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Objective: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for six active ingredients in Gualoupi injection. Methods: An HPLC method was used with a Waters Sunfire ODS C18column (250 mm×4. 6mm, 2. 5 μm), the mobile phase was methanol(A)-0. 2% glacial acetic acid solution(B) with gradient elution, the flow rate was 1. 0 ml· min-1,the detection wavelength was 254 nm,the column temperature was 30℃ and the injection volume was 20 μl. Using 3, 29-dibenzoyl rarounitriol as a reference, the relative correction factors among karounidiol, vanillic acid, adenosine, quercitrin and cynaro-side were detected by QAMS and their contents were calculated, and the results were compared with those of the external standard method. Results: The differences were not statistically significant between the calculated values of karounidiol, vanillic acid, adeno-sine, quercitrin and cynaroside and those measured by the external standard method (P>0. 05). Conclusion: QAMS can be used for the determination of 6 effective components in Gualoupi injection, and the result is accurate, simple and effective.
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Objective: To determine the contents of five flavonoids in Spirodelae Herba from different growing areas, and evaluate its anti-oxidant activity. Methods: Contents of orientin, vitexin, cynaroside, apigenin-7-glucoside, and luteolin in 17 batches of Spirodelae Herba from eight origins were determined by HPLC with acetonitrile and 0.1% formic acid as mobile phase, and DPPH method was used to compare their anti-oxidant activities, together with cluster analysis and principal component analysis, the quality of Spirodelae Herba was evaluated. Results: The contents of five flavonoids in Spirodelae Herba from different growing areas exist certain differences, which can be divided into three different categories by cluster analysis and principal component analysis. The contents of orientin, vitexin, and cynaroside were significantly higher in samples from Shaanxi and Shanxi provinces than those from other origins, as well as their oxidation clearance rates were relatively high. The correlation analysis showed there was a significant positive correlation between the rate of oxidative elimination and flavonoid content. Conclusion: Spirodelae Herba from Shaanxi and Shanxi provinces has higher contents of flavonoids with strong antioxidant capacity. The analytical method is stable and reliable, which can provide the reference for enhancing the quality control of Spirodelae Herba.
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Objective: To detect the contents of six kinds of flavonoids in Schizonepeta tenuifolia from different areas, so as to provide a basis for their quality control. Methods: Ultra performance liquid chromatography (UPLC) was used to establish the detection scheme of flavonoids content, and clustering analysis was conducted by the results of content detection. Results: The contents of six kinds of flavonoids in S. tenuifolia from different areas had large differences. The linear ranges were: 0.030-0.148 μg for cynaroside, 0.222-1.108 μg for quercetin, 0.357-1.784 μg for hesperidin, 0.058-0.292 μg for luteolin, 0.054-0.269 μg for apigenin, and 0.050-0.247 μg for diosmetin, respectively. The average recovery rate was between 96.32% and 99.97%, RSD < 2.20%; Among this, the Schizonepeta Briq. herb from Anhui was as one class, the herbs from Henan 1 and Guangdong were as one class, and the herbs from Henan 2, Hebei, Yunnan, Sichuan, Hubei, and Gansu were as the other class. Conclusion: The method used for quality control of the multi-index level in this experiment is simple and sensitive; on the other hand, the herbs from Henan 1, Henan 2, Hebei, Yunnan, Sichuan, Hubei, Guangdong and Gansu have a close relationship; They can be used as the same medicinal source.
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Objective: To provide references for quality evaluation of Lonicerea Japonicae Flos (LJF) through exploring correlation between color and contents of active ingredients of LJF in different development periods. Methods: The colorimeter was used to measure the color of LJF in different development periods; HPLC was applied to determine the contents of phenolic acids, flavonoids and iridoids. SPSS 17.0 software was used to analyze the correlation between color and chemical composition of LJF. Results: With the development of flower buds, the contents of phenolic acids, flavonoids compounds achieved the highest in two white stages and the contents of iridoids came to the highest in big white stage and silver flower stage. Therefore, only taking the consideration of the content of active ingredient, the best harvest time of LJF might be the big white stage. There was a significant positive correlation among contents of phenolic acids, flavonoids and brightness (L*) of LJF in different development periods, and there was a very significant negative correlation among contents of phenolic acids, flavonoids, iridoids, and browning index (BI). Conclusion: The color of LJF is closely related to the content of active ingredients, L* and BI can better link appearance and inner quality of LJF.
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OBJECTIVE:To establish the method for pharmacokinetic study of luteolin and cynaroside in rats and to determine pharmacokinetic parameters. METHODS:16 SD rats were randomly divided into luteolin group (sublingual iv,1.34 mg/kg) and cynaroside group(sublingual iv,0.64 mg/kg). 0.5 ml blood were collected before administration and 0,15,30 min and 1,2,3,4,6, 8,12,24,48 h after administration respectively to prepare plasma. UPLC-TQ-MS was adopted to determine plasma concentration, and pharmacokinetic parameters were calculated. A CORTECSTM UPLC? C18(100 mm×2.1 mm,1.6 μm)column was used with mobile phase consisted of acetonitrile-water (containing 0.1% formic acid) at a flow rate of 0.4 ml/min,the column temperature was set at 40 ℃,and quercetin was used as internal standard. RESULTS:The linear range of luteolin and cynaroside were 2.5-500 ng/ml (r=0.998 2) and 10-2 500 ng/ml (r=0.993 5). The lowest quantitation limits were 1 and 2.5 ng/ml,and extraction were 70.75%-87.72% and 75.40%-91.18%(n=6);RSD of inter-day and intra-day were all lower than 10%(n=3). Pharmacokinetic parameters as t1/2 were (1.88 ± 0.32) and (1.57 ± 0.08) h;CL were (0.77 ± 0.18) and (0.06 ± 0.01) L/(h·kg);AUC0-6 h were (189.60±40.04)and(1 093.14±187.36)ng·h/ml;AUC0-∞ were(195.18±38.37)and(1 097.11±188.07)ng·h/ml. CONCLU-SIONS:The method can be used for pharmacokinetic study of luteolin and cynaroside in rats,and the pharmacokinetics of them in rats are in line with two-compartment model.
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The present study was designed to establish and optimize a new method for extracting chlorogenic acid and cynaroside from Lonicera japonica Thunb. through orthogonal experimental designl. A new ultrahigh pressure extraction (UPE) technology was applied to extract chlorogenic acid and cynaroside from L. japonica. The influential factors, including solvent type, ethanol concentration, extraction pressure, time, and temperature, and the solid/liquid ratio, have been studied to optimize the extraction process. The optimal conditions for the UPE were developed by quantitative analysis of the extraction products by HPLC-DAD in comparison with standard samples. In addition, the microstructures of the medicinal materials before and after extraction were studied by scanning electron microscopy (SEM). Furthermore, the extraction efficiency of different extraction methods and the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of the extracts were investigated. The optimal conditions for extracting chlorogenic acid and cynaroside were as follows: ethanol concentration, 60%; extraction pressure, 400 MPa; extraction time, 2 min; extraction temperature, 30 °C; and the solid/liquid ratio, 1 : 50. Under these conditions, the yields of chlorogenic acid and cynaroside were raised to 4.863% and 0.080%, respectively. Compared with other extraction methods, such as heat reflux extraction (HRE), ultrasonic extraction (UE), and Sohxlet extraction (SE), the UPE method showed several advantages, including higher extraction yield, shorter extraction time, lower energy consumption, and higher purity of the extracts. This study could help better utilize L. japonica flower buds as a readily accessible source of natural antioxidants in food and pharmaceutical industries.
Subject(s)
Analytic Sample Preparation Methods , Methods , Antioxidants , Chlorogenic Acid , Chromatography, High Pressure Liquid , Flowers , Chemistry , Glucosides , Lonicera , Chemistry , Luteolin , Plant Extracts , PressureABSTRACT
In this study, we investigated whether cynaroside, cynarin and linarin derived from Chrysanthemum indicum L. affect the secretion, production and gene expression of MUC5AC mucin in airway epithelial cells. Confluent NCI-H292 cells were pretreated with cynaroside, cynarin or linarin for 30 min and then stimulated with PMA (phorbol 12-myristate 13-acetate) for 24 h. The MUC5AC mucin gene expression, mucin protein production and secretion were measured by RT-PCR and ELISA, respectively. Effect of linarin on EGF (epidermal growth factor) - or TNF-alpha (tumor necrosis factor-alpha)-induced MUC5AC mucin gene expression and mucin protein production was also examined. The results were as follows: (1) Cynaroside and cynarin did not significantly affect PMA-induced MUC5AC mucin secretion from NCI-H292 cells. However, linarin decreased MUC5AC mucin secretion; (2) Cynaroside did not affect PMA-induced MUC5AC mucin production and gene expresion from NCI-H292 cells. However, cynarin and linarin inhibited the production and gene expression of MUC5AC mucin; (3) Linarin also inhibited the production and gene expression of MUC5AC mucin induced by EGF- or TNF-alpha from NCI-H292 cells. These results suggest that linarin can regulate the gene expression, production and secretion of mucin, by directly acting on airway epithelial cells.
Subject(s)
Chrysanthemum , Enzyme-Linked Immunosorbent Assay , Epidermal Growth Factor , Epithelial Cells , Gene Expression , Mucins , Necrosis , Tumor Necrosis Factor-alphaABSTRACT
Objective: To establish a UPLC-MS/MS method for simultaneously determining ten components (chromogenic acid, liquiritin apioside, calycosin-7-glucoside, cynaroside, liquiritin, ferulic acid, isoliquiritoside, harpagoside, liquiritigenin, and cinnamic acid) in Tongsaimai Pellets. Methods: A method for the simultaneous determination of the ten components was established by UPLC-MS/MS. The analysis was performed on a Waters Acquity BEH C18 column (100 mm × 2.1 mm, 1.7 μm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL/min. The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source combined with multiple reaction monitoring (MRM) mode. Results: The linear relationships between the concentration and peak areas of the ten components were chromogenic acid Y = 294.09 X + 17624; liquiritin apioside Y = 19.296 X + 748.42; calycosin-7-glucoside Y = 670.8 X + 11121; cynaroside Y = 490.45 X + 2938.3; liquiritin Y = 33.489 X + 555; ferulic acid Y = 127.76 X + 1343.5; isoliquiritoside Y = 57.87 X + 804.81; harpagoside Y = 13.125 X-20.365; liquiritigenin Y = 29.057 X + 367.71; cinnamic acid Y = 72.867 X + 1301.5 (r > 0.9990); The precisions, repeatabilities, and stabilities of the method were satisfactory, and the average recoveries were between 96.00% and 104.67% with the relative standard deviations no more than 3%. Conclusion: The established method is specific, rapid, and accurate for the determination of the ten effective components in Tongsaimai Pellets.
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Monoamine oxidase inhibitors (MAOI) have been widely used as antidepressants. Recently, there has been renewed interest in MAO inhibitors. The activity-guided fractionation of extracts from Angelica keiskei Koidzumi (A. keiskei K.) led to the isolation of two prenylated chalcones, xanthoangelol and 4-hydroxyderricin and a flavonoid, cynaroside. These three isolated compounds are the major active ingredients of A. keiskei K. to inhibit the MAOs and DBH activities. Xanthoangelol is a nonselective MAO inhibitor, and a potent dopamine beta-hydroxylase (DBH) inhibitor. IC50 values of xanthoangelol to MAO-A and MAO-B were calculated to be 43.4 microM, and 43.9 microM. These values were very similar to iproniazid, which is a nonselective MAO inhibitor used as a drug against depression. The IC50 values of iproniazid were 37 microM, and 42.5 microM in our parallel examination. Moreover, IC50 value of xanthoangelol to DBH was calculated 0.52 microM. 4-Hydroxyderricin is a potent selective MAO-B inhibitor and also mildly inhibits DBH activity. The IC50 value of 4-hydroxyderricin to MAO-B was calculated to be 3.43 microM and this value was higher than that of deprenyl (0.046 microM) used as a positive control for selective MAO-B inhibitor in our test. Cynaroside is a most potent DBH inhibitor. The IC50 value of cynaroside to DBH was calculated at 0.0410 microM. Results of this study suggest that the two prenylated chalcones, xanthoangelol and 4-hydroxyderricin isolated from A. keiskei K., are expected for potent candidates for development of combined antidepressant drug. A. keiskei K. will be an excellent new bio-functional food material that has the combined antidepressant effect.