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An ultra-high performance liquid chromatography method for the determination of 8 constituents in Qingzao Jiufei Decoction was established and the basis of related chemical substances with antioxidant activity in Qingzao Jiufei Decoction was explored. The separation was performed on a Waters Cortecs RP Shield C18 (150 mm × 2.1 mm, 1.6 μm) using UHPLC-DAD as the mobile phase was water (containing 0.1% phosphoric acid) – acetonitrile with flow rate of 0.30 mL·min-1 by gradient elution ① determining 5 constituents (amygdalin, liquiritin, liquiritin apioside, rutin and isoquercitrin) at the wavelength of 210 nm, 237 nm and 358 nm. Under gradient elution ②, 3 constituents (glycyrrhizin, glycyrrhizic acid and sesamin) were determined at the wavelength of 210 nm and 265 nm. The IC50 of 10 batches of Qingzao Jiufei Decoction scavenging 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS+) free radicals obtained through test and Probit model was analyzed for correlation with the contents of 8 constituents. The established methods had a good linear relationship (r > 0.999), good repeatability and stability. The recovery rate was between 82.8% and 112.4%. In a series of concentration range, the higher the concentration of Qingzao Jiufei Decoction, the stronger the free radical scavenging effect. There was a significant correlation between the content of rutin and glycyrrhizic acid and the IC50 of scavenging free radicals. The content determination methods established in this experiment provide a basis for a reasonable and scientific evaluation of the quality of Qingzao Jiufei Decoction. Qingzao Jiufei Decoction has antioxidant activity, which is significantly positively correlated with the content of rutin and glycyrrhizic acid.
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A UHPLC method for the simultaneous determination of multiple constituents in QingJinHuaTan Decoction was established. The separation was performed on a Waters cortecs T3 column (150 mm×2.1 mm, 1.6 μm); the mobile phase was acetonitrile-water (containing 0.04% phosphoric acid) with gradient elution at a flow rate of 0.30 mL·min-1, the column temperature at 25 ℃ and the wavelengths at 238 nm and 280 nm. The results showed that all peaks were well separated and all components had a good linear relationship in the investigative range, (r > 0.999). The repeatability and stability were good and the recovery was between 92.5%-104.7%. The method is simple, accurate and reliable and provides a basis for quality control of QingJinHuaTan Decoction and for further development of methods for its standardization.
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Objective:To analyze the known mechanism of toxicology and predict the unknown toxicity in Asari Radix et Rhizoma sinensis by establishing the network relationship of compound, protein, gene and toxicant reaction. Method:After comparing the Asari Radix et Rhizoma candidate compounds obtained from the traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) database and the toxicological information obtained from the Comparative Toxicogenomics Database(CTD) database, we screened out 13 toxic components from Asari Radix et Rhizoma. And use the Pharm Mapper Server website to find the detailed information of target proteins of the 13 components. The network structure of these 13 chemical components and their corresponding target proteins were drawn by using Cytospace software, and several target proteins with the highest degree of association were found. ClueGO+CluePedia plug-in of Cytospace software was applied in gene ontology(GO) enrichment analysis of genes and kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis, so as to determine the pathways through which toxic substances in Asari Radix et Rhizoma might be harmful to human body. Result:The toxic substances in Asari Radix et Rhizoma may induce tumor and cancer formation through p53 signaling pathway, interleukin(IL)-17 signaling pathway, nuclear factor(NF)-kappa B signaling pathway, tumor necrosis factor(TNF)-signaling pathway. Asari Radix et Rhizoma could inhibit the central nervous system by regulating apoptosis pathways and neurons, and may also cause other autoimmune diseases by IL-17, TNF-α pathway and apoptosis regulation. Conclusion:This study preliminarily explores related mechanisms of toxicity of Asari Radix et Rhizoma,this method can be used to predict toxicity and explain toxicity mechanism of traditional Chinese medicine.
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Objective: To explore the mechanism of renal toxicity of Tripterygii Radix et Rhizoma by establishing the active component-target, protein interaction, biological function and pathway network corresponding to the target, and using molecular docking technology. Method: The traditional Chinese medicine(TCM) systems pharmacology database(TCMSP) and the comparative toxicogenomics database (CTD) were used to screen The toxic candidate compounds.In PubChem database, convert all candidate compounds into standard Canonical SMILES format, SMILES format file import SwissTargetPrediction platform, target prediction, will be the target of the corresponding compounds in TCMSP supplement with uniprot converts protein antipodal gene name, and from the human genome database (GeneCards) seek to compare the renal related gene protein,overlapping proteins were screened as potential renal toxicity targets of Tripterygii Radix et Rhizoma.Cytoscape software was used to construct the candidate components-target network of Tripterygii Radix et Rhizoma.Cytoscape software was combined with String database to draw the protein interaction network, DAVID platform was used to analyze the biological function of the target and the pathways involved, and Glide software was used to verify the combination of the key protein and the candidate components of tripterygiumwildiitoxicity. Result: The screening of 30 kinds of candidates for toxic ingredients of Tripterygii Radix et Rhizoma, involving 209 renal toxicity targets, network analysis results showed that Tripterygii Radix et Rhizoma by amino acid metabolism,phospholipid metabolism, catecholamine metabolism, inhibiting renal organic anion transporter Oatl, Oat2, Oat3 function, and inducing apoptosis, and participate in the mitogen-activated protein kinase(MAPK) signaling pathways, JAK-STAT signaling pathway,vascular endothelial growth factor(VEGF)signaling pathways,Toll-like receptor signaling pathway,ERBB signaling pathway, FcεRI signaling pathway, peroxisome proliferators-activated receptors(PPAR) signaling pathway such as toxic to the kidneys. Conclusion: The mechanism of kidney toxicity of Tripterygii Radix et Rhizoma was explored by using the characteristics of multi-component, multi-target and multi-pathway of TCM, which provided new ideas and methods for further research on the mechanism of kidney toxicity of Tripterygii Radix et Rhizoma.
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Objective:The mechanism of action of cardiac toxicity of Radix Aconiti Agrestis was explored by establishing the active components-targets network of Radix Aconiti Agrestis, protein interaction network, the biological function and pathway network of targets, and using molecular docking technology. Methods:The Traditional Chinese Medicine Systems Pharmacology(TCMSP) database and the Comparative Toxicogenomics Database(CTD) were used to filtrate the toxic candidates of Radix Aconiti Agrestis. Predicting the functional targets of toxic candidates of Radix Aconiti Agrestis by PharmMapper and compared with the cardiac related gene proteins found in the human gene database (GeneCards), and the overlapping proteins were selected as potential cardiac toxicity targets of Radix Aconiti Agrestis. The Cytoscape software was used to construct the network between toxic candidate components and targets. The protein interaction network was mapped by the String database combined with Cytoscape software. The biological functions of the targets and the involved pathways were analyzed with the DAVID platform.The binding of the key proteins with certain toxic candidate components of Radix Aconiti Agrestis was verified by Discover Studio software finally. Results:There were six candidates for toxic ingredients, which involving 27 cardiac toxicity targets. Network analysis results show that the targets were mainly by participating in the heart of phosphorus metabolism, regulation and other related phosphorus metabolism and regulation of phosphorylation and FKBP1A,TGF4-β2, INSR targets to have an important impact on the metabolism,development and form of the heart,and further to have cardiac toxicity. Conclusion:Based on the characteristics of the multi-component, multi-target and multi-pathway of traditional Chinese medicine, the mechanism of cardiac toxicity of Radix Aconiti Agrestis was explored and its possible toxicity was predicted, which provided a new idea and method for further research on the mechanism of cardiac toxicity of Radix Aconiti Agrestis.
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A quantitative analytical method for multi-components with a single-marker (QAMS) was established for simultaneous determination of neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction. The separation was performed on a Waters CORTECS T3 column (2.1 mm × 100 mm, 2.7 μm), with the mobile phase consisting of 0.05% phosphate acid solution-acetonitrile for gradient elution. The column temperature was 30 ℃, and flow rate was 0.5 mL·min-1. Using chlorogenic acid as an internal reference, the relative correlation factors of neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid were calculated following UHPLC, as 0.928 0, 0.546 2, 1.099 8, 0.872 1, 1.086 8, 0.739 2, 1.056 6, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method. There was no significant difference in assay results between QAMS and the external standard method. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the content such as neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction.
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For qualitative and quantitative analysis of related substances in clotrimazole cream, HPLC-Q-TOF spectrometer was used to analyze the fragmentation pathways and identify structures of the related substances. Five related substances named by BP (2018) were identified as impurity A ((2-chlorophenyl)-diphenylmethanol), impurity B (para-clotrimazole isomer), impurity E (2-chlorobenzophenone), impurity F (1-tritylimidazole) and impurity 4 (9-(2-chlorophenyl)-fluorene), respectively, by using impurity references matching and comparison with the literature data. Four related substances were detected in clotrimazole cream except impurity E, and 9-(2-chlorophenyl)-fluorene is the first identified impurity in this preparation. To establish an HPLC method for determination of the related substances in Clotrimazole Cream, the Agilent Poroshell Bonuns RP column was used (100 mm×4.6 mm, 2.7 μm) with UV detection at 215 nm. The mobile phase was acetonitrile-10 mmol·L-1 dipotassium phosphate buffer (adjusted with phosphoric acid to pH of 5.80) with a flow rate of 1.0 mL·min-1. Gradient elution was used. The column temperature was maintained at 40℃. A good linear behavior was achieved between component's concentrations and peak area for impurity A, B, E, F within the range of 0.20-10.02 μg·mL-1, 0.20-10.00 μg·mL-1, 0.20-10.10 μg·mL-1, 0.10-5.01 μg·mL-1 with the correlation coefficients were 0.999 7, 1.000 0, 1.000 0, 0.999 9, respectively. The average recoveries were 94.3%, 95.0%, 100.0%, 99.6% with RSDs were 2.8%, 2.2%, 1.1%, 2.7%, respectively (n=9). LOQ were 200.4, 200.0, 202.0, 100.2 ng·mL-1, respectively. LOD were 57.25, 57.14, 57.71, 28.63 ng·mL-1, respectively. The developed method was simple, rapid, accurate and effective for testing related substances in clotrimazole cream to control its quality, ensuring the safety of clinical medication.
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An HPLC method was established for the simultaneous determination of saikosaponin a, b2, c, d, e, f of Bupleurum chinense DC. in order to study the content difference of saikosaponins in different producing areas, different harvest time and different processed products of Bupleurum chinense DC. The Agela Venusil MP C18 (4.6 mm×250 mm, 5 μm) column was used with a gradient elution of acetonitrile-water at the wavelength of 210 and 254 nm with the flow rate of 1.0 mL·min-1 and the column temperature at 30℃. Based on the content of six kinds of saikosaponins, the differences of saikosaponins in four producing areas, eight harvest periods and 11 processing methods of Bupleurum chinense DC. were systematically studied. The results showed that the content of saikosaponins in Bupleurum chinense DC. was higher in May and August of Liaoning, Shaanxi and Gansu, but only in August from Shanxi in the four producing areas. The content of saikosaponins in 11 processed products was as follows:raw product > bran-stir-fried product > stir-fried product > wine-moistened product > turtle blood-stir-fried product > bran-wine-stir-fried product > wine-stir-fried product > vinegar-moistened product > turtle blood-wine-stir-fried product > vinegar-stir-fried product > honey-stir-fried product > honeymoistened product.
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The study is aimed to develop a method in evaluation of the bioactive consistency of cardiotonic pill (CP). HepG2 cell line was employed as a biological detector. After treated with CP for 24 h, gene chip and qRT-PCR were used to select mRNAs that can represent the bioactivity of CP. Then similarity between different batches of CP were calculated based on expression levels of marker genes to evaluate the bioactive consistency of CP. Marker genes were selected according to the criteria as follows:① fold change 1.5; ② potential relevance to curative effects; ③ extensive involvement in the cellular functions and clustering analysis categories; ④ dose-dependent effect. A total of 10 genes were selected as bioactive markers of CP. Angular cosine was calculated to evaluate the similarity between two samples. The method was validated using intra-day precision and inter-day precision. Using angular cosine similarity, the intra-day and inter-day precision were 0.4% and 0.6%, respectively. The similarities of 6 batches of CDPs ranged from 0.992 to 0.999, and 1 batch of Compound Danshen Tablet was 0.534. The established method is specific and accurate, and provides comprehensive and objective evaluation of bioactive quality of CDPs. It can also benefit the bioactive consistency evaluation of other compounds in traditional Chinese medicines.
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Two regression models, based on panel data over the period of 2000-2011, are built and used to analyze what factors determine China's exports of primary and semi-finished products of traditional Chinese medicine to ASEAN. The results indicate that, China GDP, the ratio of ASEAN to China GDP per capita, average export price, the ratio of state-owned assets to total assets, have a significant positive influence on the export volumes of primary products of Chinese medicine. At the same time, RMB appreciation, the ratio of three kinds of foreign-invested assets to total assets, China-ASEAN Early Harvest Program, ASEAN-China Free Trade Area have a significant negative influence. In respect of the export volumes of semi-finished products of Chinese medicine, the significant influential factors are ASEAN GDP and the ratio of ASEAN to China GDP per capita. The former is positive and the latter is negative. In order to optimize the commodity composition of experts, it is needed to increase export volumes of both primary and semi-finished products of Chinese medicine. According to the analysis above, some proposals are put forward, such as, improving the performance of foreign capital, playing an exemplary and leading role in technological innovation by state-owned enterprises, taking advantage of bargaining power of suppliers, increasing outward foreign direct investment.
Subject(s)
China , Commerce , Drugs, Chinese Herbal , Chemistry , Economics , Europe, Eastern , Medicine, Chinese Traditional , Economics , Reference Standards , Quality ControlABSTRACT
An HPLC method has been developed to determine polydatin in giant knotweed rhizome. In order to systematically validate the method, specificity, precision, linearity of reference solution and test solution, repeatability, reproducibility, accuracy, stability and robustness were measured. In the robustness test, a one-variable-at-a-time procedure was applied to evaluate the influence of slight variations in method factors, including the flow rate, the column temperature, the extraction time, and etc., on the assay result of polydatin. No significant differences were found when the process parameters changed during the experimental domain. And system suitability test limits were defined based on the robustness test. Results showed that the developed method was accurate, reproducible and robust.
Subject(s)
Chromatography, High Pressure Liquid , Methods , Drug Stability , Fallopia japonica , Chemistry , Glucosides , Plants, Medicinal , Chemistry , Reproducibility of Results , Rhizome , Chemistry , Sensitivity and Specificity , StilbenesABSTRACT
<p><b>OBJECTIVE</b>To look for the active fraction of ethanol extract of Genkwa Flos (EGF) induced hepatotoxicity and develop an UPLC fingerprint of the active fraction.</p><p><b>METHOD</b>Target fraction of EGF induced hepatotoxicity was guided by the serum biochemical and histopathology methods. The UPLC method was applied to establish the chromatographic fingerprint. The separation was achieved on a BEH C18 column (2.1 mm x 50 mm, 1.7 microm) with a mobile phase consisting of acetonitrile and water containing 0.05% phosphate acid running gradient elution. The detection was carried out at 210 nm and the analysis was finished within 10 min.</p><p><b>RESULT</b>The chloroform phase of EGF could be responsible for the hepatotoxicity of this herb. The common mode of the UPLC fingerprint was set up under the established condition. There were 17 common peaks in fourteen batches of herbs, eight of which were identified, and the similar degrees of the fourteen batches to the common mode were between 0.890-0.999.</p><p><b>CONCLUSION</b>It is easy to locate the chloroform extraction of EGF with hepatotoxicity. And the UPLC fingerprint was developed for the above fraction, which could provide valuable references for safe and effective clinical use of EGF.</p>
Subject(s)
Animals , Humans , Male , Rats , Asteraceae , Chemistry , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Toxicity , Flowers , Chemistry , Liver , Rats, WistarABSTRACT
<p><b>OBJECTIVE</b>To develop a LC-MS method for the determination of senkyunolide I (SI) in rat plasma, in order to observe whether there is significant change in the pharmacokinetics parameters of complex prescriptions of Huoluoxiaolingdan (HLXL) and single herbal extracts from Ligusticum chuanxiong Hort. in rats, and assess the effect of other components in HLXL on the pharmacokinetics of SI.</p><p><b>METHOD</b>Twelve male Sprague-Dawley (SD) rats were randomly divided into two groups, and orally administered with extract from HLXL and L. chuanxiong (both equal to SI 4.53 mg x kg(-1)). Their blood was collected at different time points for LC-MS, in order to detect the plasma concentration of SI. The pharmacokinetic parameters of SI were calculated by DAS 2.0 software. SPSS 16.0 software was used for independent-sample T-test and Nonparametric T-test.</p><p><b>RESULT</b>A linear relationship of SI ranged from 6.750 to 675.0 microg x L(-1), and with the lowest limit of detection being 6.750 microg L(-1). Both of the plasma concentration-time curves of SI were fitted with the two-compartment model for extract of HLXL and L. chuanxiong. The detected AUC and Cmax of SI showed significant difference, with no significant difference in other parameters.</p><p><b>CONCLUSION</b>The LC-MS determination method established in this experiment was so exclusive, accurate and sensitive that it is suitable for pharmacokinetic studies on extracts of HLXL and SI from L. chuanxion. The experiment results show that other ingredients of HLXL have noticeable effect on the absorption of SI in rat plasma.</p>
Subject(s)
Animals , Male , Rats , Administration, Oral , Area Under Curve , Benzofurans , Blood , Pharmacokinetics , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacokinetics , Metabolic Clearance Rate , Random Allocation , Rats, Sprague-DawleyABSTRACT
The purpose of the article is to apply a binary logistic model to analyze the major factors, which influence Chinese medicinal herb growers' willingness to use green pesticides by using survey data collected in Wenshan, Yunnan Province. The results indicate that, output per capita, average pesticide cost per mu, cognition of pesticide residues, expectations on Panax notoginseng prices, cognition of pesticides' effect of pests control, cognition of P. notoginseng prices of low pesticide residues have a significant influence on growers' willingness to use green pesticides. According to the analysis above, some proposals for enhancing Chinese medicinal herb growers' willingness to use green pesticides are put forward, such as, moving toward the intensive planting systems, fetching down the pieces of green pesticides, emphasizing and propagating the advantages of green pesticides, keeping the prices of Chinese medicinal herb running at steady rates.
Subject(s)
Adult , Female , Humans , Male , Young Adult , Attitude , China , Organic Agriculture , Economics , Workforce , Panax notoginseng , Pesticides , Economics , Plant Diseases , Plants, MedicinalABSTRACT
An HPLC method for the determination of 18alpha-glycyrrhetinic acid and 18beta-glycyrrhetinic acid in rat plasma was established, which was used subsequently to determine the pharmacokinetic profiles of both epimers of glycyrrhetinic acid in rats. alpha-glycyrrhetinic acid, beta-glycyrrhetinic acid, and a mixture of alpha-glycyrrhetinic and beta-glycyrrhetinic acids were administered to rats via gastric infusion. Blood samples were collected at different time intervals and extracted by liquid-liquid extraction. Separation was achieved by using a Kromasil C18 column (150 mm x 4.6 mm, 5 microm) with the mobile phase composed of acetonitrile--4 mmol x L(-1) ammonium acetate solution (46 : 54, v/v) at a flow rate of 1.0 mL x min(-1), and the detection wavelength was set at 250 nm. The pharmacokinetic parameters were calculated using the software DAS 2.0. In a combined administration, the main pharmacokinetic parameters of beta-glycyrrhetinic acid are significantly different from that of alpha-glycyrrhetinic acid (P < 0.05), while no significant difference was obtained when administrated individually. Compared to the single administration, significant differences (P < 0.05) on the values of AUC(0-t) and AUC(0-infinity) of beta-glycyrrhetinic acid were observed when this chemical was administrated together with alpha-glycyrrhetinic acid. In contrast, the pharmacokinetic parameters of alpha-glycyrrhetinic acid were not affected even under the co-administration. Here, a sensitive, specific, rapid and reproducible HPLC method was developed for the pharmacokinetic studies of alpha-glycyrrhetinic acid and beta-glycyrrhetinic acid in rat plasma.
Subject(s)
Animals , Male , Rats , Area Under Curve , Chromatography, High Pressure Liquid , Methods , Glycyrrhetinic Acid , Blood , Pharmacokinetics , Rats, Sprague-Dawley , StereoisomerismABSTRACT
With the excellent merits of wide analytical range, high sensitivity, small sample size, fast analysis speed, good repeatability, simple operation, low mobile phase consumption, as well as its capability of simultaneous isolation and identification, etc, mass spectrometry techniques have become widely used in the area of environmental science, energy chemical industry, biological medicine, and so on. This article reviews the application of mass spectrometry technology in biological sample analysis in the latest three years with the focus on the new applications in pharmacokinetics and bioequivalence, toxicokinetics, pharmacokinetic-pharmacodynamic, population pharmacokinetics, identification and fragmentation pathways of drugs and their metabolites and metabonomics to provide references for further study of biological sample analysis.
Subject(s)
Animals , Humans , Drug-Related Side Effects and Adverse Reactions , Mass Spectrometry , Methods , Metabolomics , Pharmaceutical Preparations , Chemistry , Pharmacokinetics , Pharmacology , Therapeutic EquivalencyABSTRACT
UPLC-MS-MS system was used for the identification of arbidol metabolites in the rat feces, urine and plasma samples. The system was so powerful a way with high ability of separation and analysis, based on both chromatography and mass properties. The isotope of Br was also a good indicator for metabolites finding. There were altogether 9 metabolites detected and identified, including 2 phase I biotransformation products and 7 phase II ones. It is concluded that arbidol mainly undergo metabolic reactions such as N-demethylation, S-oxidation, glucuronidation and sulfation in rats.
Subject(s)
Animals , Female , Male , Rats , Biotransformation , Chromatography, High Pressure Liquid , Feces , Chemistry , Indoles , Blood , Metabolism , Pharmacokinetics , Urine , Rats, Wistar , Tandem Mass SpectrometryABSTRACT
A LC-MS method was established for the determination of the protein binding rates of oleanolic acid in human plasma and serum albumin. The equilibrium dialysis combined with LC-MS to determine the total concentration in plasma and free drug concentration of oleanolic acid was carried out. The human plasma protein binding rates of oleanolic acid at three concentrations were 79.6%, 81.9% and 63.3%, respectively. The human serum albumin protein binding rates of oleanolic acid at three concentrations were 53.5%, 56.6% and 47.7%, respectively. The method is shown to be simple, accurate, sensitive and specific for the determination of biological samples. The protein binding rates in human plasma and serum albumin were of high strength.
Subject(s)
Humans , Chromatography, Liquid , Methods , Dialysis , Mass Spectrometry , Methods , Oleanolic Acid , Blood , Protein Binding , Sensitivity and Specificity , Serum Albumin , MetabolismABSTRACT
This paper is aimed to report the development of a method for the determination of the binding rate of plasma protein with salvianolic acid B. In vitro, equilibrium dialysis method was used to imitate the binding process between salvianolic acid B and plasma protein, in vivo, ultrafiltration method was used and the binding rate with HPLC was determined. Plasma samples were treated with methanol to precipitate the protein, and the buffer solution was directly determined after filtering. The calibration curve of the buffer solution was linear in the range of 0.5-20 microg mL(-1). The calibration curve of the plasma was linear in the range of 2-200 microg mL(-1). The extract recovery was 68.6%-81.9%. RSDs of intra- and inter-day precisions were all less than 8.5%. The binding rates of plasma protein with salvianolic acid B in vitro was 75.2% and in vivo was 92.1%. This paper shows the high binding power of salvianolic acid B to plasma protein with high sensitivity, good reproduction, simple management and fulfilling the requirement.
Subject(s)
Animals , Male , Rats , Benzofurans , Blood , Metabolism , Blood Proteins , Metabolism , Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Protein Binding , Rats, Sprague-Dawley , Reproducibility of Results , Salvia miltiorrhiza , Chemistry , Sensitivity and Specificity , Ultrafiltration , MethodsABSTRACT
To establish a method for simultaneous determination of dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid and pachymic acid in Poria, a RP-HPLC method detected by UV wavelengths switch had been developed, including 210 nm (48-55 min) for pachymic acid and 241 nm (0-48 min) for dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid, separately. The system consisting of a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) and a mixture of acetonitrile and 0.05% phosphate acid as the mobile phase was adopted; The flow rate was 1.0 mL x min(-1). The linear response range was 30.5-610.0 microg x mL(-1) (r = 0.999 6) for dehydrotumulosic acid, 12.66-253.2 microg x mL(-1) (r = 0.999 5) for polyporenic acid C, 2.99-59.7 microg x mL(-1) (r = 0.999 7) for 3-epi-dehydrotumulosic acid, 6.13-122.5 microg x mL(-1) (r = 0.999 5) for dehydropachymic acid and 11.3-226.0 microg x mL(-1) (r = 0.9995) for pachymic acid. The average recoveries of these compounds were 98.5% (RSD = 1.9%), 99.4% (RSD = 1.7%), 97.9% (RSD = 1.2%), 96.7% (RSD = 2.5%) and 97.9% (RSD = 2.3%), respectively. The method is simple, accurate and reproducible for quality control of Poria.