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Supercritical fluid extraction of isoflavones from biological samples with ultra‐fast high‐performance liquid chromatography/mass spectrometry
Author(s) -
Klejdus Bořivoj,
Lojková Lea,
Lapčík Oldřich,
Koblovská Radka,
Moravcová Jitka,
Kubáň Vlastimil
Publication year - 2005
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500102
Subject(s) - daidzin , genistin , chromatography , glycitein , daidzein , chemistry , formononetin , mass spectrometry , isoflavones , extraction (chemistry) , high performance liquid chromatography , biochanin a , supercritical fluid extraction , genistein , sample preparation , medicine , biochemistry
An efficient method of modifier addition for supercritical fluid extraction (SFE) of polar isoflavones was developed and yielded extraordinarily high recoveries. To find the optimal extraction conditions, a temperature and pressure optimization and modifier impact study was performed in naturally contaminated and spiked samples. Ultra‐fast high‐performance liquid chromatography/mass spectrometry (HPLC/MS) was used for the determination of isoflavones on an Atlantis TM dC 18 high‐speed reversed phase chromatographic column (20×2.1 mm, 3 μm particle size). A newly elaborated supercritical fluid extraction (SFE) procedure allowed more accurate (< 5%) and precise (< 4–7%) determination of isoflavones in biological materials. The HPLC/MS method significantly reduced analysis time with simultaneous improvement of sensitivity and detection limits. The on‐column limits of detection LOD ( S/N = 3) for isoflavone glycosides (daidzin, genistin, glycitin, ononin, and sissotrin) were 1.3–3.6 fmol and 0.2–1.0 fmol for aglycones (daidzein, glycitein, genistein, formononetin, and biochanin A), respectively.