| Dipsaci radix is the dry roots of Dipsacus asper Wall.Ex Henry.It is named for its ability of healing broken bones and vessel.As a traditonal Chinese medicine,it is mainly used for treatment of pain in back,waist and leg,metrorrhagia,fetal movement and so on.Modern research results show that the main chemical composition in dipsaci radix including triterpenoid saponins,iridoid,volatile oil and alkaloids.Biological studies have revealed that this plant possesses antioxidant,anti-inflammatory,anticomplementary,cytotoxic effects and enhance immunity.The total saponins in dipsaci radix is the main active component making a significant role in promoting healing osteoporosis and fractures.This paper conducts the extraction and purification of total saponins in dipsaci radix and evalustes its quality in several components,in order to provide some reference for the follow-up study of dipsaci radix.This study examines the influence of several factors such as the extraction times,extraction solvent,extraction time and solvent dosage on the content of dipsaci radix total saponins.The extraction efficiency of total saponins was investigated with respecting to three variables including time,ethanol concentration and liquid-to-solid ratio.On the basis of a series of one-factor-at-a-time experiments,a polynomial regression model equation was fitted by the combined use of central composite experimental design and regression analysis.By analyzing the regression model using response surface analysis,the optimum extraction conditions of total saponins from dipsaci radix were identified as follows: 7 times of 55% alcohol,refluxing 3 times,3 hours every time,and the extraction efficiency of total saponins was up to 0.2011g/g and the asperosaponin ? was up to 0.0154g/g under the optimized conditions.Conclusion: Confirmatory experiment indicated the good prediction ability of the established model and provided the basis for further development and utilization of dipsaci radix.Compared the adsorption capacity of total saponins in dipsaci radix liquid of D101,AB-8 and DM-301 macroporous resins,D101 macroporous resin was chosed to purify and enrich total saponins in dipsaci radix.The adsorption and desorption rate of the total saponins in dipsaci radix are as the evaluation index.The final purification process is as follows: the sample concentration for crude drugs containing 0.25 g/mL of liquids,the velocity of sample with 2 bv/h,with 1.1 times column volume of sample liquid,with enough pure water rushed to colorless,7 times column volume and 70% ethanol elution with speed of 1 bv/h.By high performance liquid chromatography with UV assay(HPLC-UV),the semi-preparative HPLC,HPLC-MS and several other methods to prepare and analyze the main components from total saponins in dipsaci radix and to establish the content determination method.Extracts from total saponins in dipsaci radix by semi-preparative separation,manually collected eight compounds and four compounds are confirmed,respectively are loganin,glycosides A,?-heder and asperosaponin ?.The four compounds and the total saponin extracts from dipsaci radix in 10 batch from Hefeng and Yichang,hubei province determination method were constructed by HPLC.The scope of various compounds and the content of total saponins in extraction of dipsaci radix total saponins were respectively:loganin:0.3047%-0.9099%;glycosides A:0.5717%-4.1208%;?-heder:0.3183%-0.9423%;asperosaponin ?:3.4417%-9.8691%;the total saponins:50.5895%-67.4925%.HPLC chromatography was developed to establish the chromatographic fingerprint of total saponins in dipsaci radix.Eleven common peaks in chromatographic fingerprints were identified among 10 batches.All common peaks came out in 40 minutes and similarity was most more than 0.9.This provides a scientific basis for quality assessment of dipsaci radix total saponins extraction. |
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