RESUMO
In this study, a temperature-sensitive molecularly imprinted polymer was prepared by using the bifunctional monomer with the critical phase transition characteristics. Infrared spectrometry, scanning electron microscopy, and specific surface area testing were used to characterize the polymers. Then, the recognizing properties of the polymers were studied. Based on the prepared smart polymers, an SPE-HPLC analytical method for the determination of quinolizidine alkaloids in the extracts of Sophora flavescens was established and verified. Finally, the smart polymers were applied to the enrichment of quinolizidine alkaloids in plant extracts. By changing the temperature and solvents of the solid phase extraction conditions, the extraction process can increase the concentration of quinolizidine alkaloids by 4.3 to 5.2 folds. The extraction process has mild conditions and less time consumption, avoiding the use of a large number of toxic reagents, which indicate that the extraction process are more efficient and environmentally friendly.
Assuntos
Alcaloides/análise , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/análise , Polímeros Molecularmente Impressos/química , Quinolizinas/análise , Extração em Fase Sólida/métodos , Alcaloides/isolamento & purificação , Medicamentos de Ervas Chinesas/isolamento & purificação , Polímeros Molecularmente Impressos/síntese química , Quinolizinas/isolamento & purificação , Extração em Fase Sólida/instrumentação , Sophora/química , MatrinasRESUMO
BACKGROUND: Sophorae tonkinensis Radix et Rhizoma is traditionally used for clearing away heat and toxic materials in China. PURPOSE: This study aims to observe the amelioration of Sophorae tonkinensis water extract (STR) against non-alcoholic fatty liver disease (NAFLD) and the engaged mechanism. METHODS: NAFLD was induced in mice fed by methionine and choline deficient (MCD) diet. Liver histological observation, Oil Red O, Masson's trichrome and F4/80 immunohistochemical staining were performed. Serum and liver biochemical parameters, hepatic gene and protein expression were detected. Cellular lipids accumulation in human normal liver l-02 and hepatoma HepRG cells were induced by 0.5 mM nonestesterified fatty acid (NEFA). The contents of matrine (MT) and oxymatrine (OMT) in STR were detected by using high-performance liquid chromatography (HPLC). Carnitine palmitoyltransferase 1A (CPT1A) expression and enzymatic activity were detected both in vivo and in vitro. RESULTS: Serum alanine/aspartate aminotransferase (ALT/AST) activity, hepatic malondialdehyde (MDA) content and liver histological observation showed that STR alleviated hepatocellular damage in mice fed with MCD diet. Hepatic triglyceride (TG), total cholesterol (TC) and NEFA amounts, and Oil Red O staining showed that STR reduced hepatic lipids accumulation in mice fed with MCD diet. STR and its main compounds including MT and OMT decreased NEFA-induced cellular lipids accumulation in hepatocytes. STR enhanced CPT1A activity both in vivo and in vitro. MT and OMT also enhanced cellular CPT1A activity in l-02 hepatocytes treated with NEFA. Moreover, the CTP1A inhibitor etomoxir (ETO) reduced the lipid-lowering activity provided by STR, MT or OMT in vitro. Liver myeloperoxidase (MPO) activity and hydroxyproline content, Masson's trichrome and F4/80 immunohistochemical staining, and hepatic mRNA expression of some molecules involved in regulating inflammation or fibrosis demonstrated that STR alleviated hepatic inflammation and liver fibrosis in mice fed with MCD diet. CONCLUSION: STR alleviated NAFLD by inhibiting hepatic inflammation and liver fibrosis, and reducing hepatic lipids accumulation through promoting fatty acids ß-oxidation by enhancing liver CPT1A activity. MT and OMT may be the main active compounds contributing to the lipid-lowering activity provided by STR.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Extratos Vegetais/farmacologia , Sophora/química , Alanina Transaminase/sangue , Alcaloides/análise , Animais , Linhagem Celular , Dieta/efeitos adversos , Medicamentos de Ervas Chinesas/química , Ácidos Graxos não Esterificados/metabolismo , Humanos , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/genética , Cirrose Hepática/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/metabolismo , Quinolizinas/análise , Triglicerídeos/sangue , Triglicerídeos/metabolismo , Água/química , MatrinasRESUMO
A novel micro matrix solid phase dispersion method was successfully used for the extraction of quaternary alkaloids in Phellodendri chinensis cortex. The elution of target compounds was accomplished with sodium hexanesulfonate as the eluent solvent. A neutral ion pair was formed between ion-pairing reagent and positively charged alkaloids in this process, which was beneficial for selectively extraction of polar alkaloids. Several parameters were optimized and the optimal conditions were listed as follows: silica gel as the sorbent, silica to sample mass ratio of 1:1, the grinding time of 1 min. The exhaustive elution of targets was achieved by 200 µL methanol/water (9:1) containing 150 mM sodium hexane sulfonate at pH 4.5. The method validation covered linearity, recovery, precision of intraday and interday, limits of detection, limits of quantitation, and repeatability. This established method was rapid, simple, environmentally friendly, and highly sensitive.
Assuntos
Berberina , Medicamentos de Ervas Chinesas/química , Quinolizinas , Extração em Fase Sólida/métodos , Berberina/análise , Berberina/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Limite de Detecção , Modelos Lineares , Phellodendron/química , Plantas Medicinais/química , Quinolizinas/análise , Quinolizinas/isolamento & purificação , Reprodutibilidade dos TestesRESUMO
We describe the screening of a set of cryptopleurine derivatives, namely thienoquinolizidine derivatives and (epi-)benzo analogs with bioactive phenanthroquinolizidine alkaloids that induce cytotoxic effects in the mouse lymphocytic leukemia cell line L1210. We used three variants of L1210 cells: i) parental cells (S) negative for P-glycoprotein (P-gp) expression; ii) P-glycoprotein positive cells (R), obtained by selection with vincristine; iii) P-glycoprotein positive cells (T), obtained by stable transfection with a human gene encoding P-glycoprotein. We identified the most effective derivative 11 with a median lethal concentration of ≈13 µM in all three L1210 cell variants. The analysis of the apoptosis/necrosis induced by derivative 11 revealed that cell death was the result of apoptosis with late apoptosis characteristics. Derivative 11 did not induce a strong alteration in the proportion of cells in the G1, S or G2/M phase of the cell cycle, but a strong increase in the number of S, R and T cells in the subG1 phase was detected. These findings indicated that we identified the most effective inducer of cell death, derivative 11, and this derivative effectively induced cell death in S, R and T cells at similar inhibitory concentrations independent of P-gp expression.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Apoptose/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Leucemia/metabolismo , Leucemia/patologia , Fenantrolinas/análise , Fenantrolinas/farmacologia , Quinolizinas/análise , Quinolizinas/farmacologia , Caspase 3/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ativação Enzimática , Humanos , Concentração Inibidora 50 , Modelos Moleculares , Fenantrolinas/química , Quinolizinas/química , Coloração e Rotulagem , Proteína X Associada a bcl-2/metabolismoRESUMO
To prepare standard decoction of Phellodendri Chinensis Cortex pieces, establish quality standards, and provide reference for evaluating the quality of formula granule of Phellodendri Chinensis Cortex. 15 batches Phellodendri Chinensis Cortex pieces in different levels of quality were collected from market. The standard extraction process was used to prepare the standard decoction, and then the transfer rate of the index components berberine and phellodendrine was calculated to measure dry extraction ratio and pH value. So the method for Phellodendri Chinensis Cortex's fingerprint and content determination of index components was established. The result revealed that the extraction ratio was from 12.81% to 19.41%, with an average value of 16.54%. The transfer rate of berberine was at the range of 36.4%-56.6%, with an average value of 48.9%. The transfer rate of Phellodendri Chinensis Cortex piece was at the range of 47.5%-83.3%, with an average value of 63.1%. The pH value was between 5.2 and 5.9. The Similarity Evaluation System for Chromatographic Fingerprint of traditional Chinese medicine TCM (2012A) was used to analyze and compare the fingerprint and the similarity between the fingerprint of formula granule of Phellodendri Chinensis Cortex and the reference fingerprint was over 0.9. The preparation method in this paper showed good precision, stability and repeatability in fingerprint analysis, suitable for quality evaluation of Phellodendri Chinensis Cortex pieces standard decoction.
Assuntos
Medicamentos de Ervas Chinesas/normas , Phellodendron/química , Controle de Qualidade , Berberina/análise , Quinolizinas/análiseRESUMO
A sensitive and rapid ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was developed for the simultaneous analysis of anemoside B4, phellodendrine, berberine, palmatine, obakunone, esculin, esculetin, toosendanin (IS1 of anemoside B4), tetrahydropalmatine (IS2 of phellodendrine, berberine, palmatine and obakunone) and scopoletin (IS3 of esculin and esculetin) and to compare the pharmacokinetics of these active ingredients in normal and ulcerative colitis rats. After methanol deproteinization, solvents were evaporated at 40°C under a gentle stream of nitrogen. Chromatography was performed using a C18 column with a gradient elution of 0.1% aqueous formic acid and acetonitrile at 0.4ml/min. Detection and measurement were performed on a 4000 QTRAP UPLC-MS/MS system from AB Sciex in the multiple reaction monitoring (MRM) mode. Phellodendrine, berberine, palmatine, obakunone, esculin, esculetin, tetrahydropalmatine (IS2) and scopoletin (IS3) were monitored under positive ionization conditions. Anemoside B4, and toosendanin (IS1) were monitored under negative ionization conditions. The optimized mass transition ion-pairs (m/z) were 1221.1/750.7 for anemoside B4, 343.2/193.2 for phellodendrine, 337.1/321.0 for berberine, 353.0/336.9 for palmatine, 455.1/161.1 for obakunone, 341.2/179.2 for esculin, 179.1/123.0 for esculetin, 573.4/531.4 for toosendanin (IS1), 356.2/192.2 for tetrahydropalmatine (IS2) and 193.0/133.1 for scopoletin (IS3).
Assuntos
Colite Ulcerativa/sangue , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/metabolismo , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Berberina/análise , Berberina/sangue , Alcaloides de Berberina/análise , Alcaloides de Berberina/sangue , Cromatografia Líquida de Alta Pressão/métodos , Esculina/análise , Esculina/sangue , Masculino , Quinolizinas/análise , Quinolizinas/sangue , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em Tandem/métodos , Umbeliferonas/análise , Umbeliferonas/sangueRESUMO
According to the research strategy of resource chemistry of Chinese medicinal materials and Chinese medicinal resources recycling utilization, this study intends to explore the potential resource-oriented utilization value of the seed of Sophora flavescens by contrasting with its kindred plant S. alopecuroides. This study established a rapid UPLC-Q-TOF-MS/MS and UPLC-TQ-MS/MS method to determine the alkaloids in the seed of S. flavescens. Results of UPLC-Q-TOF-MS/MS analysis showed that the alkaloids in the seed of S. flavescens were highly similar with S. alopecuroides.In the determination of 7 kinds of alkaloids, the total content was 11.203 and 15.506 mgâ¢g⻹ in the seed of S. flavescens and S. alopecuroides, respectively. The content of oxymatrine, oxysophocarpine and sophoridine is high in the seed of S. flavescens. The results indicated that the seeds of S. flavescens. could be an important material resource to obtain alkaloids.
Assuntos
Alcaloides/análise , Quinolizinas/análise , Sementes/química , Sophora/química , China , Compostos Fitoquímicos/análise , Espectrometria de Massas em TandemRESUMO
This study intends to explore the potential resource-orientedutilization value of the flower of Sophora flavescents by analyzing alkaloids and flavonoids in the flower of S. flavescens from Shanxi province. This study established a rapid UPLC-TQ-MS/MS method that is used for determination of seven alkaloids and seven flavonoids in the flower of S.flavescens. The different florescences all have the seven detected alkaloids such as cytisine, oxy-matrine, oxy-sophocarpine, sophoridine, N-methylcytisine, matrine, sophocarpine.The total contents of detected alkaloids are as follows: flower buds 1.47%, primal flowers 1.34%, full bloomed flowers 1.17%, faded flowers 1.01%. The top three contents of alkaloids are N-methylcytisine , oxy-sophocarpine and oxymatrine, accounting for about 83% of the total amount of detected alkaloids. All the samples in different florescences have the seven detected flavonoids such as rutin, luteolin, quercetin, isoquercitrin, trifolirhizin, kurarinone, and kushenol I. The total contents of detected alkaloids are as follows: flower buds 495.2 µgâ¢g⻹, primal flowers 313.7 µgâ¢g⻹, faded flowers 224.2 µgâ¢g⻹, full bloomed flowers 193.0 µgâ¢g⻹. The content of luteolinis relatively higher than other detected flavonoids, accounting for about 89%-94% of the total amount of detected flavonoids. The results indicated that the flower of S.flavescens could be an important material resource to obtain the resourceful alkaloids. This result can provide scientific basis for resource-oriented utilization and industrial development of the flower of S. flavescens.
Assuntos
Alcaloides/análise , Flavonoides/análise , Flores/química , Quinolizinas/análise , Sophora/química , China , Cromatografia Líquida de Alta Pressão , Compostos Fitoquímicos/análise , Espectrometria de Massas em TandemRESUMO
A reversed phase high-performance liquid chromatographic method (HPLC/MS-MS) has been developed and validated for detection of alkaloids matrine and oxymatrine in fertilizer with labeled enhancer plant defense activities. The analytical method was validated statistically. The results show a strong matrix effect, requiring quantification by standard addition method. The regression lines showed r(2) > 0.994. Recoveries ranging from 97 to 104% were obtained for the fortification level of 0.01% wt wt(-1) and the relative standard deviations ranged from 3 to 4% (n = 10). The limits of detection were below 0.0001% wt wt(-1), while the limits of quantification did not exceed 0.0004% wt wt(-1). The method is currently applied in ICQRF Laboratory of Catania on fertilized and corroborant plant extract collected in the Italian market in the frame of MIPAAF institutional quality control activity, with the aim to dectect these unpermitted active substances.
Assuntos
Alcaloides/análise , Cromatografia Líquida de Alta Pressão/métodos , Fertilizantes/análise , Extratos Vegetais/análise , Quinolizinas/análise , Sophora/química , Espectrometria de Massas em Tandem/métodos , MatrinasRESUMO
Alkaloid contents of leaf and seed samples of eight species of Sophora native to New Zealand, plus Sophora cassioides from Chile are reported. Fifty-six leaf and forty-two seed samples were analysed for alkaloid content by proton nuclear magnetic resonance spectroscopy, which showed major alkaloids as cytisine, N-methyl cytisine and matrine. GC analyses quantified these and identified further alkaloid components. The alkaloids identified were cytisine, sparteine, and matrine-types common to Sophora from other regions of the world. Cytisine, N-methyl cytisine, and matrine were generally the most abundant alkaloids across all species with seeds containing the highest concentrations of alkaloids. However, there was no clear taxonomic grouping based on alkaloid composition. A quantitative analysis of various parts of two Sophora microphylla trees showed that the seeds were the richest source of alkaloids (total 0.4-0.5% DM), followed by leaf and twig (0.1-0.3%) and then bark (0.04-0.06%), with only low amounts (<0.02%) found in the roots. This study represents the most comprehensive phytochemical investigation of New Zealand Sophora species to date and presents data for three species of Sophora for which no prior chemistry has been reported.
Assuntos
Alcaloides/análise , Sophora/química , Alcaloides/química , Alcaloides/metabolismo , Azocinas/análise , Chile , Medicamentos de Ervas Chinesas/química , Estrutura Molecular , Nova Zelândia , Folhas de Planta/química , Raízes de Plantas/química , Quinolizinas/análise , Sementes/química , Sophora/genética , MatrinasRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Zeng-Sheng-Ping (ZSP), also called antitumor B, is a marketed Chinese traditional medicine used for cancer prevention. AIM OF THE STUDY: Currently, for the quality control of Chinese traditional medicines, marker compounds are not selected based on bioactivities and pharmaceutical behaviors in most of the cases. Therefore, even if the "quality" of the medicine is controlled, the pharmacological effect could still be inconsistent. The aim of this study is to establish an activity and absorption-based platform to select marker compound(s) for the quality control of Chinese traditional medicines. MATERIALS AND METHODS: We used ZSP as a reference Chinese traditional medicine to establish the platform. Activity guided fractionation approach was used to purify the major components from ZSP. NMR and MS spectra were used to elucidate the structure of the isolated compounds. MTT assay against oral carcinoma cell line (SCC2095) was performed to evaluate the activities. UPLC-MS/MS was used to quantify the pure compounds in ZSP and the active fraction. The permeabilities of the identified compounds were evaluated in the Caco-2 cell culture model. The intracellular accumulation of the isolated compounds was evaluated in the SCC2095 cells. RESULTS: The major compounds were identified from ZSP. The contents, anti-proliferation activities, permeabilities, and intracellular accumulations of these compounds were also evaluated. The structure of these purified compounds were identified by comparing the NMR and MS data with those of references as rutaevine (1), limonin (2), evodol (3), obacunone (4), fraxinellone (5), dictamnine (6), maackiain (7), trifolirhizin (8), and matrine (9). The IC50 of compounds 5, 6, and 7 against SCC2095 cells were significantly lower than that of ZSP. The uptake permeability of compounds 5, 6, and 7 were 2.58 ± 0.3 × 10(-5), 4.33 ± 0.5 × 10(-5), and 4.27 ± 0.8 × 10(-5) respectively in the Caco-2 cell culture model. The intracellular concentrations of these compounds showed that compounds 5, 6, and 7 were significantly accumulated inside the cells. CONCLUSION: Based on the activity against oral carcinoma cell line as well as the absorption permeability, compound 5, 6, and 7 are selected as quality control markers for ZSP. An activity and absorption-based platform was established and successfully used for the quality control of ZSP.
Assuntos
Proliferação de Células/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Medicina Tradicional Chinesa/normas , Controle de Qualidade , Alcaloides/análise , Alcaloides/isolamento & purificação , Benzofuranos/análise , Benzofuranos/isolamento & purificação , Benzoxepinas/análise , Benzoxepinas/isolamento & purificação , Linhagem Celular Tumoral , Glucosídeos/análise , Glucosídeos/isolamento & purificação , Glucosídeos/farmacocinética , Compostos Heterocíclicos de 4 ou mais Anéis/análise , Compostos Heterocíclicos de 4 ou mais Anéis/isolamento & purificação , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Humanos , Limoninas/análise , Limoninas/isolamento & purificação , Permeabilidade , Pterocarpanos/análise , Pterocarpanos/isolamento & purificação , Pterocarpanos/farmacocinética , Quinolinas/análise , Quinolinas/isolamento & purificação , Quinolinas/farmacocinética , Quinolizinas/análise , Quinolizinas/isolamento & purificação , Triterpenos/análise , Triterpenos/isolamento & purificação , MatrinasRESUMO
OBJECTIVE: To optimize the preparation technology of Shangke Jiefu lotion. METHODS: The extraction process was optimized by orthogonal test, with water addition, extraction times and time used for extraction as factors of investigation. In refined process test, alcohol precipitation concentration, time, and the relative density of extract were studied. Each factor had three levels. The content of sophorcarpidine and the yield of dry extract were used as the evaluation indexes. The content of sophorcarpidine was determined by HPLC, and dry extract rates were determined by drying method. RESULTS: The best extraction condition was as follows: the amount of water was 10 times of the medicinal materials, the decoction duration was 2 h and for 3 times. The optimum purification process was: alcohol precipitation concentration was 50%, time was 15 hours, relative density of extract was 1.05 g/mL. CONCLUSION: The optimized preparation technology of Shangke Jiefu lotion is stable, feasible and convenient. It provides a theoretical basis for standardized production.
Assuntos
Alcaloides/análise , Fracionamento Químico/métodos , Desinfetantes/isolamento & purificação , Medicamentos de Ervas Chinesas/isolamento & purificação , Plantas Medicinais/química , Quinolizinas/análise , Cromatografia Líquida de Alta Pressão , Desinfetantes/química , Medicamentos de Ervas Chinesas/química , Temperatura Alta , Controle de Qualidade , Sophora/química , Fatores de Tempo , Água/química , MatrinasRESUMO
OBJECTIVE: To establish a new method for quality evaluation and validate its feasibilities by simultaneous quantitative assay of five alkaloids in Sophora flavescens. METHOD: The new quality evaluation method, quantitative analysis of multi-components by single marker (QAMS), was established and validated with S. flavescens. Five main alkaloids, oxymatrine, sophocarpine, matrine, oxysophocarpine and sophoridine, were selected as analytes to evaluate the quality of rhizome of S. flavescens, and the relative correction factor has good repeatibility. Their contents in 21 batches of samples, collected from different areas, were determined by both external standard method and QAMS. The method was evaluated by comparison of the quantitative results between external standard method and QAMS. RESULT: No significant differences were found in the quantitative results of five alkaloids in 21 batches of S. flavescens determined by external standard method and QAMS. CONCLUSION: It is feasible and suitable to evaluate the quality of rhizome of S. flavescens by QAMS.
Assuntos
Alcaloides/análise , Sophora/química , Quinolizinas/análise , MatrinasRESUMO
In order to improve the resolution and peak shape, 1-butyl-3-methylimidazolium tetrafluoroborate ( [ BMIM] BF4 ) was used as mobile phase additive to develop an HPLC method for the determination of four main alkaloids in Compound Kushen Injection. An Agilent TC-C18 column was used with the mobile phase of methanol-0.1% phosphoric acid solution (containing 2. 2 x 10(4) mol/L [BMIM] BF4 ) (5: 95, v/v) , flow rate of 1.0 mL/min, column temperature of 30 degree C, and injection volume of 20 micro L, detected at 205 nm. The calibration curves of matrine, sophoridine, oxymatrine and oxysophocarpine were in good linearity over the ranges of 25.8 -155.0 mg/L, 40.0 -240.0 mg/L, 21.7 - 130.0 mg/L and 37.5 -225.0 mg/L, respectively, with correlation coefficients above 0. 999 0. The mean recoveries (n = 9) were between 96. 2% and 98.9%. Ionic liquid, [BMIM] BF4, can obviously improve the resolution and peak shape. The method is simple, rapid, and reliable, and can be used for the determination of alkaloids in Sophora flavescens Ait in Compound Kushen Injection.
Assuntos
Alcaloides/análise , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Líquidos Iônicos/química , Combinação de Medicamentos , Quinolizinas/análise , MatrinasRESUMO
A Cleanert Alumina-N-SPE column (0.5 g/6 mL) chromatograpy with 5 mL of chloroform-methanol (7: 3) as eluent, instead of aluminum oxide column (100-200 mesh, 5 g, 1 cm) chromatograpy eluted successively with chloroform and the chloroform-methanol (7:3) (20 mL each), was applied to enrich matrine and oxymatrine in Sophora flavescens. Also, the optimization of the HPLC determination conditions with acetonitrile-ethanol absolute-3% phosphoric acid solution (84: 6: 10) as mobile phase, instead of acetonitrile-ethanol absolute -3% Phosphoric acid solution (80: 10: 10) recorded in Chinese Pharmacopoeia 2010 Edition, was more suitable for determination of matrine and oxymatrine in S. flavescens. This method has advantage of reducing sample handling time and solvent volume and increasing the accuracy and feasibility, which can simplify the procedure for determination of matrine and oxymatrine in S. flavescens.
Assuntos
Alcaloides/isolamento & purificação , Medicamentos de Ervas Chinesas/isolamento & purificação , Quinolizinas/isolamento & purificação , Extração em Fase Sólida/métodos , Sophora/química , Alcaloides/análise , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/análise , Quinolizinas/análise , MatrinasRESUMO
OBJECTIVE: To study the effect of transplantation on the growth and oxymatrine content of Sophora flavescens and provide foundation for popularization and cultivation of Sophora flavescens in South China. METHODS: Sophora flavescens which was usually planted in North China and Northwest China was planted in a non-traditional location, Zhongshan city, Guangdong Province in South China to test its adaptability. The growth characters, such as plant height, leaf area, dry weight of root, diameter and length of root and so on were measured from 2010 to 2012. The oxymatrine content of one-year old and two-year old root of Sophora flavescens were determined by HPLC. Nine major growth indexes for one-year old Sophora flavescens were comprehensively analyzed and evaluated by the methods of weighted gray relational and hierarchy evaluation of fuzzy mathematics. RESULTS: The weighted relevancy of introduced and reference cultivars was 0.8545. The introduced cultivar was rather adaptable to the geography environment in Zhongshan. Its quality was very close to the reference cultivars. Oxymatrine content of root of one-year old and two-year old Sophora flavescens was 13.2784 mg/g (as much as origin) and 16.4779 mg/g (less than origin 28.67%), respectively. These were 10.65% and 37.32% higher than the quality standard which were set up in the Chinese Pharmacopoeia (2010 edition). CONCLUSION: Sophora flavescens performs quite well in the newly introducing region. It is suitable to be cultivated and extended in South China.
Assuntos
Alcaloides/análise , Cromatografia Líquida de Alta Pressão/métodos , Quinolizinas/análise , Sophora/química , Sophora/crescimento & desenvolvimento , China , Clima , Ecossistema , Fertilizantes , Folhas de Planta/anatomia & histologia , Folhas de Planta/química , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/química , Raízes de Plantas/crescimento & desenvolvimento , Plantas Medicinais/química , Plantas Medicinais/crescimento & desenvolvimento , Estações do Ano , Sophora/anatomia & histologiaRESUMO
Cell membrane chromatography (CMC) is a biological affinity chromatographic method using specific cell membrane as stationary phase. It has been proved to be a practical tool for investigating binding interactions between drugs and membrane receptors. In this study, a novel comprehensive two-dimensional (2D) chromatography approach was established for screening anti-tumor components from herbal medicines (HMs). HepG2/CMC model was first developed and applied as the first dimensional column. Using an automatic ten-port switching valve equipped with two sample loops, the fractions of the first-dimension were introduced in the second-dimension consists of a monolithic column and a time-of-flight mass spectrometry (TOFMS) with high resolving ability. Based on the stability, selectivity and suitability assays of the HepG2/CMC/monolithic column/TOFMS system, berberine (BBR) and tetrahydropalmatine (THP) from Cortex phellodendri amurensis, oxymatrine and matrine from Radix sophorae ï¬avescentis were screened and identified as potential active components. The competitive displacement assay suggested that the four components could act on epidermal growth factor receptor region on the HepG2 cell membrane in similar manner of gefitinib. Furthermore, their inhibiting effects on cell proliferation in vitro were also confirmed and, BBR and THP showed concentration dependently inhibitory ability on HepG2 cell proliferation (p<0.05). The result demonstrated that the proposed comprehensive 2D HepG2/CMC/monolithic column/TOFMS system has the advantages of strong recognition and rapid analysis abilities for the total screening procedure, which will be selectable and practical in drug discovery from complex HM samples and can also be applied to other biochromatography models.
Assuntos
Antineoplásicos/química , Cromatografia Líquida/métodos , Medicamentos de Ervas Chinesas/química , Espectrometria de Massas/métodos , Alcaloides/análise , Alcaloides/química , Alcaloides/farmacologia , Antineoplásicos/análise , Antineoplásicos/farmacologia , Berberina/análise , Berberina/química , Berberina/farmacologia , Alcaloides de Berberina/análise , Alcaloides de Berberina/química , Alcaloides de Berberina/farmacologia , Proliferação de Células/efeitos dos fármacos , Receptores ErbB/antagonistas & inibidores , Gefitinibe , Células Hep G2 , Humanos , Quinazolinas/análise , Quinazolinas/química , Quinazolinas/farmacologia , Quinolizinas/análise , Quinolizinas/química , Quinolizinas/farmacologia , Reprodutibilidade dos Testes , MatrinasRESUMO
A sensitive and accurate method for the simultaneous determination of five alkaloids, namely 9α-hydroxymatrine (M1), matrine (M2), sophoridine (M3), oxymatrine (M4), alopecurin A (M5) in different parts (seed, legume, stem, and root) and different harvest times of Sophora alopecuroides L. was developed by high performance liquid chromatography (HPLC) with photodiode array detector (PDA) for the first time. The separation by gradient elution was achieved on Scienhome Kromasil C(18) (4.6×250 mm, 5 µm) column at 30°C with acetonitrile (A)/0.1% phosphatic acid+0.1% triethylamine (B) as the mobile phase. The detection wavelength was 205 nm. The optimized method provided a good linear relation (r≥0.9993 for all the target compounds), satisfactory precision (RSD values less than 2.3%) and good recovery (96.4-103.6%). The limits of detection ranged between 0.11×10(-3) and 4.70×10(-3) µg for the different analytes. The method was successfully applied to analysis and quality control of alkaloid extracts from the traditional Chinese herbal drugs of S. alopecuroides L.
Assuntos
Alcaloides/análise , Extratos Vegetais/química , Quinolizinas/análise , Sophora/química , Alcaloides/metabolismo , Cromatografia Líquida de Alta Pressão , Estabilidade de Medicamentos , Modelos Lineares , Especificidade de Órgãos , Componentes Aéreos da Planta/química , Componentes Aéreos da Planta/crescimento & desenvolvimento , Extratos Vegetais/metabolismo , Quinolizinas/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Sophora/crescimento & desenvolvimento , Fatores de TempoRESUMO
The aim of the research was to develop a method for quantitative determination of cytisine in Spartium junceum L. We used the above-ground parts of plants. In developing a method of analysis we used the method of 3-phase extraction. In this case the best results were obtained in the system: chopped raw material - water solution of ammonia - chloroform. In this case, the amount of alkaloids extracted almost entirely from the plant and goes into the chloroform phase. Evaluation of the results was carried out by the validation. The method for determination of cytisine in raw product was proposed. The method comprises the following steps-extraction of raw materials extracting chloroform phase and its evaporation, the translation of solids in methanol, the chromatographic separation cytisine and its fixation of the spectrophotometer method. The method is reproducible, has the required accuracy, is easy to analysis (less than 9 hours).
Assuntos
Alcaloides/análise , Alcaloides/química , Spartium/química , Alcaloides/isolamento & purificação , Azocinas/análise , Azocinas/química , Azocinas/isolamento & purificação , Clorofórmio/química , Cromatografia/métodos , República da Geórgia , Metanol/química , Extratos Vegetais/análise , Extratos Vegetais/química , Quinolizinas/análise , Quinolizinas/química , Quinolizinas/isolamento & purificaçãoRESUMO
OBJECTIVE: To establish a method for determination of 7 alkaloid in Herba Sophorae Alopecuroidis by HPLC. METHOD: X-Brige C18 (4.6 mm x 200 mm, 5 microm) column was used with acetonitriles-0.05 mol x L(-1) KH2PO4 solution (2.0 mL x L(-1) triethylamine) with gradient elution as the mobile phase and 1.0 mL x min(-1) as the flow rate. The detection wavelength was 205 nm. RESULT: Aloperin curve was linear in the range from 20.66 to 103.32 microg (r = 0.998 8) and the average recovery was 97.12% (RSD 7. 3%); sophoridine curve was linear in the range from 22.82 to 114.12 microg (r = 0.999 7) and the average recovery was 97.47% (RSD 3.0%); oxymatrine curve was linear in the range from 25.10 to 125.52 microg (r = 0.999 1) and the average recovery was 96.21% (RSD 4.5%); oxysophocarpine curve was linear in the range from 23.88 to 119.40 microg (r = 0.997 5) and the average recovery was 94. 64% (RSD 5.2%); matrine curve was linear in the range from 5.00 to 24.99 microg (r = 0.998 6) and the average recovery was 98.04% (RSD 5.4%); sophocarping curve was linear in the range from 4.69 to 23.46 microg (r = 0.999 6) and the average recovery was 96.24 (RSD 5.8%); lehmannine curve was linear in the range from 4.60 to 23.01 microg (r = 0.997 8) and the average recovery was 101.31% (RSD 4.3%). CONCLUSION: The method is accurate, simple and feasible. It can be used as a quality evaluation in Herba S. Alopecuroidis.