Application of an effect-constituents index for the quality evaluation of the traditional Chinese medicine rhubarb / 药学学报

To explore the application of an effect-constituents index (ECI) for the quality evaluation of rhubarb, we carried out the simultaneous determination of 12 chemical components by ultra-performance liquid chromatography and used the ICR mouse constipation model to determine the diarrhea biopotency of these 12 components. With the diarrhea biopotency of sennoside A as a reference, the diarrhea biopotency weight coefficient of each chemical component was obtained. A multi-component chemical quantitative analysis combined with the biopotency weight coefficients for rhubarb was developed, named the diarrhea ECI. Animal experiment ethics requirements were approved by the Animal Experimental Ethics Committee of the 302 Hospital of the People's Liberation Army (Grant Number: IACUC-2015-012). The results showed that there were significant differences in the content of the 12 chemical components in different batches of processed products of rhubarb. Especially worthy of attention was the content of aloe-emodin-8-O-β-D-glucoside in sample Rh03, nearly 40-fold higher than that in Rh07 (4.79 vs 0.12 mg·g-1), and the content of rhein-8-O-β-D-glucoside in sample Rh03, nearly 45 times higher than that in Rh07 (3.56 vs 0.08 mg·g-1). The actual measured diarrhea biopotencies of the 12 chemical components ranged from 61.65 ± 4.28 to 233.84 ± 5.58 U·mg-1. The calculated diarrhea effect-constituents indices of 16 rhubarb samples ranged from 1.07 (Rh15) to 19.38 (Rh03), and the actual measured diarrhea biopotencies of 16 rhubarb samples based the ICR mouse constipation model ranged from 23.84 U·g-1 (Rh16) to 310.94 U·g-1 (Rh05). The correlation between the diarrhea ECIs and the actual measured diarrhea biopotencies of 16 rhubarb samples was good (r = 0.969 5), suggesting that the diarrhea effect-constituents indices may be the most suitable for evaluating the quality of different rhubarbs with regard to diarrhea.

Research progress on Q-biomarker of Chinese materia medica / 中草药

Quality marker (Q-marker) is one of the new directions in the research of quality control of Chinese materia medica. The Q-marker can be either quality chemomarker (Q-chemomarker) or quality biomarker (Q-biomarker). The establishment of a good Q-biomarker can improve the linkage of Chinese medicine quality control methods and standards with clinical safety and effectiveness, at the same time considering the operability of quality control. At present, the FDA of China and the United States have adopted biological evaluation methods as an important part of quality control of Chinese medicines and botanicals medicines. Many companies and researchers are also actively exploring and developing methods for the evaluation of quality of Chinese materia medica. This paper further enlarges and expands the methods and indicators of biological evaluation of Chinese materia medica, and summarizes and analyzes relevant research examples from the perspective of Q-biomarker. We hope that it would promote the richness and development of the theoretical system of Chinese medicine quality markers, so as to provide a reference to solve the practical problems for the quality control and evaluation of Chinese materia medica.

Enhancement Study of Macrophage Phagocytic Activity and the Quality Evaluation of Dioscorea opposita Thunb. Based on the Analysis of High Content Screening / 中国药学杂志

OBJECTIVE: To compare the differences of macrophages phagocytic activities between Dioscorea opposita Thunb. cv. Tiegun(abbreviated as TG) and Dioscorea opposita Thunb. but not Tiegun(abbreviated as NTG). METHODS: CCK-8 assay was used to test the cytotoxicity. On the basis of non-toxic dose, the high content screening(HCS) cell imaging analysis was applied to test the phagocytic ability of RAW264.7 cells engulfing GFP-E. coli in vitro, and the phagocytic rates between different kinds of samples at various concentrations were calculated. Furthermore, the biological potency was measured according to the qualitative response parallel method to better evaluate the difference of TG and NTG, by translating phagocytic rates into biological potency. RESULTS: At the range of 0.695 - 5.56 mg(crude drug) •mL-1, all 11 batches samples have no toxicity. The results of HCS displayed that every sample could promote cell phagocytic activity in varying degrees at a certain concentration. RAW264.7 cells could engulf the GFP-E. coli, and the images of HCS reflected the situation of phagocytosis clearly. In addition, the results of biopotency showed that the biopotency value of different samples was between 39.56 to 100, and the average biopotency value of TG was significantly higher than NTG (P < 0.05). CONCLUSION: In vitro experiments showed that all the samples could promote the phagocytic activity of RAW264.7 cells at different degrees. And the average biopotency value of TG was significantly higher than NTG, which is consistent with the saying that "TG has a better quality".

Activating blood biological potency assay and chemical fingerprint chromatogram applied to quality evaluation of rhubarb / 药学学报

The biological potency assay and chemical fingerprint chromatogram were applied to quality evaluation of rhubarb. Using the biological potency as indicators, we evaluated the differences in quality of multiple batches of rhubarbs and related products. Using the platelet aggregation analyzer, we determined platelet aggregation rate in the different rhubarbs preparations, and calculated the biological potency based on the simplified probit principle. UPLC was adopted to establish the fingerprint spectra for rhubarbs. The spectral efficiency correlation analysis between chromatograms and biological potencies were conducted using the double variables of SPSS 22.0 software. We used three chemical composition to verify the potency. The biological potency results suggest that Rheum palmatum has a more potent activity than Rheum tanguticum, and wine-treated rhubarb had a higher potentcy than charred. We identified 10 elements in the Fingerprint Spectrum. The relevant elements including rhein-8-O-β-D-glucoside, emodin-8-O-β-D-glucoside and rhein have the strongest activity in the inhibition of platelet aggregation. In conclusion, this study provides a analytical method for rhubarb biological potency based on determination of the maximum antagonism rate model. The rhein may be the effective substance. It may serve as a reference in the quality control of wine processed rhubarb products.

Study on quality control methods of Fuzheng Xiaoji capsule based on biological detection method / 中国生化药物杂志

Objective To establish biological detection method for Fuzheng Xiaoji capsule.MethodsTaken Radix astragali,Hedyotis diffusa Willd, Fructus Polygoni orientalis and rhubarb in Fuzheng Xiaoji capsule as targets, the antimicrobial activity of the single fried mixture of Radix astragali,Hedyotis diffusa Willd,Fructus Polygoni orientalis and rhubarb to 4 kinds of standard strains(Staphylococcus aureus, Salmonella, Escherichia coli and Bacillus subtilis) were inspected.The standard curve by susceptible strains of the inhibition zone diameter and logarithm of the concentration was established,and this test also determined the biological potency of different batches of Fuzheng Xiaoji capsule according to the dose-response curve.ResultsThe single fried mixture of Radix astragali,Hedyotis diffusa Willd, Fructus Polygoni orientalis and rhubarb have strong anti-bacterial effect to the above four standard strains.There was a good linear relationship between logarithmic dose and response effect when the concentration in the range of 0.029-0.136g/mL(r=0.9583).ConclusionBiological potency detection method can be combined with traditional analysis methods to control the quality of Fuzheng Xiaoji capsule.

Study on antiplatelet aggregation active components of Erigeron breviscapus in vitro based on spectrum-effect correlation analysis / 中草药

Objective: To explore the active components of antiplatelet aggregation of Erigeron breviscapus, chemical fingerprints and bioactivity detection were used to carry out spectrum-effect correlation analysis. Methods: A fingerprinting method was established by ultra-high performance liquid chromatography with ultraviolet detection (UPLC-UV) and then used for fingerprinting of different batches of E. breviscapus. The antiplatelet aggregation biopotency of different batches of E. breviscapus was tested, and the possible active substances were deduced based on spectrum-effect correlation analysis. Furthermore, all five compounds were verified by antiplatelet aggregation in vitro, and the contribution value of relative activity of the five compounds was calculated according to the difference of the contents of five kinds of monomer compounds in E. breviscapus. Results: Through the spectrum-effect correlation analysis of chemical fingerprints and antiplatelet aggregation biopotency of E. breviscapus, five chromatographic peaks with higher bioactivity correlation coefficient were screened and identified, including chlorogenic acid, caffeic acid, scutellarin, isochromic acid A, and ischlorogenic acid C. Further in vitro experiments showed that the five compounds had different levels of antiplatelet aggregation at same concentration (inhibition rate: 16.5%-85.5%). The sequence of the relative activity contribution is scutellarin > ischlorogenic acid C > caffeic acid > ischlorogenic acid A > chlorogenic acid. In terms of activity contribution of five compounds, chlorogenic acid C and scutellarin was larger than other compounds. Conclusion: A method for the determination of antiplatelet aggregation biopotency of E. breviscapus in vitro was established. Moreover, scutellarin and ischlorogenic acid C are the main active substances of E. breviscapus in the aspect of antiplatelet aggregation in vitro.

Estudo colaborativo nacional para o estabelecimento do material de referência de trabalho da vacina contra sarampo, caxumba e rubéola: evolução para a autossuficência na produção nacional da vacina tríplice viral / National collaborative study for establishing the working reference material of the vaccine against measles/mumps and rubella: evolution to the self-sufficiency in the national production of triple viral vaccine

A monografia farmacopeica da vacina tríplice viral (sarampo/caxumba/rubéola) exige a validação de desempenho do ensaio de potência utilizando-se apropriado material de referência (MR). Com o intuito de estabelecer o primeiro MR de trabalho (MRT) nacional para a vacina tríplice viral, foi realizado o estudo colaborativo nacional com a participação de duas únicas instituições que executam o ensaio de potência desta vacina, o Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos, produtor nacional) e o Instituto Nacional de Controle de Qualidade em Saúde.O material candidato (cMRTBio), preparado pelo produtor, foi avaliado pelos laboratórios participantes utilizando-se as respectivas metodologias in-house de determinação de potência. O cMRTBio foi considerado apropriado como MR in-house por estar em concordância com as especificações recomendadas nas normativas de compêndios, a saber: variações intra- (< 5 %), inter-ensaios (< 10 %) e entre laboratórios (< 10 %) abaixo dos limites aceitáveis; e potência estimada (log10 CCID50/DH) em 3,72 para sarampo, 4,80 para caxumba e 3,70 para rubéola. Este trabalho reflete o compromisso do único produtor nacional da vacina tríplice viral com a saúde pública, descrevendo-se a expansão da tecnologia, o cumprimento às diretrizes internacionais, o cuidado com o controle da qualidade e culminância para a autossuficiência nacional na produção de vacinas.

The pharmacopoeia monograph for the measles/mumps and rubella (MMR) triple vaccine demands to perform the validation of the potency assay by using the suitable reference material (MR). Aiming at establishing the firstwork MR (MRT) for the MMR triple vaccine, a national collaborative study was performed with the participation of the two unique national institutions working on the vaccine potency evaluation test, the Imunobiological Technology Institute (Bio-Manguinhos, national manufacturer) and the National Institute for Quality Control in Health. The candidate product (cMRTBio) prepared by the manufacturer was evaluated by the participant laboratories by employing the respective in-house methodologies for determining the potency. The cMRTBio was considered suitable as in-house MR, according to the specifications based on the normative compendia, being the intra-assay (< 5 %), inter-assay(< 10 %) and between laboratories variations (< 10 %) below the acceptable limits, and the estimate potency(log10 CCID50/DH) in 3.72 for measles, 4.80 for mumps and 3.70 for rubella. This study reflects the commitment of the unique national MMR vaccine producer to the public health, describing the expansion of technology,the compliance with international guidelines and the careful quality control, leading to the national self-sufficiency in the vaccine production.

Study on Integral Dissolution Model Based on Biological Potency for Compound Chinese Materia Medica / 中草药·英文版

Objective: To investigate the integral dissolution model based on biological potency in order to evaluate the dissolution of Compound Chinese materia medica (CCMM) in vitro. Methods: The contents of paeoniflorin, phillyrin, ginsenoside Rg1, and adenosine of ten batches of Compound Biejia Ruangan Tablet (CBRT) were determined at different times. The self-defined weighting coefficient based on the contents has been created to establish the integral dissolution model. In addition, the biological potency of CBRT was measured by MTT assay. Then, the f2 similar factor was used to evaluate the similarity of the batches. Results: Compared with batch a, some batches' f2 values of paeoniflorin and adenosine were less than 50, while f2 values of ginsenoside Rg1, phillyrin, and integral component were more than 50. Likewise, ginsenoside Rg1, phillyrin, and integral component were all in good correlation with biological dissolution. Conclusion: The results of the integral dissolution based on biological test of CBRT demonstrate that the bioassay method may be a promising supplement for its quality evaluation.

Antiasthma biological potency applied for Ephadra Herba quality evaluation / 中草药

Objective: To study the anti-asthma action and biological potency differences of Ephedra Herba from 37 different habitats. Methods: Taking the isolated tracheal smooth muscle from guinea pig to establish antiasthma pharmacological model, the ephedra decoction at different concentration was added to the sink by cumulative metrology method, antispasmodic percentage and biological potency value of Ephedra Herba from different habitats were calculated and the cluster analysis for the biological potency value of Ephedra Herba from different habitats was conducted. Results: Ephedra Herba had the antispasmodic acction to the isolated tracheal smooth muscle of guinea pig caused by histamine phosphate, and compared with the control medicine, Ephedra Herba from 20 habitats showed significant differences (P 0.05); Compared with the control medicine, the antiasthma biological potency of Ephedra Herba from 37 different habitats existed significant or very significant differences (P < 0.05, 0.01), and the antiasthma biological potency value was 22.35-489.04 U/g, while FL% was 13.15%-38.97%, which revealed that the high potency level had a difference of 21.88 times from the low one. And the antiasthma biological potency of Ephedra Herba had lowly significant correlation differences with the total determination of ephedrine and D-pseudephedrine; The Ephedra Herba with different antiasthma biological potency could be identified by cluster analysis method, in which the biological potency from 24 ones in total of 37 Ephedra Herba was higher than that in the control medicine and accounted for 64.86% of the total samples. Conclusion: The anti-asthma biological potency value of Ephedra Herba could quantitatively evaluate the quality of Ephedra Herba from different habitats.