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ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion（SCU-PC-NE）， such as release， cell uptake and tissue distribution， and to investigate its effect on ameliorating lipopolysaccharide（LPS）-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC， ethyl oleate， Kolliphor HS15， 1，2-propylene glycol（50， 400， 514.3， 85.7 mg）， respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope， the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis， thiazolyl blue（MTT） colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells（HUVECs）， the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe， the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS（1 mg·L-1） and HUVECs， the effect of SCU-PC-NE on the levels of interleukin（IL）-1β and IL-6 were determined by enzyme-linked immunosorbent assay（ELISA）， 18 Kunming male mice were randomly divided into blank group， model group， blank preparation group（equivalent to high dose group）， SCU group and SCU-PC-NE low and high dose groups（5， 10 mg·kg-1）， 3 mice in each group， and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h， equal volume of normal saline was given to the blank group and the model group， and the drug was administered for 4 consecutive times. Except for the blank group， the endothelial inflammatory injury was induced by intraperitoneal injection of LPS（10 mg·kg-1） at 12 h before the last administration in each group. Hematoxylin-eosin（HE） staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light， mostly spherical with rounded appearance and relatively uniform particle size distribution， with the average particle size of 35.31 nm， Zeta potential of 7.23 mV， and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group（P<0.01）. Compared with normal mice， the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen， kidney， brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS（P<0.05， P<0.01）， especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group（P<0.05， P<0.01）， and compare with the model group， all administration groups significantly down-regulated IL-1β level， with the strongest effect in the SCU-PC-NE high-dose group（P<0.01）， and all administration groups significantly down-regulated IL-6 level， with the strongest effect in the SCU-PC-NE low-dose group（P<0.05）. Compare with the blank group， the results of HE staining showed that the endothelial cells were damaged， the elastic fibers were broken and arranged loosely in the model group， although similar vascular injury could be observed in the blank preparation group， SCU group and SCU-PC-NE low-dose group， the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE， which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS， suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.
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OBJECTIVE：To optimi ze and improve the quality standard for Keqing capsules. METHODS ：According to general rule 0502 method stated in 2015 edition of Chinese Pharmacopeia （part Ⅳ），TLC method was used to identify Reineckia carnea and Morus alba in Keqing capsules [the developing solvents were dichloromethane-ethyl acetate-formic acid （10 ∶ 4 ∶ 0.2，V/V/V） and ethyl acetate-carbinol-ammonia （12 ∶ 2 ∶ 1，V/V/V），respectively]. The contents of morphine and codeine phosphate in Keqing capsules were determined by HPLC. The determination was performed on XBridge C 18 column with mobile phase consisted of acetonitrile-0.01 mol/L potassium dihydrogen phosphate aqueous solution （pH value adjusted to 2.7 with 5％ phosphoric acid solution）（5 ∶ 95，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm，and the column temperature was 35 ℃. The sample size was 10 µL. RESULTS ：In TLC of R. carnea and M. alba in samples ，same color spots were shown in the correspon ding positions of reference substance chromatogram without interference from negative control. The linear range of morphine and codeine phosphate were batches of Keqing capsules were 0.97-1.37，0.16-0.37 mg/g，respectively. CONCLUSIONS ：TLC identification method for R. carnea and M. alba ，as well as HPLC content determination method for morphine and codeine phosphate in Keqing capsules are established；the method is simple ，accurate and reliable with strong specificity ，which improves the quality standard of Keqing capsules.
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AIM To establish the quality standard for Cyclocarya paliurus (Batal.) ljinsk..METHODS The contents of water,ash and extract in twelve batches of samples were determined.TLC and HPLC were adopted in the qualitative identification and quantitative determination of quercetin and kaempferol,respectively,and phenol-sulfuric acid method was used for the polysaccharide content assessment.RESULTS The average contents of water,total ash,acid-insoluble ash,water-soluble extract and alcohol-soluble extract were 11.05％,5.81％,1.70％,11.25％ and 10.16％,respectively.The clear TLC spots demonstrated their strong specificity.Quercetin and kaempferol showed good linear relationships within the ranges of 0.004 953-0.022 29 mg/mL and 0.005 748-0.028 74 mg/mL (r =0.999 9),whose average recoveries were 97.1％ (RSD =2.59％) and 97.9％ (RSD =2.86％),respectively.Polysaccharide showed a good linear relationship within the range of 0.021 22-0.095 58 mg/mL,whose average recovery was 97.2％ (RSD =2.42％).The contents of three constituents in various batches of samples showed obvious differences.CONCLUSION In C.paliurus,the contents of water,total ash,acid-insoluble ash,water-soluble extract and ethanol-soluble extract should not be more than 13.0％,7.0％,2.0％,13.5％ and 12.0％,while those of quercetin,kaempferol and polysaccharide (calculated by dry product) should not be less than 0.040％,0.070％ and 0.60％,respectively.
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Objective To establish a methodological study pattern of quantitative analysis of multi-components by single marker ( QAMS) , and examine its feasibility and technical applicability in the quality control of compound preparation of traditional Chinese medicine-Jinhoujian spray. Methods Gas chromatographic method ( GC) was used and naphthalene served as the internal standard. Menthol was used as the reference substance. The relative correlation factors ( RCF) of 1,8-Cineole, camphor and borneol to menthol were calculated and established to carry out QAMS.The accuracy of this method was confirmed by comparison of internal standard method. Results The reproducibility of relative correction factor was perfect. The two methods did not show significant difference in 10 bathes of samples. Conclusion The QAMS method is feasible, credible, and can be used to determine active ingredients in Jinhoujian spray.
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OBJECTIVE:To establish an HPLC method for the determination of Diosgenin in Bixiefenqing pills.METHODS:The samples were separated on DiamonsilTM C18(200 mm?4.6 mm,5 ?m)column.The mobile phase consists of acetonitrile-water(92∶8) at a flow rate of 1.0 mL?min-1.The detecting wavelength was set at 203 nm and the column temperature was set at 35 ℃.RESULTS:The linear range of Diosgenin was 0.115 36～1.038 24 ?g(r=0.999 9).The average recovery was 98.71%(RSD=1.92%,n=9).CONCLUSION:The method was proved to be simple and accurate,and suitable for the identification and quality control of Bixiefenqing pills.
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Objective: To establish the determination method of gallic acid in Ninbitai Capsules.Methods: HPLC with Shimadzu Shim-pack VP-ODS(250mm?4.6mm,5??m) column was used. Methanol-water-dimethylformamide-acetic acid glacial (1∶81∶15∶3) was used as mobile phase. The flow rate was 0.5mL/min. The detection wavelength was at 272nm.Results: The linearity of this method was well. The recovery of the added sample was 98.16%. RSD was 1.18%.Conclusion: This method is convient with a good separating degree and can be used for the quality control of Ninbitai Capsules.