Exploration of nanofiltration conditions for separation of alcohol precipitation liquid during preparation of Reduning Injection by Box-Behnken design combined with flux attenuation / 中国中药杂志

This study employed Box-Behnken design combined with flux attenuation to explore the nanofiltration conditions for separation of alcohol precipitation liquid during the preparation of Reduning Injection and discussed the applicability of nanofiltration in the separation of the liquid with high-concentration ethanol. The effects of nanofiltration molecular weight cut-off(MWCO) and pH on the rejection of chlorogenic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid were consistent with the principles of pore size sieving and charge effect, respectively. The rejection of the three phenolic acids was reduced by concentration polarization effect caused by trans-membrane pressure(TMP). The swelling of membrane surface decreased the pore size and membrane flux for effective separation. Chlorogenic acid and 4,5-dicaffeoylquinic acid were more sensitive to pH and ethanol concentration than 3,5-dicaffeoylquinic acid. A certain correlation existed between the compound structure and the separation factors of nanofiltration, and the separation rules were associated with the comprehensive effect of charge effect, pore size sieving, concentration polarization, steric hindrance and so on.

Study on law of quality value transmitting of Artemisiae Scopariae Herba standard decoction / 中草药

Objective: To study the law of quality value transmitting of Artemisiae Scopariae Herba (ASH) standard decoction. Methods: Fifteen batches of ASH standard decoction were prepared. Fingerprints of these 15 batches standard decoction and its raw pieces were determined by HPLC, the control fingerprints were established, the common peaks were calibrated and the similarity was evaluated. Components of common peaks were identified by Q-TOF/MS and the contents of components confirmed by reference substance were determined. The common peaks transfer number, peak area ratio, index components transfer rate and extraction rate were used to analyze the quality value transfer rule of standard decoction. Results: The similarity of fingerprints of ASH and its standard decoction were greater than 0.9, 16 and 15 common peaks were calibrated respectively, and the transfer rate of the common peaks number was 93.75%. Thirteen components of the common peaks were confirmed by reference substance, including 1-caffeoylquinic acid, neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, p-hydroxyacetophenone, 1,3-dicaffeoylquinic acid, rutin, hyperoside, isoquercetin, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid. Caffeoylquinic acid glucoside and a di-C-glycoside named apigenin 6,8-di-C-β-D-glucoside were first detected in ASH. The ratio of the common peaks area of ASH standard decoction to that of ASH raw pieces showed a good positive linear relationship with the components transfer rate. The average transfer rates of neochlorogenic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid were more than 100%, while the average transfer rates of chlorogenic acid and 3,5-dicaffeoylquinic acid were only 47.59% and 22.33% respectively, suggesting that the organic acid components may be transformed into each other during the preparation of standard decoction, and part of chlorogenic acid may be transformed into cryptochlorogenic acid and neochlorogenic acid; 3,5-dicaffeoylquinic acid may be partially converted into 1,3-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid. The average transfer rates of rutin and hyperoside were 31.36% and 28.36%, respectively, and that of other components were between 50% and 70%. The average extraction rate of standard decoction was 19.76%. Conclusion: The laws of quality value transmitting of ASH standard decoction revealed in this study laid a foundation for the establishment of the material reference of ASH formula granules and the classic prescriptions containing ASH.

Simultaneous determination of five active components in Fangshu Qingre mixture by RP-HPLC / 药学实践杂志

Objective To establish a method for simultaneous determination of chlorogenic acid, luteolin-7-O-β-D-glucoside,3,5-dicaffeoylquinic acid, linarin and pogostone in Fangshu Qingre mixture by RP-HPLC. Methods ZORBAX-SB-C18 column(4.6 mm×250 mm, 5 μm) was used as the chromatographic column.The mobile phase was 0.2% formic acid water solution(A) and acetonitrile solution (B)with a gradient elution mode. The flow rate was 1.0 ml/min.The detective wavelength was 327 nm. The column temperature was 30 ℃. Results There were good linear relationships in the determination of chlorogenic acid, luteolin-7-O-β-D-glucoside, 3,5-dicaffeoylquinic acid, linarin and pogostone (r≥0.999 6), with the average recovery rate of 102.03%(1.63%), 102.38%(1.51%), 102.39%(1.23%), 103.14%(1.87%) and 104.01%(2.33%). Conclusion The method was simple and stable with a good reproducibility, which could be used as a quality control method for active compotents in Fangshu Qingre mixture.

Preliminary prediction and analysis of Q-marker of multi-original Tibetan medicine meiduoluomi / 中草药

Objective Based on the concept of quality marker (Q-marker), a chemical method was developed to evaluate the quality of the multi-original medicinal material “Meiduoluomi” and predict its Q-marker. Methods Firstly, the Q-marker were preliminary predicted based on the relationship between genetic relationship and biosynthesis pathway, pharmacological activity and chemical composition. Secondly, the main active components were determined by UHPLC. The analysis was performed using ACQUITY UHPLC HSS C18 column (100 mm × 2.1 mm, 1.8 μm); Mobile phase was acetonitrile (A)-0.3% acetic acid water (B) with gradient elution; The flow rate was 0.2 mL/min; The column temperature was 35 ℃; The detection wavelength was 320 nm. Results A new method for simultaneous determining five compounds of meiduoluomi was established, including chlorogenic acid, caffeic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid. The linear relationship of them was good at the range of 9.60-480.00, 10.27-513.50, 10.08-504.03, 9.64-482.80, and 3.82-380.24 mg/L, respectively; And the average recovery rates were 102.43%, 99.78%, 99.39%, 103.12%, and 101.83%, respectively. The five components were detected in each sample and the total content of the five components was more than 10% in the herbal medicine. It was suggested that the five components can be used as a Q-marker of meiduoluomi for quality control. Conclusion The coffeic acid components are scientific and reasonable to be considered as a Q-marker of meiduoluomi. The developed UHPLC method can be used for the quality control of meiduoluomi. This study provides new idea for the rational evaluation of multi-original medicinal material based on Q-marker.

Induction of mice death by toxicity components from large-dose Coreopsis tinctoria extract / 中草药

Objective: By comparing the acute toxicity of different extracts from Coreopsis tinctoria on mice, combined with the HPLC fingerprint and multiple linear regression to analyze the element which plays the most important role in causing the death of mice, and to provide the safety data for improving the extraction technology. Methods: To measure the maximum dose and maximal tolerance dose (MTD) of all the extracts, to measure the median lethal dose (LD50) by Bliss, and to record the death and weight changes; To measure the fingerprints of the extracts by HPLC, and to determine the element which mostly induced the death of mice by analyzing the absorption peak of the extracts by HPLC fingerprint with multiple linear regression. Results: The extracts include aqueous extract by spray drying (SD), aqueous extract by vacuum drying (VD) process, ethanol extract (ETE), ethyl acetate extracted component (AC), and the ethyl acetate extracted residuum (AR). Among those extracts, the maximum dose of SD and AR is 36 g/kg, the MTD of the VD is 26 g/kg, the LD50 (95% confidence limits) of ETE and AC are 19.565 (17.558-21.734) g/kg and 16.414 (13.987-34.725) g/kg, respectively; Under the high dose situation, 3,5-dicaffeoylquinic acid properly is the component which mostly contributes to the death of mice. Conclusion: Under the high dose situation, the ETE and AC will lead the death, and 3,5-dicaffeoylquinic acid properly is the component which mostly contributes to the death of mice.

Dynamic changes of phenolic acids and anthraquinones in aerial parts of Xanthium sibiricum from different collection periods / 中草药

Objective: To analyze the dynamic changes of phenolic acids and anthraquinones from the aerial parts of Xanthium sibiricum (Xanthii Herba) in different collection periods. Methods: To establish an HPLC method for simultaneous determination of chlorogenic acid, neochlorogenic acid, protocatechualdehyde, protocatechuic acid, cryptochlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, ferulic acid, 3,4- dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, aloeemodin, emodin, and chrysophanol in the aerial parts of X. sibiricum. Results: The contents of phenolic acids and anthraquinones in different harvest time showed dynamic changes. In mid July, the content of total phenolic acids was higher. The higher content of total anthraquinones was in late July. The contents of five total phenolic acids were higher in mid July, such as chlorogenic acid, protocatechuic aldehyde, ferulic acid, 3,5-dicaffeoylquinic acid, and 1,3-dicaffeoylquinic acid. The content of chlorogenic acid was higher in late June, and the rest of five phenolic acids were higher in mid July. Conclusion: This can provide the basis for determining the suitable harvest time of X. sibiricum.

Chemical constituents from whole plant of Gynura procumbens (I) / 中草药

Objective: To investigate the chemical constituents from the whole plants of Gynura procumbens. Methods: The chemical constituents were isolated and purified by repeated silica gel column chromatography, Sephadex LH-20 gel column chromatography, medium pressure column chromatography, and semi-preparative HPLC, and their structures were elucidated by chemical properties and spectroscopic analyses. Results: Sixteen compounds were identified to be quercetin (1), apigenin (2), luteolin (3), kaempferol (4), astragaline (5), kaempferol-5-O-(6″-O-acetyl)-β-D-glucopyranoside (6), negletein (7), 4-methoxycinnamic acid (8), benzyl-O-β-D-glucopyranoside (9), 2-phenylethyl-O-β-D-glucopyranoside (10), 3,5-dicaffeoylquinic acid methyl ester (11), 3,5-dicaffeoylquinic acid ethyl ester (12), 3,4-dicaffeoylquinic acid methyl ester (13), 4,5-dicaffeoylquinic acid methyl ester (14), protocatechuic acid (15), and eugenol glucoside (16). Conclusion: Compounds 7, 8, 12, 14, and 16 are obtained from the plants in Gynura Cass. for the first time, and compounds 3, 6, 9-11 and 13 are obtained from this plant for the first time.

Improvement of different processing technologies of Xanthii Fructus / 中草药

Objective: To compare the different processed products of Xanthii Fructus and improve the processing technology of Xanthii Fructus. Methods: Using 2015 version of Chinese Pharmacopoeia to determine the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid as well as toxic ingredients, such as carboxyatractyloside and atractyloside in raw products, stir-fried and sand fried active ingredients of Xanthii Fructus; The fingerprint of Xanthii Fructus samples was established, and the Chinese Pharmacopoeia Commission promulgated the chromatographic fingerprint similarity evaluation system software (version: 2004A) for the similarity evaluation; SPSS 19.0 software analyzed cluster analysis and SIMCAI 13.0 software analyzed partial least squares-discriminant analysis (PLS-DA). Results: The results showed that the active ingredient content of improved processing technology was high and toxic constituents content was low; The similarity of the 19 samples was greater than 0.98; Cluster analysis showed that the samples might be roughly classified into four groups; PLS-DA analysis showed that different samples can be distinguished. Conclusion: The established analysis method of Xanthii Fructus has a good reproducibility verifies the feasibility of improved processing technology, and provides a scientific basis for quality control of Xanthii Fructus.

Research on quality control method for Juhuang Oral Liquid / 中草药

Objective: To establish the quality control method for Juhuang Oral Liquid (JOL) based on the qualitative and quantitative analysis methods. Methods: TLC was used to identify Lycii Fructus and Polygonati Rhizoma in JOL. Phenol-sulfuric acid method was used for the detemination of total polysaccharide, HPLC was used to simultaneously determine the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, coffeic acid, galuteolin, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid. The Odyssil C18 (250 mm × 4.6 mm, 5 μm) column was used. The mobile phase was acetonitrile-0.05% phosphoric acid solution in gradient elution, at the flow rate of 1.0 mL/min, the detection wavelength was set at 325 nm. Results: The identified characteristics of Lycii Fructus and Polygonati Rhizoma by TLC were distinct and the spots were clear, without interference and easy to recognize. When phenol-sulfuric acid method was used to determinate the total polysaccharide content, the regression equation was A=0.057 8 C + 0.032 5 (r=0.999 1, n=8). The linear with its absorbance in the range of 26.15-196.13 mg/mL, the average recoveries was 99.82%. The average content of the total polysaccharide in JOL from six batches is 157.74-166.49 mg/mL. When RP-HPLC was used to determine the eight chemical components, the compounds showed a good linearity(r ≥ 0.999 8) in the determination range. The average recoveries were between 99.18%-101.06% with RSDs between 0.79%-1.91%. The average content from the six batches showed neochlorogenic acid 102.9-142.6 μg/mL, chlorogenic acid 488.6-563.8 μg/mL, cryptochlorogenic acid 58.9-71.6 μg/mL, coffeic acid 240.7-326.1 μg/mL, galuteolin 228.6-302.7 μg/mL, 3,4-dicaffeoylquinic acid 398.0-485.4 μg/mL, 3,5-dicaffeoylquinic acid 184.1-203.1 μg/mL, and 4,5-dicaffeoylquinic acid 476.2-561.8 μg/mL. Conclusion: TLC identification method is simple. The determination method of total polysaccharide and chemical components is simple, accurate, and with a good repeatability. It can be used for the quality control of JOL.

Quality comparison on Farfarae Flos from various origins by UPLC based on multi-components content determination method / 中草药

Objective: To establish a UPLC method for simultaneously determining 12 components and evaluating the quality of Farfarae Flos from various origins. Methods: The UPLC method was achieved by BEH C18 column (100 mm × 2.1 mm, 1.7 μm) using a mobile phase made up of acetonitrile (A)-0.03% trifluoroacetic acid in water (B). The flow rate and detection wavelength were 0.5 mL/min and 254 nm, respectively.The column temperature was 40 ℃. Coefficients of variation (CV) were calculated, then PCA and HCA were applied to analyzing Farfarae Flos from different origins. Results: The method for content determination was in agreement with methodological requirements. The results showed that CV values of different components differed greatly, and the hyperoside showed the largest CV value at 0.91. PCA and HCA analysis revealed the wild and cultivated Farfarae Flos were different, and the wild samples contained more chlorogenic acid, caffeic acid, rutin, hyperoside, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic, and tussilagone. Conclusion: There exist the significant differences among Farfarae Flos from various origins, and their CV values are also different. In addition, the wild samples also differ from those of the cultivated ones. To guarantee the safety and effectiveness in clinical, the impact of chemical differences of different Farfarae Flos on the clinical efficacy should be further studied.

Simultaneous determination of 17 major components of Shuang-Huang-Lian Oral Liquid by UPLC-MS/MS / 中草药

Objective: To establish a UPLC-MS/MS method for simultaneous determination of 17 bioactive compounds in Shuang-Huang-Lian Oral Liquid. Methods: The separation was performed on an Acquity UPLC HSS T3 C18 column (100 mm×2.1 mm, 1.7 μm) and the flow rate was set at 0.4 mL/min with acetonitrile/methanol (4:1)-0.4% formic acid as the mobile phase. The column temperature was 40℃ and only 7 min was needed for separating the five pairs of isomers. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source with positive and negative modes. Results: All calibration curves had good linearity (r2>0.9901). The precisions and accuracies of the 17 analytes were all satisfied and the recoveries were 95.81%-102.42%. The content ranges of the 17 compounds were 782.68-1034.27μg/mL for neochlorogenic acid, 786.35-1103.77 μg/mL for chlorogenic acid, 898.00-1238.94 μg/mL for cryptochlorogenic acid, 82.85-115.17 μg/mL for caffeic acid, 226.02-461.63 μg/mL for 3,5-dicaffeoylquinic acid, 191.59-404.21 μg/mL for 3,4-dicaffeoylquinic acid, 243.62-298.51 μg/mL for isoforsythiaside, 978.43-1487.37 μg/mL for forsythoside A, 351.97-435.82 μg/mL for forsythin, 41.19-75.65 μg/mL for rutin, 40.28-73.73 μg/mL for baicalein, 10080.54-10820.35 μg/mL for baicalin, 40.88-50.51 μg/mL for scutellarin, 187.73-204.85 μg/mL for wogonoside, 144.92-335.08 μg/mL for oroxylin A-7-O-β-D-glucuronide, 9.30-25.58 μg/mL for wogonin, and 7.51-16.58 μg/mL for oroxylin A, respectively. Conclusion: The developed UPLC-MS/MS method is simple, sensitive, and accurate, and could be applied for simultaneous determination of the 17 components in Shuang-Huang-Lian Oral Liquid.

Contents of Chlorogenic Acid and 3,5-dicaffeoylquinic Acid in Xanthii Fructus Processed by Different Temperature and Time / 中国中医药信息杂志

Objective To study the best processing technology of Xanthii Fructus by determining the contents of chlorogenic acid and 3,5-dicaffeoylquinic acid in which processed by different temperature and time. Methods Sixteen batchs samples of Xanthii Fructus were propressed by stir-frying with sand, and the propressed temperature and time were set at 150-220 ℃ and 0.5-7 minutes. Two phenolic acid components in Xanthii Fructus were simultaneously determined. The column was UPLC Acquity BEH C18 (2.1 mm×100 mm, 1.7 μm). The mobile phase was acetonitrile-0.1% phosphoric acid, gradient elution. The flow rate was 0.25 mL/min, and the detection wavelength was 327 nm. Results The sample with highest contents of chlorogenic acid and 3,5-dicaffeoylquinic acid was the batch processed by stir-frying with sand at 160 ℃ for 7 minute, which was 2.498, 2.004 mg/g, respectively. Conclusion According to the appearance of processed sample and the content of chlorogenic acid and 3,5-dicaffeoylquinic acid, the optimal processing technology of Xanthii Fructus was stir-frying with sand at 160 ℃ for 7 min.

Determination of organic acids and glycosides in different species of Lonicerae Flos / 中草药

Objective: To determine and compare the contents of seven kinds of organic acids (neochlorogenic acid, chlorogenic acid, 4-dicaffeoylquinicacid, caffeic acid, 3, 4-dicaffeoylquinic acid, 3, 5-dicaffeoylquinic acid, and 4, 5-dicaffeoylquinic acid) and secoxyloganin in Lonicerae Flos. Methods: The contents were determined by HPLC. The analysis was carried out with gradient elution on a Phenomenex C18 colunm eluted with methanol and 0.1% phosphoric acid. The detection wavelength was set at 225 nm. Results: Neochlorogenic acid, chlorogenic acid, 4-dicaffeoylquinicacid, caffeic acid, 3, 4-dicaffeoylquinic acid, 3, 5-dicaffeoylquinic acid, and 4, 5-dicaffeoylquinic acid in Lonicerae Flos and secoxyloganin in their linear ranges showed good linear relationship, the average recoveries (n = 9) were 99.41%, 99.05%, 99.01%, 98.71%, 98.79%, 98.82%, 98.89%, and 99.16%, and RSD values were 0.56%, 0.37%, 0.93%, 1.11%, 0.90%, 0.87%, 0.71%, and 1.04%. Conclusion: The method is rapid, simple, accurate, and can be used for quality evaluation of the seven kinds of organic acids in Lonicerae Flos from different germplasm and secoxyloganin.

Chemical constituents in Swertia davidi / 中草药

Objective: To study the chemical constituents from the whole plant of Swertia davidi. Methods: The chemical constituents were isolated and purified with silica gel column chromatography, silica gel chromatography, etc. The structures of the compounds were identified by physicochemical properties and various spectroscopic methods. Results: Sixteen compounds were isolated as uvaol (1), oleanolic acid (2), 5-carboxyl-3, 4-dihydrogen-1H - 2-benzopyran-1-one (3), 3β, 12β-pregnane-16-12-ene-20 (4), swertisin (5), 6-C-β-glucopyranosyl-7-O-methylluteolinidin (6), isoscoparin (7), 6-demethoxy-7-methylcapillarisin (8), 3-acetoxy-28-hydro-xyl-12-eneurs (9), gentiopicroside (10), N-pentacosy-2-carboxy-benzoylamide (11), 4-O-glycosyloxy-2-hydroxy-6-methoxy-acetophenone (12), quercetin-3-O-β-D-xylopyranosyl-(1→2) - β-D-galactopyranoside (13), 8-epikingiside (14), 3, 5-dicaffeoylquinicacid (15), and 2-methylphenyl-1-O-β-D-glucopyranoside (16). Conclusion: All the compounds are isolated from S. davidi for the first time.

Influence of Mg2+ ions on the interaction between 3,5-dicaffeoylquinic acid and HTLV-I integrase / Influencia de los iones Mg+2 sobre la interacción entre el ácido 3,5- dicafeoilquínico y la integrasa del HTLV-I / Influência dos íons Mg2+ sobre a interação entre o ácido 3,5-dicafeoilquínico e a integrase do HTLV-I

Objective. Using molecular simulation, we studied the influence of Mg2+ ions on the binding mode of HTLV-I Integrase (IN) catalytic domain (modeled by homology) with the 3,5- Dicaffeoylquinic Acid (DCQA). HTLV-I Integrase homology model was built using template-like crystallographic data of the IN catalytic domain solved for Avian Sarcoma Virus (VSA, pdb: 1VSD). Materials and methods. In order to analyze the role of Mg2+ in the interaction or coupling between 3,5-DCQA and Integrase, three models were created: i) in the absence of Mg2+ ions, ii) with a Mg2+ ion coordinated at Asp15 and Asp72 and iii) model with two Mg2+ ions coordinated at Asp15-Asp72 and Asp72-Glu108. Coupling force and binding free energy between 3,5-DCQA and HTLV-I IN were assessed in the three models. Results. The lowest docking score and free energy binding were obtained for the second model. Mg2+ ion strongly affected the coupling of the inhibitor 3,5-DCQA with HTLV-I catalytic domain of Integrase, thus revealing a strong interaction in the ligand-protein complex regardless of the ligand-catalytic interaction sites for all three models. Conclusion. Altogether, these results strengthen the hypothesis that the presence of one Mg2+ ion could enhance the interaction in the complex by decreasing free energy, therefore increasing the affinity. Moreover, we propose 3,5-DCQA as an important pharmacophore in the rational design of new antiretroviral drugs.

Objetivo: Usando simulación molecular, estudiamos la influencia de los iones Mg2+ en la interacción del dominio catalítico de la integrasa HTLV-I (IN) (modelado por homología) con el ácido 3,5-Dicafeoilquínico (DCQA). Materiales y métodos. El modelo por homología de la HTLV-I IN fue construido usando como molde la estructura cristalina de la IN del virus del sarcoma aviar (VSA, pdb: 1VSD). Para analizar el rol de los iones Mg2+ en la interacción con el DCQA y la integrasa, tres modelos fueron creados: i) en ausencia de iones Mg2+, ii) con un ion Mg2+ coordinado con Asp15 y Asp72 y iii) con dos iones Mg2+ coordinados con Asp15-Asp72 y Asp72-Glu108. Las fuerzas de interacción y la energía libre de unión entre el DCQA y la HTLV-I IN fueron calculadas en los tres modelos. Resultados. El puntaje más bajo en el docking y la menor energía libre fueron obtenidos para el modelo con un solo ion de Mg2+. El Mg2+ afecta fuertemente el acoplamiento del inhibidor DCQA con el dominio catalítico de la HTLV-I IN, revelando una fuerte interacción entre el complejo ligando-proteína que es independiente del sitio catalítico de los tres modelos usados. Conclusión. Los resultados sugieren que la presencia de un ion de Mg² + podría incrementar la interacción en el complejo debido a la disminución de la energía libre, intensificando así la afinidad. Por lo que, proponemos al DCQA como farmacóforo para el diseño de drogas antiretrovirales.

Objetivo. Usando simulação molecular, estudamos a influência dos íons Mg2+ na interação do domínio catalítico da integrase HTLV-I (IN) (modelado por homologia) com o ácido 3,5-dicafeoilquínico (3,5-diCQA). Materiais e métodos. O modelo de homologia da HTLV-I IN foi construído utilizando como fôrma a estrutura cristalina da IN de vírus do sarcoma aviário (VSA, pdb: 1VSD). Para analisar o papel dos íons Mg2+ na interação com o 3,5-diCQA e a integrase, três modelos foram criados: i) na ausência de íons Mg2+, ii) com um íon Mg2+ coordenado com Asp15 e Asp72 e, iii) com dois íons Mg2+ coordenados com Asp15-Asp72 e Asp72-Glu108. As forças de interação e a energia livre de ligação entre o 3,5-diCQA e a HTLV-I IN foram calculadas nos três modelos. Resultados. A pontuação mais baixa no docking e a menor energia livre foram obtidas para o modelo com um único íon de Mg2+. O Mg2+ afeta fortemente o acoplamento do inibidor 3,5-diCQA com o domínio catalítico da HTLV-I IN, revelando uma forte interação entre o complexo ligando-proteína que é independente do sítio catalítico dos três modelos utilizados. Conclusão. Os resultados sugerem que a presença de um íon Mg2+ poderia aumentar a interação no complexo devido à diminuição da energia livre, aumentando assim a afinidade. Assim, propomos ao 3,5-diCQA como farmacóforo para a fabricação de medicamentos antirretrovirais.

Identification of the antimicrobial substances produced by Solanum palinacanthum (Solanaceae)

To find out natural antimicrobial agents as alternative in therapeutics and to preserve food, the methanol extract of Solanum palinacanthum aerial parts was submitted to purification steps guided by antibacterial and antifungal assays. As a consequence, the flavonoid rutin and 3,5-dicaffeoylquinic acid were isolated by column chromatographyand high performance liquid chromatography, and identified by mass and nuclear magnetic resonance spectrometry. Minimal inhibitory concentrations (MIC) of the quinic acid derivative against Aeromonas hydrophila, Bacillus subtilis, Staphylococcus aureus and the fungus Aspergillus ochraceus were 250, 1000, 1000 and > 568µg/mL, respectively. Against the same microorganisms, MIC for rutin were 1000, > 1000, > 1000 and 35µg/mL, respectively. Rutinwas very promising for A. ochraceus control, since its MIC against such fungus was close to the one observed for benzalkonium chloride, which is used as a fungicide in Brazil.

Com vistas a descobrir antimicrobianos de origem natural para uso terapêutico ou para a preservação de alimentos, o extrato metanólico das partes aéreas de Solanum palinacanthum foi submetido a fracionamentos direcionados por testes para avaliar a atividade antibacteriana e antifúngica. Em decorrência, o flavonóide rutina e o ácido 3,5-dicafeoilquínico foram isolados por cromatografia em coluna e por cromatografia líquida de alta eficiência, para serem identificados por espectrometria de massas e de ressonância magnética nuclear. As concentrações inibitórias mínimas (CIM) do derivado do ácido cafeico contra Aeromonas hydrophila, Bacillus subtilis, Staphylococcus aureus e o fungo Aspergillus ochraceus foram 250, 1000, 1000 e > 568µg/mL, respectivamente. Contra os mesmos organismos, os valores de CIM para a rutina foram 1000, > 1000, > 1000 e 35µg/mL, respectivamente. A rutina mostrou-se muito promissora para o controle de A. ochraceus, pois seu valor de CIM contra tal fungo foi bem próximo ao observado para o cloreto de benzalcônio, que é empregado como fungicida no Brasil.

Determination of Six Phendic Acids in Mailuoning Injection by RP-HPLC / 中国中医药信息杂志

Objective To establish a method for determination the contents of six phendic acids in Mailuoning injection by HPLC at same time. Methods Separation was performed on a Kromasil-C18 (4.6 mm? 250 mm, 5 ?m) column with mobile phase consisting of 0.1% phosphoric acid-water and Acetonitrile with gradient elute at the flow rate of 1 mL/min. The UV detection wavelength was set at 327 nm and the column temperature was set at 25 ℃. The sample size was 20 ?L. Results The standard cures of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, 3,5-Dicaffeoylquinic acid, 3,4- Dicaffeoylquinic acid were linear within the contentration range of 0.010 90～0.544 8 ?g (r=0.999 9), 0.024 06～1.203 2 ?g (r =1), 0.011 06～0.552 8 ?g (r =1), 0.015 94～0.796 8 ?g (r =1), 0.009 104～0.455 2 ?g (r =1), 0.010 37～0.518 4 ?g (r=1) respectively. And the recovery rates of them were 101.06%, 100.22%, 101.54%, 99.42%, 100.21%, 101.65% respectively. Conclusion The method appeared to be simple, reliable, accurate and can be used to determine the contents of six phendic acids in Mailuoning injection.