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Objective: To study on changes of active components in Lianzhifan solution (LZF) under different storage conditions, which may provide a reference for the application and quality improvement of the LZF. Methods: HPLC analysis was performed on Eclipse Agilent C18 column (250 mm × 4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 238 nm, and the column temperature was 25 ℃. A fingerprint analysis method for the chemical composition of LZF was established to analyze the change of main active components with time under sealed and unsealed storage conditions. Results: The precision, stability, repeatability and sample recovery of the HPLC fingerprint of LZF were all in accordance with the requirements, and can be used for the analysis of the chemical constituents of LZF. The similarity values of sample fingerprints and control fingerprints at different time points under both storage conditions were greater than 0.95; The nine chemical components in LZF were identified by using the external labeling method and spectral characteristic analysis. The content of alkaloids (columbamine, epiberberine, jatrorrhizine chloride, coptisine chloride, palmatine chloride, and berberine) did not change significantly under the two storage conditions. However, the content of the iridoids, genipin-1-β-D-gentiobioside and geniposide was significantly reduced, and the content of genipin was increased in the first 3 months but decreased after 3 months. It is speculated that genipin-1-β-D-gentiobioside and geniposide can be converted to genipin by the enzyme in solution, and genipin may be degraded by other enzymes produced by microorganisms. Conclusion: During the six-month storage period, LZF still has a convert tendency, which may be caused by the action of enzymes produced by microbes. In order to improve the quality consistency of LZF product, it is recommended to adopt more scientific method to judge the endpoint and it is better to inactivate enzyme and sterilize after the end of the fermentation processing.
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Objective To establish a method for determining five isoquinoline alkaloids (including berberine,jatrorrhizine,jatrorrhizind,coptisine and palmatine) in Erhuang Xiaoyan tablets simultaneously.Methods Determination by reversed phase high performance liquid chromatography (RP-HPLC).Column:YMC-Pack ODS-AM (4.6 mm× 250.0 mm,5 μm);mobile phase:elution with 0.1％ (v/v) formic acid aqueous solution-acetonitrile mobile phase gradient;flow rate:0.4 mL/min;injection volume:10 μL;detection wavelength:345 nm;column temperature:27 ℃.Results Five of isoquinoline alkaloids in Erhuang Xiaoyan tablets were separated perfectly.The good linear relationships were obtained in the following ranges,1.526 4-5.342 4μg (r=0.999 9) for berberine,0.403 2-1.411 2 μg (r=0.999 6) for palmatine,0.309 7-1.083 9 μg (r=0.999 8) for coptisine,0.111 2-0.389 2 μg (r=0.999 8) for jatrorrhizine and 0.162 2-0.567 7 μg (r=0.999 6) for epiberberine.The recoveries were 98.69％,98.66％,98.67％,98.67％ and 98.67％,respectively.Conclusion The method possesses high feasibility,strong specificity,high accuracy and good reproducibility,which can be used for the quality control of Erhuang Xiaoyan tablets.
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Objective: To explore the anti-inflammatory mechanism of Scutellarlae Radix (SR) by the network pharmacology. Methods: Firstly, the components in SR were searched through TCMSP database and screened with “Lipinski rule” and “Oral Bioavailability > 30%” rules. The targets of above components selected by PharmMapper web server and Cytoscape 3.4.0 was used to build a network between components and targets (component-target network, CTN). Secondly, “anti-inflammatory” targets was searched from Therapeutic Target Database (TTD) with keyword “anti-inflammatory”, and targets retrieved were used to build a protein-protein interaction (PPI) network based on the analysis by String database. To obtain anti-inflammatory targets of the active components in SR, the PPI network was fused with the CTN. Finally, the DAVID database was used to perform KEGG pathway enrichment analysis in order to explore the anti-inflammatory mechanism of SR. Results: Twenty-eight components in SR were obtained, including flavonoids such as baicalin, baicalein, wogonin, wogonoside, etc, alkaloids such as berberine, and epiberberine, and phenols such as dihydromyricetin, etc. Mitogen-activated protein kinase (MAPK14), tumor necrosis factor receptor superfamily 1A (TNFRSF1A), epidermal growth factor receptor (EGFR), and E-selectin (SELE) were the main targets of SR' anti-inflammatory effect. Salvigenin, epicatechin, and astragalusine mainly acted on MAPK14; Carthamidin acted on TNFRSF1A; Dihydromubutin A, 5,7,4’-trihydroxy-8-methoxyflavone, 5,7,4’-trihydroxy-8-methoxyflavanone, baicalin, and other components mainly acted on EGFR. There were 11 KEGG pathways, mainly related to TNF signaling pathways, MAPK signaling pathway, etc. Conclusion: There are three main anti-inflammatory mechanisms in SR, which can inhibit the production of inflammatory factors, inhibit the binding of inflammatory factors to their respective receptors, and block the initiation of inflammatory reactions.
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AIM To establish an HPLC method for the simultaneous content determination of six constituents in Jiaotai Pills (Coptidis Rhizoma and Cinnamomi Cortex).METHODS The analysis of 30％ methanol of this drug was performed on a 30 ℃ Agilent ZORBAX SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-KH2PO4flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelength was set at 276 nm.RESULTS Epiberberine,jatrorrhizine hydrochloride,coptisine hydrochloride,palmatine chloride,berberine hydrochloride and cinnamaldehyde showed good linear relationships within the ranges of 0.64-41.24 μg/mL (R2 =0.999 9),0.65-43.76 μg/mL (R2 =1.000 0),0.82-52.65 μg/mL (R2 =0.999 9),0.79-50.70 μg/mL (R2 =0.999 9),3.08-197.20 μg/mL (R2=0.999 8) and 0.65-41.65 μg/mL (R2 =0.999 9),whose average recoveries were 98.06％,102.76％,99.27％,99.75％,96.74％ and 101.33％ with the RSDs of 0.56％,0.54％,0.39％,0.55％,0.48％ and 2.14％,respectively.CONCLUSION This accurate,sensitive,stable and reproducible method can be used for the quality control of Jiaotai Pills.
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Objective: To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of six alkaloids in crude and processed Coptidis Rhizoma. Methods: An HPLC method was established to determine the six alkaloids (jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, and berberine hydrochloride) by the external standard method (ESM). With this HPLC method, the berberine hydrochloride was used as the internal standard (IS) to determine five relative correction factors (RCFs) of the five other alkaloids, and their contents in all samples were calculated by their RCFs at the same time. Compared with the content results determined by the ESM and QAMS, the feasibility and accuracy of QAMS method were verified. Results: Within a certain range, the RCFs of jatrorrhizine hydrochloride, columbamine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride, and palmatine hydrochloride to berberine hydrochloride were 1.131, 0.999, 1.011, 1.076, and 1.025, respectively, with the good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. Conclusion: The QAMS method is feasible and accurate for the simultaneous determination of the six alkaloids in crude and processed Coptidis Rhizoma.
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Objective: To establish and study the UPLC-MS/MS fingerprint, to ascertain the medicinal source of chromatographic peaks, and to identify the chemical constituents for providing scientific basis in effective substance foundation and quality control of Wuji Pill. Methods: The fingerprint of Wuji Pill was developed with ultra-performance liquid chromatography (UPLC), and the Acquity UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm) was used in the gradient elution with a mobile phase of acetonitrile-water (0.05% formic acid): The flow rate was 0.4 mL/min, the column temperature was 40℃. Similarity evaluation was used to evaluate the quality of herbs from different areas. Results: The fingerprint of Wuji Pill was established with good precision, reproducibility, and stability obtaining within 55 min, and 38 peaks in the fingerprint were designed. Twenty samples could be classified into two clusters. According to the reference, and qualitative and references identified a total of nine peaks were berberine, palmatine, jatrorrhizine, coptisine, epiberberine, evodiamine, rutaecarpine, evodine, and paeoniflorin inferred that the four common peaks were magnolia base magnoflorine, albiflorin, theophylline, and methyl-2-nonyl-4(1H)-quinolone. Conclusion: The establishment of UPLC fingerprint of Wuji Pill and application of chemical pattern recognition can provide a more comprehensive reference for the quality control of herbs.
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Everted intestinal sac models were used to investigate the intestinal absorption of the 4 alkaloids(berberine, palmatine, coptisine, and epiberberine) in Fuzheng Xiaozheng Fang(FZ) at different intestine segments. The absorption parameters of each component were calculated; SPSS 20.0 software was used to analyze the data and evaluate the absorption characteristics at different intestinal segments. The results showed that all the four active ingredients conformed to zero-order absorption rate. There was significant difference in absorption rate constant (Ka) between the four ingredients at low dose and medium and high dose groups(P<0.05), but there was no significant difference in Ka between medium dose and high dose. The absorption mechanization of four ingredients presented two absorption manners： positive diffusion and passive absorption. The absorptive amount of 4 alkaloids in ileum was slightly greater than that of jejunum, but no significant differences were observed, which indicated that these four alkaloids had no specific absorption windows in intestinal segment.
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Objective To build a method for determination of berberine, epiberberine, coptisine and palmatine in Shutongantai Capsules by HPLC.Methods Thermo C18 column (5μm, 4.0 mm× 250 mm) was used with acetonitrile-0.05 mol/L potassium dihydrogen phosphate solution (50∶50, 0.4 g sodium dodecyl sulfate was added in each 100 mL solution, to adjust the pH 4.0 with phosphoric acid) as mobile phase;the flow rate was 1.0 mL/min;detection wavelength was 345 nm;column temperature was 30℃.Results Berberine hydrochloride showed a good linear relationship in the range of 0.180 34-0.901 68μg (r=0.999 8).Conclusion The method is simple, accurate and reproducible, and can be used for the quality control of Shutongantai Capsules.
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Objective: To investigate the inhibition of rutaecarpine, a main component in Evodiae Fructus, on the hepatic metabolism of five Coptis alkaloids, and to provide the basis for further study of compatibility mechanism between Coptidis Rhizoma and Evodiae Fructus. Methods: Using rat liver microsome incubation method, the inhibition of rutaecarpine on hepatic metabolism of five Coptis alkaloids in vitro was investigated. Results: Rutaecarpine could inhibit the in vitro hepatic metabolisms of coptisine, epiberberine, berberine, palmatine, and jatrorrhizine. The half inhibitory concentration (IC50) was all greater than 50 μmol/L which showed rutaecarpine had a weak inhibition on Coptis alkaloids. The differences of inhibition constant (Ki) were statistically significant (P jatrorrhizine > palmatine > epiberberine > coptisine. Conclusion: The results could provide the basis to learn the major role on the links that Evodiae Fructus acted on Coptis alkaloids and reveal the compatibility mechanism between Coptidis Rhizoma and Evodiae Fructus.