Content Determination of Saccharide in Polysaccharides Containing Galacturonic Acid by Phenol-sulfuric Acid Method Combined with Calibration Factor Method / 中国药房

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.

Study on Producing and Preparing Integration Process based on FreshCodonopsis pilosula Nannf.var.modesta (Nannf.) L.T.Shen / 中国中医药信息杂志

Objective To study the effects of different drying processes on the effective constituents in Wen Codonopsis pilosula (WPP) Decoction Pieces; To develop the optimized producing and preparing integration process based on fresh WPP.MethodsFresh WPP in harvest period was prepared respectively as follows:① Fresh WPP was dried to different percentage (30%–100%) of original moisture contents of crude drugs at 80℃ in oven, then sliced and dried at 50℃ to obtain eight decoction pieces of WPP (XⅠYP1–8).② The fresh WPP was baked to 50% of water content of crude drug under different temperatures (50–120℃), respectively, then sliced and dried at 50℃ to obtain eight decoction pieces of WPP (XⅡYP1–8).③ Dried WPP was moistened, sliced and naturally dried, then were renamed as traditional decoction pieces. The contents of lobetyolin and atractylenolideⅢ was determined by HPLC. The phenol-sulfuric acid and colorimetric method were applied respectively to detect contents of polysaccharide and total flavonoids. The contents of aqueous and alcoholic extracts were determined simultaneously. Results The contents of alcoholic extracts (55.36%), aqueous extracts (54.91%) and atractylenolideⅢ (10.95 μg/g) in XⅠYP3 pieces were higher than other decoction pieces.Conclusion The optimized process was that fresh WPP was baked to water content of 50% at 80℃, then sliced and dried. Compared with conventional preparing methods,the integration process was time-saving and effort-saving. Meanwhile, the prepared pieces have higher content of active ingredients.

Study on molecular characteristics of four components contained in Hedysari Radix polysaccharide by gel permeation chromatography-multiangle laser light scattering technology (GPC-MALLS) / 中国中药杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>The 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.</p><p><b>RESULT</b>Among 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.</p><p><b>CONCLUSION</b>The results provided necessary basis for further studies on molecular characteristics of the four components contained in HPS-3 and their relationship with bioactivity.</p>

Phamacokinetic study of bergapten in rats plasma by LC-MS/MS / 中国中药杂志

<p><b>OBJECTIVE</b>To determine bergapten's concentration in plasma and observe its pharmacokinetics in rats using a combined LC-MS/MS analytical method.</p><p><b>METHOD</b>Blood samples were separated on a Hypersil ODS column (4.6 mm x 250 mm, 5 mm) at a temperature of 30 degrees C, and mobile phase consisted of water and methanol (22.5: 77.5) at a flow rate of 0.8 mL x min(-1).</p><p><b>RESULT</b>The methodological study showed a good linear relationship of 8.12-162.4 g x L(-1) (r = 0.9999). The inner and inter-days relative standard deviations were both less than 10% , indicating legitimate precise and accuracy to the requirement of biological sample analysis.</p><p><b>CONCLUSION</b>The method is suitable for in vivo quantitative analysis for bergapten due to its accuracy, sensitivity and specificity. The pharmacokinetic process in rats forms a two-compartment model with first-order absorption.</p>

Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography coupled with diode array detector and evaporative light scattering detector / 药物分析学报

A simple and accurate high-performance liquid chromatography（HPLC）coupled with diode array detector（DAD）and evaporative light scattering detector（ELSD）was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation（ZFP）.The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis（HCA）,which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography coupled with diode array detector and evaporative light scattering detector / 药物分析学报

A simple and accurate high-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) and evaporative light scattering detector (ELSD) was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation (ZFP).The monitoring wavelengths were 254,275 and 328 nm.Under the optimum conditions,good separation was achieved,and the assay was fully validated in respect of precision,repeatability and accuracy.The proposed method was successfully applied to quantify the six ingredients in 31 batches of ZFP samples and evaluate the variation by hierarchical cluster analysis (HCA),which demonstrated significant variations on the content of these compounds in the samples from different manufacturers with different preparation procedures.The developed HPLC method can be used as a valid analytical method to evaluate the intrinsic quality of this preparation.

Evaluation preparation technology of Xiaochaihu granules using fingerprint-peak pattern matching / 药物分析学报

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.

Simultaneous determination of five active components in compound preparation of Dangguishuan by high-performance liquid chromatography / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a HPLC method for determination of matrine, oxymatrine; ferulic acid, L-shikonin and beta, beta-dimethylacrylshikonin in compound preparatioti of Dangguishuan.</p><p><b>METHOD</b>The 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.</p><p><b>RESULT</b>All 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%.</p><p><b>CONCLUSION</b>The 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.</p>

Coumarins from Peucedanum harry-smithii var. subglabrum / 中国中药杂志

The root of Peucedanum harry-smithii var. subglabrum was extracted with methanol, then separated with solvents at different polarity into four fractions: aqueous (H2O), ethyl acetate (AcOEt), chloroform (CHCl3) and petroleum ether (DAB-6). From AcOEt psoralen, bargapten, xanthotoxin, marmesin, umbelliferone, scopoletin, (+/-) peuformosin, Pd-I b, (+/-) selinidin, praeruptorin D were isolated by column chromatography on silica gel, using petroleum ether/ethyl acetate as eluent. The structures of the coumarins were identified by 1H-NMR and 13C-NMR.

Determination of polysaccaride content in Radix Hedysari by HPLC / 中成药

AIM: To establish a method for determing polysaccaride content of Radix Hedysari(Hedysarum Polybotrys Hand-Mazz). METHODS: To determine content of Radix Hedysari polysaccaride by HPLC and ELSD. RESULTS: The composes of Radix Hedysari polysaccaride Ⅰ、Ⅱ、Ⅲ、Ⅳ are all Rhamnose、Arabinose、Xylose、Glucose、Galactose,the average content of polysaccharides is 25.34%、24.23%、15.08%、17.24%. CONCLUSION: The contents of Radix Hedysari polysaccaride in Radix Hedysari can be determined by HPLC.

Chromatography fingerprint of Baitiao Radix Codonopsis in Gansu Province by HPLC / 中成药

AIM To establish chromatography fingerprint of Radix Codonopsis. METHODS HPLC was applyed on a Hypersil ODS column with water-methanol gradient elution, flow-rate of 0.8 ml?min -1 . Determination of content of atractylenodie Ⅲ adopted ELSD and PAD with methanol-water(67∶ 33), flow-rate of 1.0 ml?min -1 , column temperature at 30?C , ELSD:carried-gas flow-rate of 2.2 L?min -1 , drift-tuber temperature was 80?C ;PAD:wavelength was 220 nm. RESULTS 25 common peaks were picked up, their similarity among 10 batchs of samples was more than 98.12 % based on marker composition(Atractylenolide Ⅲ). CONCLUSION The chromatography fingerprint of Baitiao Radix Codonopsis could be used as their characteristic chromatography fingerprint of hydrophobic fraction.

Determination of formononetin and calycosin from Zhengqifuzheng Preparation by HPLC / 中成药

AIM: To establish a method of RP-HPLC for determination of calycosin and formononetin in Zhengqifuzheng Preparation(Radix Astragali, Fructus Ligustri Lucidi, etc.). METHODS: Kromasil ODS column(5?m,4.6mm?250mm) was used to determine two isoflavoniods with water-methanol gradient elution, at column temperature of 25℃, flow-rate of 1.0mL?min -1 and detection wavelength at 254nm. RESULTS: The average recoveries of formononetin and calycosin in Zhengqifuzheng Capsules were 96.53% and 98.34% and RSD were 2.25% and 4.01%, respectively, the average recoveries of formonoenetin and calycosin in Zhengqifuzheng Granules were 97.20% and 95.65% and RSD were 2.66% and 2.32%, respectively. CONCLUSION: The method is sample, reliable, accurate, practical, and can provid a new idea to study the quality standard of this preparation.