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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.
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Objective:To develop a method for determining deoxynivalenol in four kinds of traditional Chinese herbal medicines. Methods:After being extracted by water, purified by an immunoaffinity column, deoxynivalenol was analyzed by LC-MS-MS. Results:The calibration curve was linear within the range of 2-50 ng·ml-1 for deoxynivalenol. The recovery was 68. 7%-88. 3%. Conclusion:The method is simple, sensitive and accurate in the determination of deoxynivalenol in Semen Ziziphi Spinosae, Semen Sojae Praepara-tum, Semen Coicis and Fructus Psoraleae.
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OBJECTIVE: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for simultaneous determination of daidzein, daidzin, genistein and genistin in Semen Sojae Praeparatum.
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Objective To analyze the fragments of 3 isoflavones and their isoflavone aglycones of Semen Sojae Praeparatum by electron spray ion trap mass spectrometry (ESI-MS), and to study the fragmentation pathway by major fragment ions. Methods Samples were fed into the instrument in the positive andnegative modes, and the fragments of the samples were yielded by multi-stage ion trap mass spectrometry (ESI-MS). The fragment ions of isoflavones and their isoflavone aglyconeswere analyzed. Results In the positive mode, ion peaks m/z 417, 255, 227, 199, 137, and 119 were detected for daidzin and daidzein; m/z 447, 285, 270, and 229 for glycitin and glycitein; and m/z 433, 271, 253, 243, 215, and 153 for genistin and genistein. In negative mode, ion peaksm/z 461, 415, 253, 225, 209, and 197 were detected for daidzin and daidzein; m/z 491, 445, 283, and 268 for glycitin and glycitein; and m/z 477, 269, 268, and 225 for genistin and genistein. Conclusion In the positive mode, daidzin and daidzein are fragmented by losing -Glu and -CO and Retro Diels-Alder (RDA) reaction; glycitin and glycitein are fragmented by losing -Glu, -CO, and -CH3; genistin and genistein are fragmented by losing -Glu, -CO, -H2O and RDA reaction. In negative mode, ions fragment [M+HCOO]- is produced by isoflavone glucosides, and daidzin and daidzein are fragmented by losing -Glu, -CO, -2CO and -CO2; glycitin and glycitein are fragmented by losing -Glu, -H, and -CH3; and genistin and genistein are fragmented by losing -Glu, -H, and-CO2.; glycitin amd glycitein are fragmented by losing -Glu-H, and -CH3; and genistin and genistein are fragmented by losing -Gli, -H, and -CO2.
ABSTRACT
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.