Regulatory effect of eight Chinese herbal medicines on glucose and lipid metabolism and their potential active components based on HCBP6 target / 中国中药杂志

With the improvement of living standards and changes in working style, the prevalence of abnormal glucose and lipid metabolism in humans is increasing in modern society. Clinically, the related indicators are often improved by changing the lifestyle and/or taking hypoglycemic and lipid-lowering drugs, but there are no therapeutic drugs for disorders of glucose and lipid metabolism at present. Hepatitis C virus core protein binding protein 6(HCBP6) is a newly discovered target that can regulate triglyceride and cholesterol content according to level oscillations in the body, thereby regulating abnormal glucose and lipid metabolism. Relevant studies have shown that ginsenoside Rh_2 can significantly up-regulate the expression of HCBP6, but there are few studies on the effect of Chinese herbal medicines on HCBP6. Moreover, the three-dimensional structural information of HCBP6 has not been determined and the discovery of potential active components acting on HCBP6 is not rapidly advanced. Therefore, the total saponins of eight Chinese herbal medicines commonly used to regulate abnormal glucose and lipid metabolism were selected as the research objects to observe their effect on the expression of HCBP6. Then, the three-dimensional structure of HCBP6 was predicted, followed by molecular docking with saponins in eight Chinese herbal medicines to quickly find potential active components. The results showed that all total saponins tended to up-regulate HCBP6 mRNA and protein expression, where gypenosides showed the optimum effect on up-regulating HCBP6 mRNA and ginsenosides showed the optimum effect on up-regulating HCBP6 protein expression. Reliable protein structures were obtained after the prediction of protein structures using the Robetta website and the evaluation of the predicted structures by SAVES. The saponins from the website and literature were also collected and docked with the predicted protein, and the saponin components were found to have good binding activity to the HCBP6 protein. The results of the study are expected to provide ideas and methods for the discovery of new drugs from Chinese herbal medicines to regulate glucose and lipid metabolism.

Quality evaluation methods of Poria based on yield and content of alkali extracted polysaccharide / 中国中药杂志

To improve the quality control methods of Poria and develop and utilize its resources fully, alkaline extraction was used in this study to determine the yield and content of alkali-soluble polysaccharides of Poria. The alkali-soluble extracts of Poria were obtained according to the optimum extraction conditions on the basis of single-factor test, and 30 batches of samples were determined. The structure and chemical composition of the alkali-soluble extracts was characterized by high-performance gel permeation chromatography(HPGPC), Fourier transform infrared spectrometry(FT-IR), nuclear magnetic resonance(NMR) spectroscopy and high-performance liquid chromatography(HPLC) with 1-phenyl-3-methyl-5-pyrazolone(PMP-HPLC). The results showed that the content of the alkali-soluble extracts was in the range of 46.98%-73.86%. The main component was β-(1→3)-glucan, and its molecular mass was about 1.093×10~5. Further, the content of alkali-soluble polysaccharides of Poria was measured by UV-Vis spectrophotometry and HPLC coupled with the evaporative light scattering detector(HPLC-ELSD), and 30 batches of samples were measured. The results indicated that the content of alkali-soluble polysaccharides determined by UV-Vis spectrophotometry was in the range of 73.70%-92.57%, and the content of samples from Hubei province was slightly higher than that from Yunnan province, Anhui province and Hunan province. The content of alkali-soluble polysaccharides determined by HPLC-ELSD was in the range of 51.42%-76.69%, and the samples from Hunan province had slightly higher content than that from the other three provinces. The content determined by UV-Vis spectrophotometry was higher than that by HPLC-ELSD. However, the content determined by HPLC-ELSD was close to that of alkali-soluble extract, which could accurately characterize the content of alkali-soluble polysaccharides in Poria, and the method was simple and repeatable. Therefore, it is recommended that the quantitative analysis method for alkali-soluble extract and alkali-soluble polysaccharides by HPLC-ELSD be used in the quality standards of Poria in Chinese Pharmacopeia.

Comparison of chemical components between aerial and underground parts of Coptis chinensis based on UPLC-Q-TOF-MS~E technology / 中国中药杂志

The ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS~E) technology was employed to compare the chemical components between the aerial and underground parts of Coptis chinensis samples from different batches. According to the retention time, molecular ion peak, and LC-MS~E fragment information of the reference substances and available literature, we identified a total of 40 components. Thirty-three and 31 compounds were respectively identified in the underground part(taproots) and the aerial part(stems and leaves) of C. chinensis. Among them, 24 compounds, including alkaloids(e.g., berberine and jatrorrhizine) and phenolic acids(e.g., chlorogenic acid, quinic acid, and tanshinol), were common in the two parts. In addition, differential components were also identified, such as magnoline glucoside in the underground part and(±) lariciresionol-4-β-D-glucopyranoside in the aerial part. The analysis of fragmentation pathways based on spectra of reference substances indicated the differences among samples of different batches. Furthermore, we performed the principal component analysis(PCA) for the peak areas of C. chinensis in different batches. The results showed that the underground part and the aerial part were clearly clustered into two groups, indicating that the chemical components contained in the two parts were different. Furthermore, the results of partial least squares discriminant analysis(PLS-DA) identified 31 differential compounds(VIP value>1) between the underground part and the aerial part, mainly including alkaloids, phenolic acids, lignans, and flavonoids. This study proves that C. chinensis possesses great development potential with multiple available compounds in stems and leaves. Moreover, it sheds light on for the development and utilization of non-medicinal organs of C. chinensis and other Chinese medicinal herbs.

Qualitative and quantitative analysis of nucleosides and nucleobases in Phylloporia ribis by UPLC-Q-TOF-MS and HPLC / 中国中药杂志

The dried fruit body of Phylloporia ribis(Hymenochaetaceae), which prefers to live on the stumps of Lonicera japonica(Caprifoliaceae), has a variety of activities, whereas its pharmacodynamic material basis is not completely clear and there are few reports on its quality control and evaluation. In this study, an UPLC-Q-TOF-MS method was used to analyze the nucleosides and nucleobases in P. ribis and a HPLC method was established for simultaneous determination of 10 nucleosides and nucleobases. MS and MS/MS data were acquired in positive ion mode. Based on the data comparison of the sample and the reference substance, the literature data and the compound databases of ChemSpider and PubChem, 18 nucleosides and nucleobases were identified qualitatively from the water extract of P. ribis for the first time. After optimization, the HPLC was performed using a Welch Ultimate AQ C_(18) column(4.6 mm×250 mm, 5 μm) by gradient elution with acetonitrile and water as mobile phase, the flow rate of 1.0 mL·min~(-1), the detection wavelength of 260 nm, and the column temperature of 30 ℃. Through the investigation of the extraction method, solvent and time, it was determined that the test solution should be obtained by cold water extraction for 18 h. At the present HPLC conditions, 10 components of uracil, cytidine, hypoxanthine, uridine, thymine, inosine, guanosine, 2'-deoxyinosine, 2'-deoxyguanosine and thymidine could be well separated(R > 1.5) and showed good linearity(r > 0.999 9) in the concentration ranges of 0.247-24.7, 0.283-28.3, 0.273-27.3, 0.256-25.6, 0.257-25.7, 0.318-31.8, 0.245-24.5, 0.267-26.7, 0.250-25.0 and 0.267-26.7 mg·L~(-1), respectively. The average reco-veries of 10 components were 95.78%-104.5%, and the RSDs were 2.2%-5.2%(n=6). The contents of 10 nucleosides and nucleobases in different samples of P. ribis varied greatly, which were 0.021-0.122, 0.004-0.029, 0.014-0.226, 0.009-0.442, 0.003-0.014, 0.002-0.146, 0.007-0.098, 0-0.054, 0.005-0.069, 0.004-0.081 and 0.072-1.28 mg·g~(-1) for uracil, cytidine, hypoxanthine, uridine, thymine, inosine, guanosine, 2'-deoxyinosine, 2'-deoxyguanosine, thymidine and total 10 components, respectively. These results demonstrated that the components had significant differences in the internal quality, and good quality control was needed to ensure the medical efficacy. This study provides a scientific basis for the discovery of pharmacodynamic ingredients, quality control and evaluation of P. ribis.

Research progress of pesticide residues in Chrysanthemum / 中国中药杂志

Chrysanthemum is widely used as a type of edible flower and also considered as the important materials of many beverages in China. Due to the occurrence of diseases and pests, and the lack of regulations for species, frequency, dose of pesticides in Chrysanthemum, pesticides have become one of the main pollutants in Chrysanthemum. The pesticide residues in Chrysanthemum were detected frequently and worth noting. This paper focused on the types of pesticides, pesticide residue detection techniques, and risk assessment methods for Chrysanthemums on the basis of relevant literatures. The pesticide residues of traditional Chinese medicine are mainly organochlorines, organophosphorus and pyrethroids, and the detection techniques include gas chromatography(GC), liquid chromatography(LC) or both combined with mass spectrometry(MS). With the increasing use of traditional Chinese medicine, Chrysanthemum is widely circulated in the market. Therefore, it is important to understand the current situation of pesticide residues in different varieties of Chrysanthemum, so as to provide theoretical reference for the control of quality and safety of Chrysanthemum and the formulation of the maximum residue limit.

Scientificity of traditional commodity grade of Chinese medicinal materials--taking Lonicerae Japonicae Flos as example / 中国中药杂志

Taking Lonicerae Japonicae Flos as an example, the method of "expert consensus of different regions" was used to screen the representative samples and evaluate their commodity grades. The correlation analysis, hierarchical cluster analysis and partial least squares discriminant analysis(PLS-DA) of "commodity grade-appearance characteristic-component content" were carried out to reveal the scientificity of traditional commodity grade of Chinese medicinal material. By referring to the existing literature and the grade investigation from the sample collection regions, 78 "initial grade" samples were screened out from 118 collected samples. Authoritative experts from four regions(n=4) including Linyi(Shangdong province), Bozhou(Anhui province), Anguo(Hebei province) and Beijing were organized to evaluate their commodity grades, separately. Based on the grade consistency rate(R_i≥70%), 69 "local grade" samples were screened out from the "initial grade" samples. Based on the average grade consistency rate ■ "authoritative grade" samples were screened out from the "local grade" samples, including15 first-grade samples, 9 second-grade samples, 11 third-grade samples and 17 fourth-grade samples. For these "authoritative grade" samples, the main appea-rance characteristics were quantified and the contents of 13 components were determined by ultra performance liquid chromatography(UPLC). Furthermore, the total contents of 6 phenolic acids, 4 flavonoids and 3 iridoids were calculated, respectively. The results of correlation analysis showed that 4 appearance characteristics indices were correlated with the commodity grades: color, rate of yellow bars(including blooming flowers), rate of black heads(including black bars), and rate of stems and leaves(including bud debris). Five component content indices were correlated with the commodity grades: chlorogenic acid, isochlorogenic acid C, sweroside, loganin and the total contents of six phenolic acids. Furthermore, chlorogenic acid, loganin and the total contents of six phenolic acids showed significantly negative correlation with the main appearance characteristics, indicating that the appearance characteristics of Lonicerae Japonicae Flos can reflect its internal quality, and these 3 indices can be used as quality markers(Q-markers). The results of hierarchical cluster analysis showed that the samples of four grades were classified into four categories, and the samples with the same grades and the same categories accounted for 80.8% of the total samples, while the samples with the different grades were obviously classified into different categories. The results of PLS-DA analysis showed that the samples of different grades showed obvious intragroup aggregation and intergroup dispersion. The above results indicated that it was feasible to evaluate the traditional commodity grade of Lonicerae Japonicae Flos by the method of "expert consensus of different regions". For the evaluation of traditional commodity grade of Chinese medicinal material, the samples should be representative, expert conclusions should have enough consensuses, and grade determination should be authoritative. As the crystallization of clinical experience, traditional commodity grade can scientifically reflect the internal quality of Chinese medicinal material.

Establishment of QAMS method with dehydroandrographolide as internal reference substance for quality control of Andrographis Herba / 中国中药杂志

To solve the problems of the poor resolution of chromatographic separation,the weak durability of the relative correction factors,and the low accuracy of content determination results in the quantitative analysis of multi-components by single-marker( QAMS) method with andrographolide as the internal reference substance in the existing research of Andrographis Herba,a new QAMS method using dehydroandrographolide as the internal reference substance was established for the first time in this study. This new method can be used to simultaneously determine four diterpene lactones,including andrographolide( A),neoandrographolide( B),14-deoxyandrographolide( C),and dehydroandrographolide( S) through the optimization of chromatographic conditions and systematic investigation of methodology. At the present HPLC chromatographic conditions,four components could be well separated( R > 1. 5),and the methodology validations could satisfy the requirement of quantitative analysis. The relative correction factors( RCFs) of fA/S,fB/S,fC/S were determined as 0. 65,0. 54,0. 78,respectively. The relative standard deviations( RSDs) of their RCFs ranged between 1. 3%-5. 1%,0. 25%-0. 33%,0. 070%-0. 15%,0. 070%-0. 22%,respectively with three brands of HPLC instruments,five brands of C18 column,different flow rates( 0. 9,1. 0,1. 1 m L·min~(-1)),and different column temperatures( 25,30,35 ℃),indicating good durability of the RCFs. The relative retention value( RRV) method was used to locate the chromatographic peak of the components to be determined.The RRVs of rA/S,rB/S,and rC/Swere 0. 44,0. 86,0. 97,respectively. The RSDs of the RRVs ranged between 0. 030%-1. 6% with different HPLC instruments and columns,showing accurate peak location. The present QAMS method and the external standard method( ESM)were both used to determine the contents of four diterpene lactones from Andrographis Herba( 6 batches of medicinal materials and 18 batches of cut crude drugs). The relative errors of the determined content results between two methods were less than 2. 0%. It demonstrated that there was no significant difference in content results between these two methods,indicating good accuracy of the present QAMS method. Therefore,in this study,an accurate and highly durable QAMS method using dehydroandrographolide as the internal reference substance was established for simultaneous determination of four diterpene lactones. This method could be used to effectively control the quality of Andrographis Herba and provide technical basis for the formulation of traditional Chinese medicine industry standard and improvement of the Chinese Pharmacopoeia standard of Andrographis Herba.

Analysis of standard decoction of salt-processed Psoraleae Fructus / 中国中药杂志

The traditional Chinese medicine standard decoction is prepared on the basis of the theory of traditional Chinese medicine and clinical application. With reference to the modern extraction method,the single decoction of traditional Chinese medicine is prepared by the standardized process,and the establishment of its quality standards is conducive to standardizing clinical medication. This research is to set an evaluation standard for the quality of salt-processed Psoraleae Fructus standard decoction. Twelve batches of salt-processed Psoraleae Fructus standard decoctions were prepared. The contents of psoralen and isopsoralen were determined,the transfer and extract rates were calculated,and the pH value was measured; HPLC fingerprint method was established for analysis. The results of the 12 batches of samples revealed that the transfer rates of psoralen and isopsoralen were 17. 10%-26. 40%,14. 70%-22. 70%,respectively; the extract rate was between 14. 7%-27. 0%,and the pH value was between 5. 4-6. 9. Moreover,7 common chromatographic peaks were determined based on fingerprint by using similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine( 2012 A).The similarities of the 12 batches of samples were analyzed and compared,and the results showed that the similarities were all higher than0. 9. In this study,the preparation method for salt-processed Psoraleae Fructus decoction was standard,with high similarities in fingerprint. This study build a convenient and reliable method of comprehensive quality evaluation,with a high precision,stability and repeatability,which can provide a reference for the quality control of salt-processed Psoraleae Fructus dispensing granules.

Application of Mycobacterium marinum infected zebrafish for evaluation of anti-tuberculosis drugs / 药学学报

Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium. tuberculosis. In recent years, with the emergence of drug-resistant forms, the development of new anti-tuberculosis drugs is urgently needed. In this study, we used Mycobacterium marinum (M. marinum), which is highly similar to M. tuberculosis, to establish a M. marinum infected-zebrafish model and quantitative PCR (qPCR) method for bacterial count analysis. The results showed that injecting M. marinum into the yolk sac is an efficient and convenient way to infect zebrafish embryos. By counting the survival rate of infected zebrafish and the number of bacteria in zebrafish by Ziehl-Neelsen staining, we analyzed the efficacy of isoniazid and rifampicin as anti-tuberculosis drugs and the synergistic effect of drugs. The results suggested that three evaluation methods exhibit good consistency. This study demonstrated that zebrafish-M. marinum infection model combined with qPCR analysis is a simple and efficient method for in vivo screening and evaluation of anti-tuberculosis drugs. Animal experiments were carried out in accordance with the provisions for animal ethics in the Regulations on Laboratory Animals of Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences.

Quantitative analysis of nine ginsenosides in red ginseng using QAMS method based on high-resolution and reference chromatography of red ginseng / 中国中药杂志

Using reversed-phase high performance liquid chromatography, nine ginsenosides were simultaneously separated on an UltimateC₁₈ column with high-resolution and high purity of each chromatographic peak. Adopting the QAMS quality evaluation model for traditional Chinese medicines, ginsenoside Rb₁ was used as the internal reference substance, and the relative correction factors (RCFs) and the relative retention values (RTRs) of ginsenosides Rg₁, Re, Rf, Rb₁, Rc, Rb₂, Rb₃, Rd and 20 (S)-ginsenoside Rg₃ to ginsenoside Rb₁ were calculated individually. Through a series of methodology evaluations, and positioned by the red ginseng reference chromatograph and RTVs, nine ginsenosides in red ginseng were simultaneously assayed only by quantitative determined ginsenoside Rb₁.

Exploration on feasibility of introducing bioassay method into quality evaluation of Chinese herbal medicines by studying on the correlation between antioxidant activity of Prunella vulgaris and its total phenolic acids content for example / 中国中药杂志

This paper aims to investigate the correlation between the antioxidant activity of Prunella vulgaris and its total phenolic acids content by measuring the antioxidant activity of different sources and different organs of P. vulgaris and the total contents of protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid in these samples. Using the 50% methanol extract of P. vulgaris samples as the research object, DPPH method and HPLC method were used respectively to determine the antioxidant activities and the total contents of the above-mentioned five analytes in P. vulgaris samples. 0.5 mL of 50% methanol extract of P. vulgaris reacts with 0.1 mmol•L⁻¹ DPPH ethanol solution for 60 min, then the absorbance of the reaction solution was measured at 517 nm, scavenging rate and IC₅₀ values were calculated by the absorbance and the sample concentration for evaluating the antioxidant activity. HPLC analysis was made on a C₁₈ Epic column, with acetonitrile-0.1% formic acid aqueous solution as mobile phase (gradient elution), and the detection wavelength was set at 280 nm. The correlation between the antioxidant capacity of different habitats and different organs of P. vulgaris and the total contents of five kinds of phenolic acids was analyzed by partial least squares method. The reaction dose-response range of 50% methanol extract of P. vulgaris with 0.1 mmol•L⁻¹ DPPH ethanol solution was 0.300-1.65 g•L⁻¹. When the quantities of potocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid were respectively in 0.007 84-0.980, 0.011 5-1.44, 0.008 64-1.08, 0.080 0-1.00 and 0.079 8-0.998 μg range, their quantities were in good linear relationship with the corresponding peak areas. The average recovery of 5 components were 97.76%, 96.88%, 100.3%, 102.1%, 104.5%, with RSD of 1.8%, 1.6%, 1.7%, 1.6% and 1.7%, respectively. In a certain range of crude drug quantity, the antioxidant activity of each organ of P. vulgaris and total phenolic acids content inside has a good linear correlation. Therefore, in certain quality range of crude drug, DPPH bioassay combined with HPLC content determination can be used for the quality control of P. vulgaris, as is a new method for the quality control of P. vulgaris.

Integrative research of QAMS model used in assay of flavonoids in Epimedii Folium, its processed products and finished Chinese medicines / 中国中药杂志

Through a series of methodology investigations, we established a new method for simultaneous analysis of epimedins A, B, C, icariin and baohuoside I in Epimedii Folium by using high performance liquid chromatography (HPLC). Meanwhile, using Icariin as an internal reference substance to establish the relative correct factors and relative retention values of Epimedins A, B, C and Baohuoside I to Icariin, then using the quantitative analysis of multi-components by single-marker (QAMS) model, the five analytes can be quantitatively determined in Epimedii Folium and its processed products as well as Kanggu Zengsheng capsule only by measuring the content of icariin in the corresponding samples. All these analysis are completed in the same chromatorgraphic conditions. This paper played the part of demonstration role in the popularization and application of QAMS method established in a single herb to the proprietary Chinese medicines containing this herb.

Study on antioxidant chemical constituents of Lonicera japonica leaves / 中国中药杂志

Guided by the antioxidant activity, the EtOAc-soluble and n-butanol-soluble fractions of the 50% methanol extract of Lonicera japonica leaves were isolated and purified by various chromatorgraphic methods, and the structures were identified by spectral analysis and comparison to the data reported in literature. As a result, nine compounds were obtained and identified as 5-O-caffeoylquinicacid (1), chlorogenicacid (2), 4-O-caffeoylquinicacid (3), luteolin-7-O-[α-L-arabinopyranosyl-(1 --> 6)] -β-D-glucopyranoside (4), luteoloside (5), 3,4-di-O-caffeoylquinic acid (6), 3,5-di-O-caffeoylquinic acid (7), 4, 5-di-O-caffeoylquinic acid (8) and luteolin (9). The antioxidant activity of the nine compounds were determined by using DPPH free radical scavenging method, and ascorbic acid was used as a positive control. Their antioxidant activities from high to low were 5 > 9 > 2 > 8 > 7 > 6 > 1 > 3 > 4. Among them, luteoloside (5) had the strongest antioxidant activity with an IC50 of 0.018 18 g x L(-1), and luteolin (IC50 0.023 6 g x L(-1)) and chlorogenicacid (IC50 0.035 17 g x L(-1)) ranks No. 2 and 3. Furthermore, the antioxidant activity of luteoloside and luteolin were stronger than that of ascorbic acid (IC50 0.027 54 g x L(-1)). These results gave a basis for the further study and utilization of L. japonica leaves.

Establishment of simultaneous quantitative model of five alkaloids from Corydalis Rhizoma by near-infrared spectrometry / 中国中药杂志

This paper established a near-infrared spectroscopy quantitative model for simultaneous quantitative analysis of coptisine hydrochloride, dehydrocorydaline, tetrahydropalmatine, corydaline and glaucine in Corydalis Rhizoma. Firstly, the chemical values of the five components in Corydalis Rhizoma were determined by the reversed-phase high performance liquid chromatography (RP-HPLC) with UV detection. Then, the quantitative calibration model was established and optimized by fourier transformation near-infrared spectroscopy (NIRS) combined with partial least square (PLS) regression. The calibration model was evaluated by correlation coefficient (r), the root-mean-square error of calibration (RMSEC) and the root mean square of cross-validation (RMSECV) of the calibration model, as well as the correlation coefficient (r) and the root mean square of prediction (RMSEP) of prediction model. For the quantitative calibration model, the r, RMSEC and RMSECV of coptisine hydrochloride, dehydrocorydaline, tetrahydropalmatine, corydaline and glaucine were 0.941 0, 0.972 7, 0.964 3, 0.978 1, 0.979 9; 0.006 7, 0.003 5, 0.005 9, 0.002 8, 0.005 9; and 0.015, 0.011, 0.020, 0.010 and 0.022, respectively. For the prediction model, the r and RMSEP of the five components were 0.916 6, 0.942 9, 0.943 6, 0.916 7, 0.914 5; and 0.009, 0.006 6, 0.007 5, 0.006 9 and 0.011, respectively. The established near-infrared spectroscopy quantitative model is relatively stable, accurate and reliable for the simultaneous quantitative analysis of the five alkaloids, and is expected to be used for the rapid determination of the five components in crude drug of Corydalis Rhizoma.

Construction of cDNA infectious clones of EV71 highly-pathogenic and cell-culture-adapted strains / 病毒学报

The highly-pathogenic EV71 strain is the primary cause of mortality in hand-foot-and-mouth disease (HFMD) associated with neurological symptoms, for which no clinically effective drugs or vaccines exist. This study aimed to construct infectious cDNA clones of the EV71 highly-pathogenic strain and the cell-culture adapted strain using a reverse genetics approach. The genomic RNAs of EV71 parent strains were used as the templates for RT-PCR amplification, and then the PCR products that overlapped the full-length genome were connected into the pBR322 vector to produce infectious clones of pEV71 (HP) and pEV71 (CCA), respectively. The results showed that the HP strain propagated much more quickly than the CCA strain. The rescued viruses derived from the infectious clones not only maintained their consistency with their parent strains in terms of genomic sequences, but also retained their respective biological phenotypes. This research will contribute to our understanding of EV71 pathogenesis and the development of novel vaccines against HFMD.

Simultaneous determination of seven constituents in Euodiae Fructus and two related species by HPLC / 中国中药杂志

This study is to develop a HPLC method for quality evaluation of Euodiae Fructus and related species by simultaneous determination limonin, indole alkaloids (14-fomyldihydroxyrutaecarpine, evodiamine, rutaecarpine), and quinolone alkaloids [1-methyl-2-undecyl-4 (1H)-quinolone, evocarpine, dihydroevocarpine] in the fruits of five Evodia species. Samples were analyzed on a YMC C18 column (4.6 mm x 250 mm, 5 microm) eluted with mobile phases of acetonitrile (A), tetrahydrofuran (B), and a buffer solution of 5 mmol x L(-1) ammonium acetate (pH 3.8) (C) in a linear gradient mode. The column temperature was 30 degrees C and the flow rate was 1.0 mL x min(-1). The PDA detector wavelengths were set at 220 and 250 nm. The seven compounds were well separated and showed good linearity (r = 0.999 9) within the concentration ranges tested. The mean recoveries were between 96.7%-102.4% (RSD 1.4%-3.1%). Through the validation, the method was proved to be accurate and repeatable. All the seven constituents were detected in the fruits of five species, but the contents of them varied widely in different samples. The total contents of seven constituents in 16 batches of Euodiae Fructus were 9.46-69.9 mg x g(-1), and the mean content was 28.2 mg x g(-1). The total content of seven constituents in E. compacta and E. fargesii was 25.8, 7.69 mg x g(-1), respectively.

Simultaneous determination of five gingerols in raw and processed ginger by HPLC / 中国药学杂志

OBJECTIVE: To develop an HPLC method to determine and compare the contents of zingerone, 6-gingerol, 8-gin-gerol, 6-shogaol, 10-gingerol in fresh, dried, roasted, and charry ginger, and to discuss the change regularity of the five components during the processing of ginger. METHODS: The separation was performed on a Kromasil-C18 column(4.6 mm × 250 mm, 5 μm) with gradient elution (0 - 10 min, B: 40%; 10-40 min, B: 40% → 90%; 40-45 min, B: 90% → 100%; 45-50 min, B: 100% → 40%) of mobile phase composed of acetonitrile (B) and 0.1% aqueous acetic acid (A). The column temperature was set at 30°C. The flow rate was 1.0 mL · min-1 and the injection volume was 10 μL. The detection wavelength was set at 275 nm. RESULTS: The calibration curve was linear over the concentration of 0.625 - 20 μg · mL-1 for zingerone (r = 0.9991); 6.25 - 200 μg · mL-1 for 6-gingerol (r =0.9997); 1.25-40 μg · mL-1 for 8-gingerol (r =0.9999); 1.25-40 μg · mL-1 for 6-shogaol (r =0.9991); 1.875-60 μg · mL-1 for 10-gingerol (r = 0.9994). The average recoveries of the five components were 101.5%, 100.9%, 104.7%, 102.1% and 105.5% and the RSDs were 1.26%, 1.60%, 3.35%, 2.98% and 3.05%, respectively. The contents of 6-gingerol, 8-gingerol, and 10-gingerol gradually decreased from fresh ginger to roasted ginger, dried ones and charry ginger, while the contents of 6-shogaol increased in the same sequence. Little zingerone emerged in the preparation of roasted ginger, but its content increased in charry ginger. CONCLUSION: The method is reproducible, accurate, and can be used for the quality control of raw and processed ginger. In the processing of ginger, the five components have changes both in quality and quantity. Copyright 2012 by the Chinese Pharmaceutical Association.

Dynamic change of paeoniflorin content in root of Paeonia lactiflora growing at Duolun County, Inner Mongolia / 中国中药杂志

<p><b>UNLABELLED</b>To explore the dynamic change of paeoniflorin content in root of Paeonia lactiflora.</p><p><b>METHOD</b>The samples were collected and paeoniflorin content was determined by HPLC.</p><p><b>RESULT</b>The results showed that the average content of paeoniflorin was the lowest at 3.05% in the roots collected in May, and during the fruit time it was rather high in August at 4.72%, and in September at 4.58%. The average content showed no significant differences during June to October.</p><p><b>CONCLUSION</b>Wild plant of P. lactiflora might be collected for medical use during blooming stage until wilting stage.</p>

Determination of chyodeoxycholic acid in Bile Arisaema by HPLC-ELSD / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an HPLC method for the determination of hyodeoxycholic acid in Bile Arisaema.</p><p><b>METHOD</b>The methanol extracts were separated on a Diamonsil ODS column eluted at 35 degrees C with a mixture of methanol-water-glacial acetic acid (75:25:0.01) at flow-rate 1.0 mL x min(-1). The drift-tube temperature of ELSD was 85 degrees C, and the flow rate of air was 3.0 L x min(-1).</p><p><b>RESULT</b>Hyodeoxycholic acid in methanol extract was well separated, the relationship of logarithms of injection amount and peak area was linear (r = 0.9998) over the range of 2.4-24 microg. The average recovery was 97.1% (RSD 2.9%, n=5).</p><p><b>CONCLUSION</b>The method could be used for quality control of Bile Arisaema.</p>

Determination of HPLC fingerprint of Qingfengtong capsule / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a fingerprint analysis method of Qingfengtong capsule by HPLC.</p><p><b>METHOD</b>The samples were extracted with 70% ethanol in an ultrasonic bath. The extracts were analyzed at 35 degrees C on a Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) with 0.1 mol x L(-1) potassium dihydrogen phosphate water-solution as mobile phase A, and methanol as mobile phase B. The analysis followed a linear gradient elution program. Initial condition of the mobile phase was 10% B for 2 minutes; then changed to 90% B in 40 minutes. The flow rate was kept at 1.0 ml x min(-1) and the detector wavelength was 262 nm.</p><p><b>RESULT</b>The main peaks in fingerprint chromatogram of Qingfengtong capsules were separated fairly well. The results of method validation meet the requirements for the fingerprints.</p><p><b>CONCLUSION</b>The established method can be used for the quality control of Qingfengtong capsules.</p>