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Objective To investigate the extraction methods for active components from oral ulcer film and optimize the determination methods of active components dexamethasone sodium phosphate and metronazole. Methods Different extraction solvents(methanol, water and 70% methanol aqueous) were applied to extract the active components dexamethasone sodium phosphate and metronazole from oral ulcer film, which contents were quantified by a HPLC method. Results the extraction solvent water had the best efficacy and more simpler compared to the other two solvents. Clotriazole showed a good linear relationship within 5.014 5-200.5800 μg/ml (r=0.999 8), and the average extraction recovery was (104.23±0.63)%, and for dexamethasone sodium phosphate, a good linear relationship was obtained in the range of 0.482-16.328 μg/ml (r=0.9999), and the average extraction recovery was (103.97±1.02)%. Conclusion The water extraction method established in this study was simple and efficient, which showed features of simplicity, accuracy and repeatable.
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Objectives To investigate the effects of cell aging on the disorders relating to gastric mucosa aging.Methods A treatment of 200 μmol/L H2O2 was used to induce senescence of human gastric epithelial cell line GES-1,and the cell growth curve was monitored.Senescence secretory phenotypes were observed by detecting the protein level of p53 and p16INK4a with senescence-associated β-galactosidase(SA-β gal)staining and Western blot testing.The mRNA levels of senescence-associated secretory phenotype(SASP)factors in human gastric epithelial GES-1 cell including IL-1β,IL-6,IL-8,TGF-β、IFN-γ,and VEGF-A were detected by RT PCR.The mRNA expression levels of IL-1β,IL-6,IL-8,TGF-β,IFN-γ,and VEGF in the conditioned medium were detected by ELISA analysis.Results The 200 μmol/L H2O2-induced GES-1 cells stopped proliferating after 3 days of treatment,and cells enlarged and flattened at 10 days.The increased SA-β-gal staining(P＜0.001) and the increased expression levels of p53 and p16INK4a proteins indicated the success of establishing the aging model of GES-1.The mRNA levels of IL 1β,IL6,IL8,TGF-β,and IFNγ were higher(t=2.94,3.38,3.15,3.64,2.97;P=0.015,0.000,0.000,0.000,0.000)and the mRNA level of VEGF-A was lower(t=2.31,P =0.20) in senescent GES-1 cells than in the control group.In the conditioned medium of senescent GES-1 cells,the levels of IL-1β,IL6,IL8,TGF-β1,and IFNγ were higher in the H2O2-induced group [(3.12±0.21)μg/L,(4.26±0.15)μg/L,(3.37±0.14)μg/L,(5.34±0.19)μg/L,and(2.90±0.47)μg/L]than in the negative control group[(0.24±0.04,0.04±0.07,0.52±0.02,1.05±0.10,0.52±0.02,respectively,P＜0.001)],while the level of VEGF was lower in the H2O2-induced group than in the negative control group(0.21±0.03)μg/L vs (0.59±0.07)μg/L(P＜0.05).Conclusions The changes in senescence-associated secretory phenotype factors of the aging human gastric epithelial cells induced by oxidative stress may promote chronic gastritis and gastric cancer.
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Objective To establish the quality standard of Santeng oral solution.Method Sargentodoxa Caulis and Spatholobi Caulis were identified by thin layer chromatography.Chlorogenic acid was assayed by high performance liquid chromatography.The chromatographic column is Agilent Zorbax SB C18 (4.6 mm×250 mm, 5 μm) with a stable temperature of 35 ℃.The mobile phase in isocratic elution consists of acetonitrile and 0.1% folic acid aqueous solution with a preliminary volume ratio of 9∶91.The flow rate is 1.0 ml/min with an injection volume of 20 μl.Results Thin layer chromatography showed distinct spots of Sargentodoxa Caulis and Spatholobi Caulis with a great specificity.A regression formula Y=60.14X-6.37(r>0.999 9) was obtained with a good linearity in concentration range of 2.70~202.50 μg/ml.Conclusion A simple, stable and repeatable method was established for the quality control of Santeng oral solution.
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Objective To establish a quality control standard for Qingre Baidu granules .Methods Isatidis Radix ,Fruc-tus Forsythiae ,Herba Violae ,and Glycyrrhizae were identified by TLC ,and the concentration of chlorogenic acid was deter-minedbyHPLC.ThismethodwasemployedonanAgilentZORBAXSB-C18column(4.6mm×250mm,5μm)at30℃ witha mobile phase of acetonitrile (A) and 0 .2% formic acid (B) using the gradient elution program shown as follows :0-12 min , 11%-12% A run at the flow rate of 1 .0 ml/min .The injection volume was 20 μl and the detection wavelength was 327 nm . Results Characteristic spots could be detected by TLC and the specificity of the method was satisfactory .As for chlorogenic acid ,the equation of linear regression of chlorogenic acid was Y=60 .239 4X+9 .096 3 (r=0 .999 9) with the linear range of 6.19-396 .00 μg/ml .The average recovery was 99 .66% (RSD=2 .82% ) .Conclusion The established method is simple ,reli-able ,reproducible ,and can be used for the quantitative determination and quality control of Qingre Baidu granules .
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Objective To establish a high performance liquid chromatography method for the determination of matrine in Lefu oral liquid .Methods The HPLC method was performed on a Diamonsil Platisil NH2 column (4 .6 mm × 250 mm ,5μm) with the mobile phase of acetonitrile-isopropyl alcohol-3% phosphoric acid solution (84 ∶ 4 ∶ 12 ) . The flow rate was 1 .2 ml/min .The sample injection volume was 5 μl .The detective wavelength was 205 nm .Results The calibration curve of matrine showed good linear response ranged from 54 .50 to 872 .00 μg/ml with r=0 .999 1 .The average recovery of spiked samples for matrine was 99 .82% while the relative standard deviation for repetitions was 1 .12% .Conclusion The method was simple ,reliable and repeatable ,which could be used for the quantitative determination of matrine of Lefu oral liquid .
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Objective To establish quality control of Xinnaoling granules .Methods Thin layer chromatography (TLC) methods were used to identify Rhizoma Corydalis ,Radix Angelicae dahuricae ,Cortex Magnoliae officinalis ,Radix Glycyrrhi-zae ,Rhizoma Ligusticum ,Ligusticum wallichii ,Schisandra Chinensis ,Dendranthema morifolium .The concentration of salvi-anolic acid B was determined by high performance liquid chromatography (HPLC) .The method employed a column of Agilent Eclipse XDB-C18 (4 .6 mm × 250 mm ,5 μm) with a mobile phase of 0 .5% formic acid (A)-acetonitrile (B) at a temperature 25℃ .The gradient elution program was as follow :0~5 min 24% B ,5~6 min 24% ~19% B ,6~25 min 19~20% B .The flow rate was 1 .0 ml/min ,and the injection volume was 10μl and the detection wavelength was 286 nm .Results The spots in TLC plates were clear and specific .As for salvianolic acid B ,the linear range was 20-1 280 μg/ml and the equation of linear regres-sion of salvianolic acid B was Y=13 .304 X -117 .50 (r=0 .999 9 ,n=7) .The average recovery rate was 96 .17% (RSD=1 . 10% ) .Conclusion The method was proved to be simple ,reliable ,reproducible ,and could be used in the quality control of Xinnaoling g ranules .
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Objective To study the fingerprint of Polygonum cuspidatum from various regions by HPLC and provide technical support for the identification.Methods HPLC conditions :the chromatographic column was Agilent ZORBAX SB-C18 (4.6 mm × 250 mm,5 μm);the mobile phase was 0.1% formic acid in water and acetonitrile (A-B);gradient elution ;the de-tection wavelength was 306 nm ;the flow rate was 1 ml/min;the column temperature was 30 ℃ ;injection volume was 20 μl. Results The HPLC gradient conditions could effectively separate polydatin,resveratrol,emodin and emodin ether,etc.Con-clusion This method is stable,effective and durable,which is instructive to the herbs data identification of Polygonum cuspi-datum.
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Objective To establish the quality standard for Quban oral liquid .Method Thin layer chromatography (TLC)was used for quality identification of Codonopsis pilosula ,Astragalus membranaceus ,Paeonia veitchii ,Paeonia lacti‐flora,AtractylodesmacrocephalaandAngelicasinensis.ThepaeoniflorinwasdeterminedbyRP‐HPLC.Result Clearspots were obtained with good separation in TLC identification which was highly specific .The paeoniflorin area and content showed a good linear relationship at the range of 18 .20‐2 330 .00 μg/ml (r= 0 .999 9) .The average recovery was 99.72% (RSD=1.25% ) .Conclusion This method is simple ,rapid ,accurate ,and which can be used for the quality control of Quban oral liquid .
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OBJECTIVE:To study the inhibitory mechanism of timosaponin B-Ⅱ(TB-Ⅱ) on the proliferation and migration of human lung cancer A549 cells. METHODS:A549 cells were treated with TB-Ⅱ [0(blank control),1,10 and 100 μg/ml] for 48 h,and total RNA and total protein were extracted respectively. Real time fluorescence quantitative-PCR and Western blot were used to detect mRNA and protein levels of IL-18. IL-18 in A549 cells was silenced by transfection;the expression of IL-18 mRNA and protein were compared among untransfection group,negative control group and transfection group;and then human lung can-cer A549 cells with silenced gene were treated with 10 μg/ml TB-Ⅱ for 24,48 and 72 h. The activity of cell proliferation was de-tected with CCK-8,and the change of cell migration ability was observed by streak method. RESULTS:Compared with blank con-trol,the expression of IL-18 mRNA and protein in A549 cells all increased after treated with TB-Ⅱ(P<0.05 or P<0.01),and were positively correlated with concentration. Compared with untransfection group,the expression of IL-18 mRNA and protein de-creased in transfection group(P<0.01). Compared with untransfected cell treated with TB-Ⅱ,the viability and migration ability of A549 cells with transfection gene increased after treated with TB-Ⅱ for 72 h(P<0.01). CONCLUSIONS:TB-Ⅱ can inhibit the proliferation and migration of A549 cells by up-regulating IL-18 gene expression.
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Objective To establish quality standards forAnshu Liquid.Methods Phellodendri Chinensis Cortex and Kochiae Fructus were qualitatively identified by TLC method. The contents of berberine hydrochloride and phellodendrine hydrochloride in the preparation were determined by HPLC method. The chromatographic column was ZORBAX SB-C18 (4.6 mm×250 mm, 5μm); the mobile phase was acetonitrile-0.1% phosphoric acid (0.5% diethylamine) gradient elution; the flow rate was 1.0 mL/min; the detection wavelength was 284 nm.Results Berberine hydrochloride and phellodendrine hydrochloride in Phellodendri Chinensis Cortex and saponin Ic in Kochiae Fructus were detected by TLC method. The linear relationship of berberine hydrochloride in the preparation was Y=23.109X?30.548,r2=0.999 8, RSD=2.97%, showing that the linear range of berberine hydrochloride was 0.050 5– 2.525 0 μg. The linear relationship of phellodendrine hydrochloride in the preparation wasY=10.038X?2.446 6, r2=0.999 9, RSD=1.24%, showing that the linear range of phellodendrine hydrochloride was 0.050 0–2.500 0 μg. Conclusion The method is accurate, rapid, stable and reliable, with good reproducibility, which can be used for the quality control and evaluation ofAnshu Liquid.
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Objective To establish a quality standard forZishen Tongguan Capsules.Methods Anemarrhneae Rhizoma and Phellodendri Chinensis Cortex were qualitatively identified by TLC method. The contents of neomangferin, phellodendrine hydrochloride, mangiferin and berberine hydrochloride inZishen Tongguan Capsules were quantitatively determined by HPLC method.Results The spots of qualitative identification method were clear without interference. Neomangferin in Anemarrhneae Rhizoma and phellodendrinehy drochloride in Phellodendri Chinensis Cortex were identified. Neomangferin, phellodendrinehy drochloride, mangiferin and berberine hydrochloride showed good linearity in the range of 0.025 25–2.02 μg (r=0.999 7), 0.025 5–2.04 μg (r=0.999 7), 0.026–2.08 μg (r=0.999 7), and 0.025 5–2.04 μg (r=0.999 6) respectively. The average recoveries of neomangferin, phellodendrinehy drochloride, mangiferin and berberine hydrochloride were 99.99%, 101.06%, 103.05%, and 100.55%, respectively. The RSD were 2.69%, 5.62%, 2.49%, and 2.06%, respectively.Conclusion The method is accurate and rapid, with good stability, reliability and reproducibility, which can be used for the quality control and evaluation of the preparation of Zishen Tongguan Capsules.
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OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.
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Objective To establish the quality standards for compound Shatagan Oral Liquid. Methods Chuanxiong Rhizoma was identified by TLC. The content of ferulic acid was determined by HPLC. Separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5μm) with a mobile phase consisting of methanol-1% acetic acid solution in gradient elution (0-5 min, 35% methanol;5-8 min, 35%→23% methanol;8-22 min, 23% methanol) at 30℃;The flow rate was 1.0 mL/min;The injection volume was 5μL;The detection wavelength was 322 nm.Results Ferulic acid showed a good linear relationship in the range of 0.039 4-0.630 0μg (r=0.999 7,n=7). The average recovery was 98.22% and RSD was 2.62% (n=6).Conclusion The method is reliable, sensitive and with repeatability, which can be used as the quality control method for compound Shatagan Oral Liquid.
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OBJECTIVE:To establish the quality standard for Xianrengu oral liquid. METHODS:Thin layer chromatography (TLC) was used for qualitative identification of Epimedii Folium,Astragali Radix,lycium barbarum,Atractylodes macrocephala and Panax ginseng in Xianrengu oral liquid;RP-HPLC was preformed on the column of Dikma Diamonsil C18 with the mobile phase of acetonitrile-0.1% phosphoric acid (30∶70,V/V)at the flow rate of 1.0 ml/min,the detection wavelength was 270 nm,the temperature was 30℃and the volume was 10μl. RESULTS:TLC identification had good separation,clear spots and high specifica-tion. The linear range of icariin was 7.97-510.00 μg/ml(r=0.999 3);RSDs of precision,stability and reproducibility tests were no more than 0.83%;and the average recovery was 97.69%(RSD=1.30%,n=6). CONCLUSIONS:This method is simple,accu-rate and simple,which can be used for the quality control of Xianrengu oral liquid.