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OBJECTIVE：To study the effects and potential mechani sm of deoxyschizandrin on the proliferation ，migration and invasion of nasopharyngeal carcinoma cell HONE- 1. METHODS ：HONE-1 cell was set as cell model ，while CCK- 8 test，wound healing assay and Transwell chamber test were used to detect the proliferation ，migration and invasion ability changes of HONE- 1 cells after treatment with different concentrations [ 0（blank control ），10，20，40 μmol/L] of deoxyschizandrin. Computer molecular docking was performed to analyze the binding ability between deoxyschizandrin and Met protein. Western blotting assay was used to detect the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells. RESULTS ：Compared with blank control ，the proliferation ，migration and invasion ability of cells after treated with 10，20，40 μmol/L deoxyschizandrin were all decreased significantly （P＜0.05）. Results of molecular docking revealed that deoxyschizandrin could stably bind with the activity pocket of Met protein. Results of Western blotting assay demonstrated that compared with blank control ，10，20，40 μmol/L deoxyschizandrin all decreased the relative protein expressions of p-Met ，p-PI3K，p-Akt，Bcl-2 and N-cadherin in cells significantly（P＜0.05）. CONCLUSIONS ：Deoxyschizandrin can inhibit the proliferation ，migration and invasion of HONE- 1 cell via inhibiting the activation of Met/PI 3K/Akt signaling pathway.
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Aim To investigate the synergy effects of deoxyschizandrin and gemcitabine (GEM) on the proliferation of HepG2 human hepatocellular carcinoma cells in vitro and the underlying mechanism. Methods CCK8 method and colony formation assay were used to detect the effects of deoxyschizandrin monotherapy, GEM monotherapy and combination therapy on the proliferation of HepG2 cells. Flow cytometry was used to detect the change of apoptosis rate of HepG2 cells after treatment with single drug or the combination use of two drugs. Western blot was performed to detect the expression of BCL2, BAX, pro-caspase3, caspase3, pro-caspase9, caspase9, ß-catenin and TCF-4. Results Deoxyschizandrin, GEM and combination group significantly inhibited the proliferation of HepG2 cells and promoted cell apoptosis. The effects of the combination group on HepG2 cells were significantly stronger than those of single-drug groups (P < 0. 05). Western blot results showed the expression of pro-caspase3 and pro-caspase9 was changed slightly within deoxyschizandrin, GEM and combination groups compared with that of normal control, while the expression of B C L 2, ß-catenin and TCF-4 protein expression was down-regulated significantly (P < 0. 05). The expression of B A X, cleaved-caspase3 and cleaved-caspase 9 protein increased significantly after treatment with deoxyschizandrin, GEM and combination group (P < 0. 05). Specially, the increasing effect of the expression of the protein in combined group was more significant than that of single drug groups (P < 0. 05). Conclusions The combination of deoxyschizandrin and GEM significantly inhibited the proliferation of HepG2 cells and induced cell apoptosis, as well as suppressed the ß-catenin/TCF-4 pathway.
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Objective: To establish HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneously determine the contents of the six constituents, acteoside, martynoside, calycosin 7-O-β- D-glucopyranoside, schisandrin, formononetin and deoxyschizan- drin. Methods: The analysis of 50% methanol extract of this drug was performed on a 30℃ thermostatic Waters Symmetry C18 column, with the mobile phase comprising of acetonitrile(A)-0.2% formic acid solution(B)flowing at 1.0 ml/min in a gradient elution manner. The detection wavelength was set at 330 nm for acteoside and martynoside, 254 nm for calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin. Results: There were thirteen common peaks in the fingerprints of ten batches of samples with the similarities more than 0.9. Acteoside, martynoside, calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin showed good linear relationship within the ranges of 1.47-36.75, 0.66-16.50, 0.89-22.25, 2.58-64.50, 1.91-47.75 and 0.77-19.25 μg/ml(r≥0.9993), and their average recoveries were 97.57%, 99.22%, 96.69%, 100.01%, 98.79% and 96.77%, with the RSD of 0.79%, 1.54%, 0.61%, 0.64%, 0.83% and 0.36%, respectively. Conclusion: The established method is easily operable and repeatable, which could be used for quality control of Qiwei Tangmaishu tablet.
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Objective: To explore the potential Q-markers between crude Schisandrae Chinensis Fructus (SCF) and vinegar-processed Schisandrae Chinensis Fructus (VSCF) based on multivariate statistical analysis and network pharmacology. Methods: UPLC-Q/TOF-MS was used to analyze the main lignans in SCF and VSCF, and the potential differences of chemical components (Q-markers) between SCF and VSCF were screened out by using multiple statistical methods. Furthermore, through network pharmacology and bioinformatics, the main action targets and pathways related to significantly different components were analyzed to construct the “component-target-pathway” network relationship and predict the potential quality markers between SCF and VSCF. Results: In this study, 40 different constituents of Schisandra chinensis between SCF and VSCF were screened, among which eight chemical markers had significant differences between SCF and VSCF. Five chemical constituents were identified and confirmed, namely 5-HMF, deoxyschizandrin and its isomer, schisandrin B, and schisantherin D. The other three chemical markers were speculated to be lignans by analyzing the first-and second-order mass spectrometry information. The results of network pharmacological analysis showed that the five potential quality markers identified were highly related to the main pharmacological effects of SCF. Finally, schisandrin B and 5-hydroxymethyl furfural were identified as the most representative potential quality markers. Conclusion: The results showed that the chemical composition of SCF had a series of complex changes. It was determined that schisandrin B and 5-hydroxymethyl furfural could be used as representative potential quality markers between SCF and VSCF. It is speculated that lignans may be the basis of the important effect of VSCF on liver protection.
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Objective To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components (deoxyschizandrin, γ-schizandrin, schizandrin, schizandrol B, schisantherin A, psoralen, isopsoralen, evodiamine, and rutaecarpine) in Sishen Pills. Methods The chromatographic separation was performed on an Agilent Zorbax Eclipse Plus C18 column (100 mm × 4.6mm, 3.5 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.4 mL/min, and the injection volume was 20 μL. The nine major bioactive components were detected using an electrospray ionization source in positive ionization mode (ESI+) and quantified by multiple reaction monitor (MRM) scanning at the same time. Results The linear ranges of deoxyschizandrin, γ-schizandrin, schizandrin, schizandrol B, schisantherin A, psoralen, isopsoralen, evodiamine, and rutaecarpine were 8.50-850.00 ng/mL (r = 0.999 7), 1.32-132.00 ng/mL (r = 0.997 4), 9.60-960.00 ng/mL (r = 0.999 8), 12.00-1 200.00 ng/mL (r = 0.999 3), 11.50-1 150.00 ng/mL (r = 0.997 9), 21.70-2 170.00 ng/mL (r = 0.999 7), 23.80-2 380.00 ng/mL (r = 0.999 6), 10.70-1 070.00 ng/mL (r = 0.999 5), 8.54-854.00 ng/mL (r = 0.998 0), and the average recoveries were 98.3% (RSD = 2.21%), 100.3% (RSD = 1.78%), 99.2% (RSD = 2.19%), 100.4% (RSD = 2.23%), 99.1% (RSD = 2.18%), 97.7% (RSD = 3.03%), 99.0% (RSD = 2.51%), 98.9% (RSD = 2.72%), and 100.3% (RSD = 2.10%), respectively. The contents of eight batches of the nine major bioactive components were 67.6-425.6, 0-131.5, 2.1-258.0, 0-71.2, 23.2-678.8, 806.4-1310.8, 718.5-1293.7, 11.5-123.2, and 10.9-62.4 μg/g, respectively. Conclusion The developed method is simple, specific, and sensitive, and it can be applied for the determination of nine major bioactive components and the quality control of Sishen Pills collected from different production batches.
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Objective To establish HPLC fingerprint strategy to simultaneously determine nine components in Liver-protection Agent, including loganin, paeonflorin, scutellarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid, and provide a scientific basis for the quality control of Liver-protection Agent and related medicinal preparations. Methods The analytical analysis was performed on an Agilent 1260-HPLC system equipped with a VWD detector and SB-C18 reversed phase column (Zorbax 150 mm × 4.6 mm, 5 μm). The analytes were eluted using a gradient mixture of two solvent: solvent A was distilled deionized water containing 0.1% ortho-phosphoric acid and solvent B was 100% acetonitrile. The mobile phase flow rate was 1.0 mL/min. The separation temperature was 30 ℃. The detection wavelengths were 236, 280, and 210 nm. The injection volume was 5 μL. Common patterns of HPLC fingerprints for 10 batches of Liver-protection Agent medicinal preparations were established, and chemometric analysis method was employed to analyze the hidden information. At the same time, methodological study was conducted for determinations of multiple components including loganin, paeonflorin, scutellarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid. Results The HPLC fingerprint strategy of Liver-protection Agent medicinal preparation had been set up, and 37 common peaks had been identified with the similarity of more than 0.9. Moreover, the samples were roughly divided into four categories by the methods of the systematic cluster analysis and principle component analysis. After validating the multiple component quantitative analysis condition through methodology, the average recovery rate was between 95.13% and 104.8%.The average mass concentration of loganin, paeonflorin, scutellarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid in ten batches of Liver-protection Agent was 216.3-223.0, 126.1-137.1, 144.7-149.0,1 623.1-1 992.7, 481.9-520.0, 14.9-18.7, 33.8-37.0, 2.9-3.7, and 39.7-43.6 mg/L, respectively. Conclusion The combination methods of HPLC fingerprint and simultaneous determinations of multiple components are rapid, simple, and reproducible, which can be adopted as one of the quality control methods for Liver-protection Agent and related medicinal preparations.
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AIM To establish an HPLC-DAD method for the simultaneous content determination of six constituents in Maiwei Dihuang Pills (Ophiopogonis Radix,Schisandrae chinensis Fructus,Rehmanniae Radix Praeparata,etc.).METHODS The analysis of 50％ methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent ZORBAX SB-C18column (4.6 mm × 150 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.2％ phosphate acid) flowing at 0.8 mL/min in a gradient elution manner,and the detection wavelengths were set at 220,230,236 and 274 nm.RESULTS Deoxyschizandrin,schizandrin B,schisandrin,paeoniflorin,paeonol and loganin showed good linear relationships within the ranges of 10-70,6.5-45.5,33.5-234.5,17-119,31-217 and 34-238 μg/mL (r ＞0.990 0),whose average recoveries (RSDs) were 99.6％ (1.7％),100.4％ (1.8％),100.7％ (1.8％),102.9％ (1.7％),102.2％ (1.5％) and 99.7％ (1.2％),respectively.CONCLUSION This simple and reproducible method can be used for the rapid quality control of Maiwei Dihuang Pills.
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This study was conducted to investigate the inhibitory effect and the molecular mechanism of deoxyschizandrin on the activity of NLRP3 (NOD-like receptor family, pyrin domain containing 3) inflammasome. Bone marrow-derived macrophages were used to study the effects of deoxyschizandrin on inflammasome activation using inflammasome inducers (ATP and nigericin). Cytotoxic effect was evaluated with CCK-8. The expression of IL-1β, caspase-1 in the supernatant and the expression of pro-caspase-1, pro-IL-1 β, ASC, NLRP3 in cell was detected by Western blot for the inhibitory effect of deoxyschizandrin (25, 50, 100 and 200 μmol·L-1) on the activity of NLRP3 inflammasome. Immunofluorescence was applied to investigate NF-κB (p65) transportation to the nucleus. The results of CCK-8 showed that the optimum concentration of deoxyschizandrin was 6.25-400 μmol·L-1. Deoxyschizandrin (25, 50, 100, and 200 μmol·L-1) could inhibit the activation of NLRP3 inflammasome caused by nigericin and ATP, and inhibit the secretion of IL-1 β, which was associated with inhibiting the cleavage of pro-caspase-1. The results of immunofluorescence and Western blot also suggest that the inhibitory activity of deoxyschizandrin on NLRP3 inflammasome was not dependent on NF-κB pathway and protein expression of NLRP3, ASC, pro-caspase-1 and pro-IL-1 β mediated by NF-κB. Our results confirmed that deoxyschizandrin could suppress the cleavage of pro-caspase-1 and inhibit the activity of NLRP3 inflammasome at 25-200 μmol·L-1 to reduce the inflammation response.
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Objective: To establish a method for HPLC determination of schisanthein, deoxyschizandrin and anwuligan in Wuzhi pellets simultaneously and control the quality of Wuzhi pellets.Methods: Schisantherin, deoxyschizandrin and anwuligan were determined by HPLC with a chromatographic column of Welch Ultimate XB-C 18 (150 mm× 4.6 mm , 3 μm), the mobile phase was tetramathylene oxide-water with gradient elution.The detection wavelength was 228 nm.The column temperature was 35℃.Results: The linear range of schisantherin was 0.040-0.805 μg (r=0.999 9),and the average recovery was 97.0% with RSD of 1.67%.The linear range of deoxychizandrin was 0.092-1.846μg (r=0.999 9),and the average recovery was 102.5% with RSD of 1.49%.The linear range of anwuligan was 0.020-0.998 μg (r=0.999 9),and the average recovery was 95.0% with RSD of 1.78% (n =6).Conclusion: The method is simple, accurate and reproducible.It can improve the quality standard of Wuzhi pellets.
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Objective To investigate the influence of deoxyschizandrin (Deo) on P-glycoprotein (P-gp).Methods The effect of P-gp on Deo (20,40,80 μg·mL-1) was studied in the Caco-2 cell model in vitro,and the apparent permeability coefficient (Papp) of Deo (20-160 μg·mL-1) on a P-gp substrate,rhodamine123 or cyclosporine A,was calculated.Healthy male Sprague-Dawley rats were randomly divided into five groups:blank control group,verapamil group,low-,medium-and high-dose Deo group (8 rats in each group).Rats in the low-,medium-and high-dose Deo group were intragastrically administered once daily with Deo at 8,16 and 32 mg·kg-1 for 3 consecutive days,while rats similarly received gavagewith verapamil (4 mg·kg-1) in the verapamil group and equal volume of purified water in the blank control group.Thirty minutes after the rats were treated with their respective drugs,rhodamine123 (5 mg· kg-1) was orally administrated.Then the pharmacokinetic profiles of rhodamine 123 were analyzed to evaluate the inhibitory ability of Deo on P-gp in vivo.Results The bidirectional transport rates of Deo (20,40,80 μg·mL-1) were similar,with non-selectivity.Deo (20-160 pg·mL-1)significantly inhibited the basolateral→apical(BL→AP) directional transports of rhodamine 123 and cyclosporine A in Caco-2 cell model (P ＜ 0.05) in a concentration-dependent manner.And Deo (8-32 mg· kg-1) also dose-dependently decreased the peak concentrations (Cm.) and the area under the plasma concentration-time curve (AUC0-t) of Rho123.Conclusion Deo can inhibit P-gp in vitro and in vivo,but it is not a P-gp substrate.
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Objective: To establish an HPLC fingerprint of the compounds in Qibai Pingfei Granules (QPG), and to make a quantitative analysis. Methods: Sample was extracted by 50% methanol. Phenomenex Luna C18 column (250 mm × 4.6 mm, 5 μm) was used with a mobile phase of methanol-0.2% formic acid gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 250 nm, and the column temperature was 30℃. The chemical component fingerprint similarity of 10 batches of QPG was calculated with Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012) published by National Pharmacopoeia Committee and the common peaks were identified by reference compounds. Results: Fingerprints of 10 batches of QPG were established and the similarities to the common mode were above 0.96. Totally 25 common peaks were found. Among them, peak 1 belonged to Pheretima, peaks 2 and 3 belonged to Pheretima and Ginseng Radix et Rhizoma (GRR), peak 4 belonged to GRR, Schisandrae Chinensis Fructus (SCF), and Chuanxiong Rhizoma (CR), peak 5 belonged to GRR, Allii Macrostemonis Bulbus (AMB), CR, and Pheretima, peaks 6, 8, 22, 23, 24, and 25 belonged to SCF, peak 7 belonged to Astragali Radix (AR) and SCF, peak 9 belonged to GRR and AMB, peak 10 belonged to Descurainiae Semen Lepidii Semen (DSLS) and AMB, peaks 11, 12, 13, 15, and 16 belonged to CR, peak 14 belonged to AR and DSLS, peaks 17, 18, 19, 20, and 21 belonged to AR. Based on the retention time, and UV absorption spectra of reference compounds, six constituents including caffeic acid (peak 12), ferulic acid (peak 13), schizandrol A (peak 22), schisantherin A (peak 23), deoxyschizandrin (peak 24), and schisandrin (peak 25) were identified. The linear ranges of caffeic acid, ferulic acid, schizandrol A, schisantherin A, deoxyschizandrin, and schisandrin were 3.38-108.02, 3.60-115.33, 2.99- 95.61, 2.81-89.77, 3.26-104.17, and 2.89-92.45 μg/mL, respectively. In 10 batches of QPG samples, the contents were as follows: caffeic acid of 0.412-0.429 mg/g, ferulic acid of 0.302-0.317 mg/g, schizandrol of A 0.182-0.195 mg/g, schisantherin A of 0.179-0.195 mg/g, deoxyschizandrin of 0.203-0.215 mg/g, and schisandrin of 0.131-0.144 mg/g, the amount of each indicator composition among different batches changed a litte, and the sample quality is stable. Conclusion: The method has good precision, reproducibile, stability, separation, and can be used for the quality control of QPG.
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Objective: To study the relativity between color and chemical composition in fruits of Schisandra chinensis by fruits' surface color value (L*, a*, and b*) and contents of active ingredients of fructus core S. chinensis). Methods: The surface chromatic value (multivariate linear regression equation of L*, a*, b* values and deoxyschizandrin, schizandrin B, schizandrol A, schinsantherin A) was established by SPSS 20.0 with mathematical statistics method, then the influence degree of the active ingredients of. S. chinensis was detected by F test. Results: The value of L* were significant negative with deoxyschizandrin (r ≥ 0.5), and other chromaticity values were very low or low-grade correlated with lignans Conclusion: The effective ingredient content of fruit of S. chinensis can be estimated by its chromatic values, and also can be used in the hierarchies of the fruit of S. chinensis, in order to provide better scientific basis of the quality of S. chinensis.
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Objective To establish a quantitative analysis of multi-component with a single-marker (QAMS) method for the quality control of Wuzi Yanzong Pills (WYP). Methods Six main effective components (schisandrin, hyperin, quercitrin, kaempferol 3-O-rutinoside, deoxyschizandrin, and γ-schizandrin) of WYP were simultaneously separated on a reversed-phase column (Ultimate LP-C18) with high-resolution of each chromatographic peak by high performance liquid chromatography (HPLC). Schisandrin was selected as the internal reference, and the relative correlation factors (RCFs) of other five components were calculated to achieve QAMS. The ruggedness of RCFs was tested on different instruments and columns. Moreover, results of the QAMS were compared with the external standard method. Results Within a certain linear range, the RCFs of hyperin, quercitrin, kaempferol 3-rutinoside, deoxyschizandrin, and γ-schizandrin were 0.36, 4.86, 0.88, 7.34, and 6.35, respectively. The repeatability was good under different experimental conditions. There were no significant differences between the calculated value and estimated value on QAMS and external standard method. Conclusion The QAMS method can be used to assay the content of six components of WYP simultaneously and control the quality of WYP simplely, reliably, and accurately.
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Objective:To study the inhibitory effect of deoxyschizandrin on the growth of brain glioma C6 cells, and to explore its mechanism.Methods:The rat glioma C6 cells were cultured and divided into control group,50, 100,and 200 mg·L-1 deoxyschizandrin groups.The proliferation rates of C6 cells were examined by MTT assay;the changes of cell cycles were examined by flow cytometry;the expression levels of CyclinD1,Bax,Bcl-2 and Caspase-3 proteins in supernant were detected by ELISA assay. Results:Compared with control group, the proliferation rates at 24 and 48 h in 50,100,and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P <0.01),and the proliferation rates at 72 h in 100 and 200 mg·L-1 deoxyschizandrin groups were significantly decreased (P < 0.05 or P < 0.01 ). Compared with control group, the percentage of cells at SubG1 phase in 200 mg·L-1 deoxyschizandrin group was increased (P < 0.05 ), and the percentage of cells at S phase was decreased (P <0.05).Compared with control group,the expression levels of CyclinD1 in 100 and 200 mg· L-1 deoxyschizandrin groups were decreased (P < 0.01 );the expression levels of Bax protein in deoxyschizandrin groups were increased (P < 0.05 or P < 0.01 ), and the expression level of Bcl-2 protein in 200 mg · L-1 deoxyschizandrin group was decreased (P < 0.01 ), and the Bax/Bcl-2 value in deoxyschizandrin groups were increased (P < 0.01 ); the expression level of Caspase-3 protein in 200 mg · L-1 deoxyschizandrin group was increased (P < 0.01 ).Conclusion:Deoxyschizandrin could inhibit the growth of glioma cells through down-regulating the expression levels of CyclinD1 protein and up-regulating the expression levels apoptotic factors Bax and Bcl-2.
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Objective:To establish an HPLC method for the simultaneous determination of adenosine, schisandrin, schisantherin,deoxyschizandrin and schisandrin B in Hugan tablets. Methods:An Agilent Eclipse XDB column(250 mm × 4. 6 mm,5 μm)was used with the mobile phase of acetonitrile- 0. 7% phosphoric acid solution with gradient elution. The flow rate was 0. 80 ml·min -1 . The detection wavelength was set at 260 nm in 0-14 min and 250 nm in 14-58 min,the column temperature was 30℃ ,and the sample size was 10 μl. Results:There was a good linear relationship within the range of 2. 35- 47. 00 μg· ml -1(r = 0. 999 8)for adenosine,14. 90-297. 00 μg·ml -1(r = 1. 000 0)for schisandrin,3. 46- 69. 20 μg·ml-1(r = 0. 999 9)for schisantherin,4. 00- 80. 10 μg·ml -1(r = 1. 000 0)for deoxyschizandrin and 3. 42- 68. 30 μg·ml-1(r = 0. 999 9)for schisandrin B. The average recoveries for the five components were all above 98% . Conclusion:The method is simple and accurate with good reproducibility,which can be used for the determination of adenosine,schisandrin,schisantherin,deoxyschizandrin and schisandrin B in Hugan tablets.
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Objective: To establish an RP-HPLC method for simultaneous determining the contents of nine kinds of components in Hugan Agent such as loganin, paeonflorin, scuteuarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid, and to provide the quality guarantee for it. Methods: Agilent Zorbax SB-C18 (150 mm×4.6 mm, 5 μm) column was used. Acetonitrile and 0.1% phosphoric acid water-solution were used as mobile phase of gradient elution, and the volume flow was at 1.0 mL/min; Ultraviolent determination wavelength was 236, 280, and 210 nm; Column temperature was at 30℃, and injection volume was 5 μL. Results: The lowest detection limit in loganin, paeonflorin, scuteuarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid respectively was 12.0, 2.60, 5.75, 9.75, 14.39, 19.06, 14.90, 15.63, and 16.08 ng; The linear range was 583.33-18.229, 916.67-28.65, 541.67-16.93, 416.67-13.02, 500.00-15.63, 458.33-14.32, 625.00-19.53, 458.33-14.32, and 1 000.00-31.25 mg/L, respectively; The average recovery in Hugan Agent was 103.51%, 104.19%, 95.16%, 96.71%, 105.61%, 96.12%, 97.09%, 96.87%, and 105.90%, respectively; The repeatability RSD was 1.39%, 1.41%, 1.87%, 1.91%, 1.79%, 1.45%, 1.32%, 1.71%, and 1.49%, respectively; The stability RSD was 1.36%, 1.22%, 1.87%, 1.91%, 1.93%, 1.56%, 1.39%, 1.78%, and 1.61%, respectively. 6 batche of Hugan Agent contained the average quality of loganin, ursolic acid, paeonflorin, scuteuarin, baicalin, baicalein deoxyschizandrin, schisandrin, and schizandrin respectively was 216.5-222.5, 40.8-42.8, 125.4-136.3, 144.0-147.3, 1640.8-1947.3, 497.5-515.0, 15.0-17.3, 33.6-36.0, and 3.0-3.9 mg/L. Conclusion: The method is believable for determining the concent of loganin, paeonflorin, scuteuarin, baicalin, baicalein, deoxyschizandrin, schisandrin, schizandrin C, and ursolic acid in Hugan Agent with its simplicity, sensibility, repeatability, and better recovery rate.
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Objective:To establish a quantitative analysis of multi-components by single marker ( QAMS) method for the determi-nation of six components ( phillyrin, schizandrol, schisandra ester, deoxyschizandrin, schisandrin B and schisandrin C ) in Liuwei Wuling tablets,and verify the accuracy and feasibility of the method applied in the preparation. Methods:The relative correction factor ( f) of schisandrin B, schisandrin C, schisandra ester, schizandrol, phillyrin and deoxyschizandrin was detected by HPLC at the detec-tion wavelength of 230nm, and the contents of the components in Liuwei Wuling tablets were calculated to achieve the multi evaluation. Meanwhile, the contents of the six components were analyzed by an external standard method and the relative error between the calcula-tion of QAMS method ( Wf ) and the result of the external standard method ( Ws ) was calculated to verify the accuracy of QAMS. The reproducibility of the method was also investigated. Results:The QAMS method was established to determine the contents of 6 compo-nents in Liuwei Wuling tablets, and the components in 10 batches of Liuwei Wuling tablets were determined. The differences between the calculated values and the measured values were small ( RSD < 5%) . Conclusion:The QAMS method is simple, effective and ac-curate in determining the contents of 6 effective components in Liuwei Wuling tablets. The quality evaluation model of QAMS for Liuwei Wuling tablets is validated, which can be used for the quality control of Liuwei Wuling tablets and provide reference for the further study.
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Objective To establish an HPLC method for evaluating deoxyschizandrin in rat plasma; To explore deoxyschizandrin metabolic processes in blood.Methods The blood microdialysis probe was implanted into the right atrial enlargement through right jugular vein to detect the concentration of deoxyschizandrin in blood. Chromatographic conditions for detecting of dialysis fluid were as follows: mobile phase was methanol-acetonitrile- water (40:35:25); flow rate was 1 mL/min; column temperature was 30℃; UV detection wavelength was 254 nm. The data obtained after being calculated by the recovery rate of the probe in vivo were dealt with DAS 2.0 software. Pharmacokinetic parameters were calculated, and the concentration-time related curves were plotted.Results The deoxyschizandrin curve was linear (r=0.999 5) within the range of 2–500μg/mL. The RSD of intra-day pecision and inter-day pecision were both <3%. The stability of the solution was good in 12 hours at roomtemperature. ke was (0.364±0.047)/h; t1/2 was (1.637±0.302)h; Tmax was (0.612±0.194)h; Cmax was (165.92±28.830)μg/mL; AUC0-t was (240.793±25.540)μg?h/mL; AUC0-∞ was (282.710±30.727)μg?h/mL.Conclusion The method is convenient to perform, and the retention time of HPLC is short. This method is suitable for detecting the content of deoxyschizandrin in dialysis fluid, which can also be used to investigate the metabolic processes of deoxyschizandrin in the rat blood.
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Objective: To establish an RP-HPLC method for determining the contents of deoxyschizandrin, schisandrin b, schisandrin c, and ursolic acid in Baogan Pill, and to provide the quality guarantee for Baogan Pill. Methods: Agilent Zorbax SB-C18 (150 mm × 46 mm, 5 μm) column was used. Acetonitrile and 0.1% phosphoric acid water solution was used as mobile phase, the volume flow was at 1.0 mL/min; Ultraviolent wavelength was 210 nm; Column temperature was at 30℃, and injection volume was 10 μL. Results: The lowest detection limits in deoxyschizandrin, schisandrin b, schisandrin c, and ursolic acid respectively were 0.016, 0.019, 0.020, and 0.025 mg/L, the linear ranges respectively were 3.906-250.000, 5.323-340.000, 3.225-205.000, and 5.323-340.000 mg/L. The average recoveries of deoxyschizandrin, schisandrin b, schisandrin c, and ursolic acid in Baogan Pill respectively were 100.79%, 101.09%, 101.26%, and 101.16%; The precision RSD values were 1.76%, 1.69%, 1.80%, and 1.86%, respectively; The repeatability RSD values were 1.77%, 1.66% 1.49% and 1.56%, respectively; The stability RSD values were 1.61%, 1.39%, 1.60%, and 1.56%, respectively. The average amounts of deoxyschizandrin, schisandrin b, schisandrin c, and ursolic acid in the selected samples per pill were 0.444, 1.066, 0.3125, 1.068μg, respectively. Conclusion: The method is believable for determining the contents of deoxyschizandrin, schisandrin b, schisandrin c, and ursolic acid with good simplicity, sensibility, repeatability, and recovery rate.
ABSTRACT
Objective: To establish a UPLC-MS/MS analytical method for the simultaneous analysis of ginsenosides (ginsenosides Rb1, Re, Rg1, Rc, Rd, Rf, Rg3, F2, and notoginsenoside R1) and lignans (gomisin A, schisandrol B, deoxyschizandrin, and schisandrin B) in Yiqi Fumai Injection (freeze-dried) (YFI), and measure the contents of these constituents in YFI. Methods: Quantitative research of 13 components in YFI was done by reversed-phase liquid chromatography on a C18 column using a gradient elution (0.1% formic acid in water and 0.1% formic acid in methanol). A triple quadrupole mass spectrometer operating in positive electrospray ionization mode with multiple reaction monitoring was used. Results: Thirteen components in YFI have good linear relationship, precision, stability, and repeatability according to the requirements of the methodology determination. The recoveries were 98.28%-101.08%. The 13 components in three batches of YFI were determined by UPLC-MS/MS method. Conclusion: The developed UPLC-MS/MS method is simple, sensitive, and accurate, and has good repeatability. The 13 components in YFI could be rapidly and accurately quantified by UPLC-MS/MS, which provides the helpful information for the comprehensive quality evaluation of YFI.