Zgórka Grazyna
Department of Pharmacognosy with Medicinal Plant Unit, Medical University of Lublin, 1 Chodzki Street, 20-093 Lublin, Poland.
J Sep Sci. 2009 Apr;32(7):965-72. doi: 10.1002/jssc.200800456.
Detailed chemotaxonomic studies were undertaken to establish the qualitative profile and real amounts of the pharmacologically active isoflavone aglycones genistein, daidzein, formononetin, and biochanin A in aerial parts of thirteen Trifolium L. (clover) species, native to Poland. A newly elaborated micropreparative technique--SPE--on BakerBond octadecyl, cyclohexyl, and phenyl cartridges was used in combination with ultrasound-assisted extraction for isolation of isoflavone aglycones from hydrolyzed samples. The effectiveness of all three SPE sorbents in the purification of plant extracts was compared and very high recoveries (>96%) were documented for four isoflavones. Classical photodiode-array and very sensitive fluorescence detection, coupled with reversed-phase high-performance liquid chromatography (RP-HPLC), were employed to obtain the most reliable qualitative and quantitative results. Chemotaxonomic differences combined with flower color variability were demonstrated within thirteen clover species. Concentration levels of particular isoflavones in ten Trifolium species possessing flowers with white, pink, or purple-red corolla ranged from approximately 3 to approximately 3300 microg/g dry weight, while in three yellow flowering clovers (T. aureum, T. dubium, and T. campestre) isoflavone compounds have not been detected at all. RSD values, determined for intra- and inter-day precision of the quantitative results, were not higher than 6.2% and 7.1%, respectively.
开展了详细的化学分类学研究，以确定原产于波兰的13种三叶草属（Trifolium L.）植物地上部分中具有药理活性的异黄酮苷元染料木黄酮、大豆苷元、芒柄花黄素和鹰嘴豆芽素A的定性概况及实际含量。一种新开发的微量制备技术——固相萃取（SPE），采用BakerBond十八烷基、环己基和苯基柱，结合超声辅助萃取，从水解样品中分离异黄酮苷元。比较了所有三种SPE吸附剂在纯化植物提取物中的效果，四种异黄酮的回收率都很高（>96%）。采用经典的光电二极管阵列和非常灵敏的荧光检测，结合反相高效液相色谱（RP-HPLC），以获得最可靠的定性和定量结果。在13种三叶草物种中证明了化学分类学差异与花色变异性的结合。10种具有白色、粉色或紫红色花冠花朵的三叶草物种中，特定异黄酮的浓度水平范围为约3至约3300微克/克干重，而在三种开黄花的三叶草（金黄三叶草、小巢菜和野火球）中，根本未检测到异黄酮化合物。定量结果日内和日间精密度的相对标准偏差（RSD）值分别不高于6.2%和7.1%。
