Preparative isolation and purification of two isoflavones from Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography

Peking University, Peping, Beijing, China
Journal of Chromatography A (Impact Factor: 4.17). 05/2003; 992(1-2):193-7. DOI: 10.1016/S0021-9673(03)00315-7
Source: PubMed


Two isoflavones, calycosin-7-O-beta-D-glycoside and formononetin-7-O-beta-D-glycoside, were separated from n-butanol extract of the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao by high-speed counter-current chromatography in two steps using two different solvent systems composed of ethyl acetate-ethanol-n-butanol-water (30:10:6:50, v/v) and ethyl acetate-ethanol-water (5:1:5, v/v). From 200 mg of crude extract, calycosin-7-O-beta-D-glycoside (12 mg) and formononetin-7-O-beta-D-glycoside (10 mg) were isolated at over 95% purity by HPLC analyses, and their structures were identified by MS, 1H NMR and 13C NMR.

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    • "Radix Astragali is widely used for its antiperspirant, antidiuretic, and antidiabetic properties and as a tonic drug [1] [2] [3]. It possesses various beneficial compounds, including astragalosides , isoflavonoids, isoflavones, isoflavan, and pterocarpan glycosides [4] [5] [6]. Due to the high market demand for Radix Astragali, a diverse group of adulterants with similar-morphological characteristics from genuses, such as Astragalus, Hedysarum, and Malva are often used in its stead [7]. "
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    Evidence-based Complementary and Alternative Medicine 08/2014; 2014:843923. DOI:10.1155/2014/843923 · 1.88 Impact Factor
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    • "It is believed by the consumers that the dried root reinforces ''Qi'' (vital energy) and is often used as an antiperspirant , an immunostimulant, a diuretic and as a supplementary medicine during cancer therapy (Zheng 2005; Wagner et al. 1997). The isoflavone compounds of roots are most often related to the bioactivity of the herb (Ma et al. 2003); calycosin and its 7-O-b-D-glu- coside are the major isoflavones in roots of A. membranaceus var. mongholicus (Wu et al. 2005). "
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    ABSTRACT: The changes in calycosin and calycosin-7-O-beta-D-glucoside content as well as the expression of genes involved in their biosynthesis were monitored in roots, stems and leaves of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao seedlings during 10 days of low temperature treatment. The concentrations of calycosin and its 7-O-beta-D-glucoside in the different tissues were analyzed using high-performance liquid chromatography. Higher glycoside contents were observed at 2 degrees C than that at 16 degrees C in all the tested tissues, however, the aglycone was scarcely detected in both leaves and stems at either 16 or 2 degrees C. cDNA fragments encoding four structural genes from the calycosin pathway, namely chalcone synthase, isoflavone synthase, isoflavone 3'-hydroxylase and UDP-glucose: calycosin-7-O-glucosyltransferase were isolated from A. membranaceus var. mongholicus seedlings by polymerase chain reaction (PCR) and sequenced. Real-time quantitative reverse transcript PCR demonstrated that in leaves and stems, five genes (including phenylalanine ammonia lyase), exhibited clear differences in their accumulation pattern in response to a low temperature stress, which was consistent with the increased content of calycosin-7-O-beta-D-glucoside. In the roots, transcription of the five genes was down-regulated at 2 degrees C, but the contents of calycosin and its glucosides were higher than that at 16 degrees C. These findings indicate that low temperature stress could induce accumulation of calycosin and its glucosides in different tissues of the seedlings of A. membranaceus var. mongholicus but the mechanisms regulating the accumulation were different.
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