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Objective To compare the contents variation of six flavonoids includingdaidzin，glycitin, genistin, daidzein, glycitein and genisteinin black beans, semifinished and finished Sojae Semen Praeparatum．Methods The contents of flavonoids were determined by HPLC，the condition were Diamonsil C18 column (4.6×250 mm, 5 μm) , column temperature 30 ℃, detection wavelength 260 nm, mobile phase 0.2% acetic acid water (A) - methanol (B), gradient elution, flow rate 1.0 ml/min．Results The linearity of this method to determine 6 isoflavones was good (r≥0.9993) within the determination range, and the recovery rate met the requirements. The RSD of precision, repeatability and stability experiment was less than 4%, 3%and 3%. The results of HPLC showed that the contents of six flavonoidsin Sojae Semen Praeparatum increased significantly compared with black beans. And, the contents of six flavonoids in finished Sojae Semen Praeparatum were slightly more than those in semifinished Sojae Semen Praeparatum. Conclusion The HPLC method established in this study could accurately determine the content of 6 isoflavones in Sojae Semen Praeparatum. The content of six isoflavones in black beans could be increased by the fermentation, and the combined isoflavones were transformed into free isoflavones during the fermentation process.
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OBJECTIVE: To study the chemical constituents and their proliferation activity on rat osteoblasts of Semen Sojae Praeparatum. METHODS: The compounds were isolated by chromatography on silica gel column, MCI and sephadex LH-20 column. Their structures were elucidated by spectral analysis. The proliferation test was performed by using rat osteoblasts with MTT assays in vitro. RESULTS: Twelve compounds were obtained and identified as daidzin(1), daidzein(2), genistein(3), genistin(4), glycitein(5), glycitin(6), apigenin(7), β-sitosterol(8), stigmasterol(9), campesterol(10), syringaldehyde(11), syringic acid(12), respectively. CONCLUSION: Compounds 5-11 are isolated from Semen Sojae Praeparatum for the first time. The five isoflavones show proliferation activity on rat osteoblasts, and aglycones proved to show better activity than glycosides.
RESUMEN
Objective:To search for a rapid and efficient method for the isolation of 3 kinds of isoflavone glucosides from the ethanol extract of Semen Sojae Praeparatum.Methods: The crude isoflavones were extracted by 75% aqueous ethanol after removing the oil by Soxhlet extraction.Then 1300 macroporous resin,water and different concentrations of aqueous ethanol(10%,20%,30%,40%,50%,70%,and 95%) were used to separate and elute the crude isoflavones.The fraction eluted by 40% aqueous ethanol was subjected to preparative HPLC analysis.The chromatographic conditions: A YWG C18(10.0 mm?200 mm,i.d.10 ?m) column was used;the mobile phase consisted of acetonitrile-water-acetic acid at volume ratio of 25751(V/V/V) and a flow rate of 3.0 ml/min;the injection volume was 750 ?l;and the detection wave length was set at 260 nm.Results: Daidzin,glycitin and genistin were rapidly separated with the purities over 99% as determined by external standard HPLC.Conclusion: This technique is simple and suitable for the isolation of daidzin,glycitin and genistin from Semen Sojae Praeparatum.