An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.

OBJECTIVE:To establish a method for the content determination of saccharide in polysaccharides containing galact-uronic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS:The determination condi-tion of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%,the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide,and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pi-losula polysaccharides CPP1b. RESULTS:The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33;in vali-dation test,the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS:The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.

OBJECTIVETo develop a HPLC method for determination of matrine, oxymatrine; ferulic acid, L-shikonin and beta, beta-dimethylacrylshikonin in compound preparatioti of Dangguishuan.

METHODThe chromatogtaphic separation was performed on a Kromasil ODS C18 column (4.6 mm x 250 mm, 5 microm) maintaining at 30 degrees C during the whole process. The mobile phase consisted of methanol and 0.1% triethylamine aqueous solution (adjusted with phosphoric acid, pH 3) at a flow rate of 1.0 mL x min(-1). The detection wavelength was set at 220 nm for matririne and oxymatrine, 316 nm for ferulic acid, 516 nm for L-shikonin and beta, beta-dimethylacrylshikonin, respectively.

RESULTAll the compounds showed good linearity (r > 0.9996) in the range of the test concentrations, and the average recoveries of the method is betwuen 96.92% and 102.22%, RSD < 3.1%.

CONCLUSIONThe method is proved to be credible, sensitive, accurate and repeatable. It can be applied to determine of matrine, oxymatrine, ferulic acid, L-shikonin and beta, beta-dimethylacrylshikonin in compound preparation of Dangguishuan simultaneously, and provide a basal method of quality control to this preparation and other relative preparations.

Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Drugs, Chinese Herbal ; analysis ; Naphthoquinones ; analysis ; Quinolizines ; analysis

OBJECTIVETo determine such molecular characteristic parameters as absolute molecular weight, molecular weight distribution, root-mean-square turning radius (Rg) and polydispersity index (Mw/Mn) of four components contained in Hedysari Radix polysaccharide 3 (HPS-3) and map weight-average molecular weight (Mw) with root-mean-square turning radius (Rg), in order to calculate conformations of the four components at solution state.

METHODThe gel permeation chromatography-multi angle laser light scatting (GPC-MALLS) was adopted, with 0.1 mol x L(-1) NaNO3 contained 0.02% NaN3 as the mobilephase, Ultrahydrogel 1000 connected in series with Ultrahydrogel500.

RESULTAmong the four components of HPS-3, HPS-3-C showed the highest weight average molecular weight of 1.986 x 10(5) g x mol(-1), followed by HPS-3-B 1.113 x 10(5) g x mol(-1) and HPS-3-D 8.457 x 10(4) g x mol(-1) HPS-3-A showed the lowest weight average molecular weight of 1. 223 x 10(4) g x mol(-1) but the highest square radius of gyration, that is 55.5 nm. HPS-3-D had the widest range of molecular weight distribution in four components, with the polydispersity index (Mw/Mn) of 2.543. In the mobile phase, HPS-3-A was globular structure, HPS-3-C was random coil, HPS-3-B and HPS-3-D were both highly branched structure.

CONCLUSIONThe results provided necessary basis for further studies on molecular characteristics of the four components contained in HPS-3 and their relationship with bioactivity.

Chromatography, Gel ; Drugs, Chinese Herbal ; chemistry ; Lasers ; Light ; Polysaccharides ; chemistry ; Scattering, Radiation

Background and Objectives: Mesenchymal stem cells (MSCs) have immunomodulatory function and participate in the pathogenesis of many immunoregulation-related diseases, including psoriasis. Previously, we found that MSCs from psoriatic lesions overexpress the proinflammatory microRNA, miR-155 and exhibit a decreased immunosuppressive capacity. But the origin of these aberrant characteristics is still not clear. To investigate whether inflammatory cytokines in serum and peripheral blood mononuclear cells (PBMCs) from psoriatic patients can regulate the expression patterns of immunoregulation-related cytokines and the immunoregulation function of MSCs. Methods: and Results: Normal dermal mesenchymal stem cells (nDMSCs) were treated with serum or PBMCs derived from patients with psoriasis or healthy donors. Expression of miR-155 and immunoregulation-related genes in each MSCs were measured using real-time PCR or western-blot. Meanwhile, the immunosuppressive capacity of DMSCs was evaluated by its inhibitory ability on proliferation of activated PBMCs. Compared to control serum, psoriatic serum significantly increased the expression levels of miR-155 (27.19±2.40 vs. 3.51±1.19, p<0.001), while decreased TAB2 expression (0.28±0.04 vs. 0.72±0.20, p<0.01) in DMSCs. Expression levels of immunoregulation-related genes such as PGE2, IL-10, and TLR4 were also markedly down-regulated following the psoriatic serum treatment. Those DMSCs treated with healthy serum could inhibit PBMC proliferation, while those psoriatic serum-treated DMSCs could not inhibit PBMC proliferation effectively. Conclusions Psoriatic serum up-regulate the expression of miR-155, down-regulate the expression of immunoregulation- related genes (PGE2, IL-10, and TLR4) in DMSCs, and along with the inhibition of the immunosuppressive function of MSCs.