productions; Analytical chemistry; Biological and medical sciences; Caffeic Acids - chemistry; Caffeic Acids - isolation & purification; Chemical constitution; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - methods; Coumaric Acids - chemistry; Coumaric Acids - isolation & purification; Cyanopropyl; Economic plant physiology; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Gallic Acid - analogs & derivatives; Gallic Acid - chemistry; Gallic Acid - isolation & purification; General pharmacology; Genistein - chemistry; Genistein - isolation & purification; Glycine max - chemistry; Hydroxybenzoates - chemistry; Hydroxybenzoates - isolation & purification; Isoflavones; Isoflavones - chemistry; Isoflavones - isolation & purification; Liquid chromatography; Medical sciences; Molecular Structure; Other chromatographic methods; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; Phenolic acids; Phenyl; Pisum sativum - chemistry; Plant Extracts - chemistry; Plant Extracts - isolation & purification; Plants; Propionates; Reversed phase; Soxhlet; Trifolium - chemistry; Vanillic Acid - chemistry; Vanillic Acid - isolation & purification
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2008.04.069

Complete separation of aglycones and glucosides of selected isoflavones (genistin, genistein, daidzin, daidzein, glycitin, glycitein, ononin, sissotrin, formononetin, and biochanin A) was possible in 1.5 min using an ultrahigh-pressure liquid chromatography (U-HPLC) on a different particular chemically modified stationary phases with a particle size under 2 μm. In addition, selected separation conditions for simultaneous determination of isoflavones together with a group of phenolic acids (gallic, protocatechuic, p-hydroxybenzoic, vanillic, caffeic, syringic, p-coumaric, ferulic, and sinapic acid) allowed separation of all 19 compounds in 1.9 min. Separations were conducted on a non-polar reversed phase (C 18) and also on more polar phases with cyanopropyl or phenyl groups using a gradient elution with a mobile phase consisting of 0.3% aqueous acetic acid and methanol. Chromatographic peaks were characterised using parameters such as resolution, symmetry, selectivity, etc. Individual substances were identified and quantified using UV–vis diode array detector at wavelength 270 nm. Limits of detection (3 S/N) were in the range 200–400 pg ml −1. Proposed U-HPLC technique was used for separation of isoflavones and phenolic acids in samples of plant materials ( Trifolium pratense, Glycine max, Pisum sativum and Ononis spinosa) after acid hydrolysis of the samples and modified Soxhlet extraction.