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目的:建立六神曲的指纹图谱质控方法,比较鲜干品各组样品发酵前后槲皮苷、槲皮素、木犀草素等成分的变化状况。方法:经拆方研究,按前期优选的工艺制备六神曲基本组及鲜干品各组,分别为基本组、鲜品煎汁组、干品1/3量煎汁组及干品全量煎汁组,并于发酵前后取样检测。采用LC-MS检测,Ulitimate XB-C18色谱柱(4.6 mm×250 mm,5μm),流动相甲醇-0.2%甲酸水溶液梯度洗脱,检测波长254 nm,电喷雾离子源(ESI),正离子模式检测。结果:确定鲜干品组方六神曲指纹图谱共有峰15个,其中10,13,15号峰分别为槲皮苷、木犀草素、槲皮素。基本组仅含木犀草素,发酵前后质量分数分别为18.5,21.3μg·g~(-1),明显低于其他3组。在发酵前,鲜品煎汁组样品的3种成分含量均显著高于其他3组,与发酵前相比,发酵后木犀草素、槲皮素质量分数分别增长54.84%,4.53%;槲皮苷质量分数降低了0.76%。干品全量煎汁组发酵后3种成分含量较发酵前显著增加,其中槲皮苷、木犀草素、槲皮素质量分数分别增加了59.9%,161.7%,75.5%。结论:该质控检测方法精密、准确、重复性好,适用于六神曲的量化质量评价。发酵法可促进六神曲部分药效成分的溶出度增加及其生物转化反应的发生,初步揭示了六神曲发酵前后药效物质变化状况及鲜品入药优于干品的药效物质基础。
OBJECTIVE: To establish a quality control method for fingerprint of Liushen Qu and to compare the changes of quercitrin, quercetin, luteolin and other components before and after fermentation of fresh and dried samples. Methods: According to the study of demolition, according to the preferential process, the basic group and the fresh dried products of each group were prepared, which were the basic group, the fresh juice decoction group, the dry decoction 1/3 decoction group and the dry decoction whole juice Group, and sampling before and after fermentation test. LC-MS, Ulitimate XB-C18 column (4.6 mm × 250 mm, 5 μm), gradient elution with mobile phase methanol-0.2% formic acid aqueous solution, detection wavelength 254 nm, electrospray ionization source (ESI), positive ion mode Detection. Results: The fingerprints of Fangliu Qu of Xiangan group were identified as 15, of which 10, 10 and 15 were the peaks of quercitrin, luteolin and quercetin, respectively. The basic group contained only luteolin, before and after fermentation mass fraction were 18.5,21.3μg · g ~ (-1), significantly lower than the other three groups. Before fermenting, the content of three components of fresh juice decoction group was significantly higher than that of the other three groups, and the content of luteolin and quercetin increased by 54.84% and 4.53% Glycosides decreased by 0.76%. After fermentation, the content of three components increased significantly after fermentation. The contents of quercitrin, luteolin and quercetin increased by 59.9%, 161.7% and 75.5%, respectively. Conclusion: The quality control method is accurate, accurate and reproducible. It is suitable for quantitative quality evaluation of Liushen Qu. Fermentation can promote the dissolution of some active ingredients of Liushen Qu and its bioconversion reaction, and reveal the changes of the efficacy substances before and after the Liushen Qu and the medicinal material basis of the fresh drugs into the dried products.