Research On Quality Standard Of Melandrium Rohrb

ObJctive To study the pharmacognositic identification of Melandrium firmum Rohrb.,a Chinese Chaoxianzu folk medicine,to establish the TLC and HPLC finger-print,and to detect the content of four flavonoid glycosides.Methods 1 Pharmacognositic identification of M.firmum was carried out by origin,characteristics and microscopic identification methods.2 TLC and HPLC finger-print were established.HPLC separation of analysis was achieved on a Waters SunFireTM C18 column(5μm,4.6mm×250mm,Ireland).The detection wavelength was set at 253nm and the column temperature was set as 40℃.The mobile phase consisted of 0.4%(v/v)glacial acetic acid in distilled water(A)and acetonitrile(B),gradient elution.The flow rate of 1.0 mL/min.The injection volume was 10 μL.3 Separated and purified the chemical constituents from the n-butanol extract of M.firmum,three flavonoid glycoside were obtained and were used as standards.4 In this paper,the content of the extract of M.firmum samples was determined by HPLC,with apigenin-8-C-arabinofuranosides,isovitexin,chrysoeriol-8-C-arabinofuranoside and isoscoparin.Results 1 The microscopic characteristics of the stem,leaf and powder of M.firmum herba were described in detail.The powder showed non-glandular hairs,glandular hairs,cork cells,calcium oxalate crystals and other structures.The phloem of the stem was slightly degenerated,the xylem was mostly arranged in radial direction,and the medulla was broad,with about 7 layers of cortical cells loosely arranged.The leaf cross-section was shown in 1 column of palisade tissue,and the vascular bundle was the external toughness type.2 The TLC fingerprint of the M.firmum was preliminarily established,two common spots were better reproducible at 365nm and 254nm.3 The datum of 10 batches of M.firmum samples obtained from different places was analyzed by Chinese medicine chromatographic fingerprint similarity evaluation system 2004A version,and calibrated with 21 characteristic peaks,established a common pattern of HPLC fingerprint.4 Three flavonoid glycosides were extracted from the n-butanol extract of M.firmum,through positive silica gel column chromatography,MPLC,and other separation methods.Three compounds were identified as apigenin-8-c-arabinofuranosides,chrysoeriol-8-c-arabinofuranoside and isoscoparin.Three compounds were isolated from this plant for the first time and were used as maker compounds.5 Determination of HPLC-UV content:all of the 10 batches of M.firmum herbs contained the four compounds,but the contents were differences.The results indicated that the contents of apigenin-8-C-arabinofuranosides(0.12-0.98 mg/g),isovitexin(0.17-0.49 mg/g),chrysoeriol-8-C-arabinofuranoside(0.24-3.08 mg/g),isoscoparin(0.21-7.78 mg/g)in 10 herbal samples.Conclusion By studying the identification of origin,character,microscopic characteristics,TLC and HPLC fingerprints and content determination of M.firmum,the quality standards of this Korean Chinese folk medicine were established,to filled the gap of quality standards of Melandrium firmum Rohrb.