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OBJECTIVE To study the chemical constituents in S henfu injection and the anti-inflammatory activities of its polyacetylene compounds. METHODS Shenfu injection was separated and purified by macroporous adsorption resin ，medium pressure liquid chromatography ，preparative thin layer chromatography and reversed-phase semi-preparative high-performance liquid chromatography，and the compound structure was identified according to the physicochemical properties and spectral data. RAW 264.7 cell inflammation model was used to evaluate the anti-inflammatory activities of polyacetylene compounds . The effects of active polyacetylene compounds on the expressions of cyclooxygenase- 2（COX-2）protein were evaluated by Western blot assay. RESULTS Twelves compounds were isolated and identified from Shenfu injection ，including 8 ginsenoside compounds ，i.e. ginsenoside Rg 1（1），ginsenoside Re （2），ginsenoside Rb 1（3），ginsenoside Rk 1（4），20（R）-ginsenoside Rh 1（5），20（S）-ginsenoside Rg3 （6），notoginsenoside R 1（7），panaxatriol（8）；4 polyacetylene compounds ，i.e.（3R，9R，10R）-panaxytriol（9），panaxydol（10）， heptadeca-1，8-dien-4，6-diyne-3，10-diol（11）and panaxynol （12）. Among 4 polyacetylene compounds ，only compound 10 had anti-inflammatory activity. Compound 10 was not toxic to normal RAW 264.7 cells；when the concentration of compound 10 ranged 12.5-50.0 μmol/L，it could significantly reverse the lipopolysaccharide-induced NO content increase in cell supernatant （P＜0.05 or P＜0.01）；when the concentration of co mpound 10 was 50.0 μmol/L，it could significantly reverse the lipopolysaccharide-induced protein expression increase of COX- 2 in cells （P＜0.05）. CONCLUSIONS Compounds 4，7，10-12 are identified and reported in Shenfu injection for the first time ，and panaxydol possesses a certain anti-inflammatory effect.
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OBJECTIVE：To establish an HPLC fing erprint of Ginsen g Radix et Rhizoma Rubra ，and to optimize its processing technology. METHODS ：HPLC method was adopted. The determination was performed on Waters SymmetryShield TM RP18 column with mobile phase consisted of acetonitrile-water （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃，and the detection wavelength was 203 nm. The sample size was 10 μL. Using ginsenoside Rb1 as reference peak ，HPLC fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra was established. The similarity of them was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint （2012 A edition ） to confirm common peak. With steaming temperature，time and drying method as factors ，using the content of ginsenoside and fingerprint similarity as index ，the processing technology was optimized with L 16（43）orthogonal test design and verified. Cluster analysis was conducted with SPSS 19.0 statistical software of 10 batches of Ginseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample. RESULTS ：There were a total of 13 common peaks in the fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra. The similarity was more than 0.920；3 common peaks were identified ，such as ginsenoside Rg 1，ginsenoside Re ，ginsenoside Rb 1. The optimal processing technology included that steamed at 100 ℃ for 150 min，dried at 60 ℃. The results of validation test show that the contents of ginsenoside Rg 1，Re and Rb 1 were 0.26％-0.29％，0.17％-0.20％，0.47％-0.54％，and the similarity between 3 batches of Ginseng Radix et Rhizome Rubra optimal processed sample and the control fingerprints was more than 0.970. The results of cluster analysis showed that 10 batches of Gimseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample could be clustered into two categories；HS3-HS10 could be clustered into one category ，and 3 batches of optimal processed sample ，HS1 and HS 2 be clustered into one category. CONCLUSIONS ：Established fingerprint can be used for the optimization of processing technology of Gimseng Radix et Rhizoma Rubra ，and characterize the correlation between f luctuation of technology parameter and quality of medicinal material；the optimal processing technology is reasonable an d
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Objective To establish the methods of microbial limit test for three kinds of preparations with heavy metals, such asDingxian Pills,Pizhi Lotion andJiawei Huangqin Ointment.Methods According to Chinese Pharmacopoeia 2010, the recovery rates of bacteria, fungus and yeast treated by three preparations were detected. And the methods of testing control bacteria were also validated.Results Culture medium dilution method was proved to be applicable forDingxian Pills. Culture medium dilution method combined with pre-filtration method was proper for Pizhi Lotion. And the extraction method was adopted forJiawei Huangqin Ointment. The recovery rates of these five validation strains reached 70% by appropriate methods. And the same methods were used for validation of the control bacteria.Conclusion The methods of microbial limit test for these three different preparations were established through this study.
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OBJECTIVE:To establish the method for determination of baicalin in Chang’an capsule by RP-HPLC. METHODS:HPLC was performed on C 18 column with methanol-0.07%phosphoric acid solution(44∶56)as mobile phase at a flow rate of lml/min and the temperature of column kept at room temperature.The UV detection wavelength was280nm.RESULTS:The linear range of baicalin was sample size0.1054?g～1.0540?g(r=0.9996),the average recovery was97.44%(RSD=2.92%,n=5).CONCLUSION:The method was simple,accurate and fast,and can be used for the determination of baicalin.