National Collaborative Study to Establish the 1st National Standards of Heparin Molecular Weight Calibrant and Heparin for System Suitability of Molecular Weight Determinations / 中国药学杂志

OBJECTIVE: Molecular weight determinations of heparin sodium and heparin calcium were added in Chinese Pharmacopoeia (2015).To establish the 1st national standards of heparin molecular weight calibrant and heparin for system suitability of molecular weight determinations were needed to establish. METHODS: An national collaborative study involving twelve laboratories had taken place, organized by National Institutes for Food and Drug Control(NIFDC)to provide supporting data for the establishment of the 1st batches of heparin molecular weight calibrant and heparin for system suitability of molecular weight determinations standard. The method of heparin molecular weight determinations in Chinese Pharmacopoeia (2015) was used in the national collaborative study. The USP heparin sodium molecular weight calibrant RS (F0L483) was used as molecular weight calibrant. The USP heparin sodium identification RS(G1L413) was used as the standard of system suitability. The candidate national standards of heparin molecular weight calibrant (140819-201501) and the candidate heparin for system suitability of molecular weight determination standard(140818-201501) were tested in the study. RESULTS: To be calculated the cumulative percent of peak area at the 20 molecular points from 5 000-42 000 of the candidate national standard of heparin molecular weight calibrant (140819-201501). Based on the statistical analysis, the candidate gave low intra-and inter-laboratories variations. In laboratories, standard deviations (SD) of two laboratories ranged from 1% to 2%,others were less than 1%.Between laboratories, SD were all less than 1%,relative standard deviation(RSD) was all less than 10%. The intra-lab SD of the test to determine the molecular weight of the candidate heparin for system suitability of molecular weight determinations standard(140818-201501) was less than 180,except Lab 2. The inter-lab SD was 180. The RSD was 1.1%. CONCLUSION: After the examination of the expert committee on pharmaceutical standardization, the candidate (140819-201501) is approved as the first national standard of heparin molecular weight calibrant, provides the broad standard table. The candidate(140818-201501) is approved as the first national standard of heparin for system suitability of molecular weight determinations, with an assigned molecular weight (Mw) of 16 200. The two national standards can be used in the test of heparin molecular weight determinations in Chinese Pharmacopeia(2015).

Optimization of extraction process of Codonopsis pilosula polysaccharides and study on its composition / 中草药

Objective: To optimize the extraction process of polysaccharide from Codonopsis pilosula and determine the monosaccharide composition and molecular weight distribution, in order to provide the basis for further separation of C. pilosula polysaccharide. Methods: The content of polysaccharide in C. pilosula was determined by phenol sulfuric acid method, the extraction process of polysaccharide was optimized by orthogonal test. C. pilosula polysaccharides were prepared from crude polysaccharides by deproteinization, decoloration, dialysis, and lyophilization, then monosaccharide composition and mean molecular mass of C. pilosula polysaccharides were analyzed by high performance liquid chromatography (HPLC) and high performance gel permeation chromatography (HPGPC). Results: The extraction temperature was 85 ℃, the extraction time was 1.5 h per time, twice, and solid to liquid ratio was 1:12. Under these conditions, the yield of polysaccharides was 22.57%. The polysaccharides were consisted by glucuronic acid, aminogalactose, xylose, and small quantities of mannose, the average molecular mass was 21 498. Conclusion: This study provides a theoretical basis for the classification and activity of polysaccharide from C. pilosula.

Determination of Weight Average Molecular Weight and Molecular Weight Distribution of Ferric Carboxy-maltose by HPGPC / 中国药房

OBJECTIVE:To establish a method for the determination of weight average molecular weight(Mw)and molecular weight distribution (D) of ferric carboxymaltose. METHODS:HPGPC method was adopted to detect the Mw and D of 3 batches of Ferric carboxymaltose injection (imported) and its raw material (self-made). The determination was performed on TSK G4000 PWXL column with 0.1% sodium azide solution with the flow rate of 0.5 ml/min. The detector was refractive index detector;the column temperature was set at 35 ℃,and sample size was 20 μl. The results were calculated with GPC software. RESULTS:RSDs of precision,stability and reproducibility tests were all lower than 3.0%(n=6);Mw and D of 3 imported samples were 157 667 and 1.30;those of self-made samples were 162 000 and 1.42. CONCLUSIONS:The method has high precision,good stability,repeat-ability and durability. It can be used for the determination of Mw and D of ferric carboxymaltose.

Determination of molecular weight and weight distribution in Iron Maltose Syrup by HPGPC / 中国生化药物杂志

Objective To establish a method for determining the molecular weight (Mw) and molecular weight distribution of Iron Maltose Syrup. Methods HPGPC was used; PSS HEMA was used as column.Detector was differential refraction detector.Mobile phase was phosphate buffer solution (pH6.8) at 0.5 mL/min, column temperature was 45℃.Results The Mw of 3 batches of Iron Maltose Syrup were 45000-47000 Da with good linearity, precision and reproducibility.Conclusion The method is simple, reliable and accepted by the specification for controlling the molecular weight and weight distribution of Iron Maltose Syrup.

Research on the determination of molecular weight distribution and the peptide content of Ganlong capsule / 中国生化药物杂志

Objective To establish the molecular weight distribution of Ganlong capsule by HPSEC the content of the peptide determined by Lowry and Methods The superdex peptide 10/300 GL (10 mm ×300 mm) column was used.The pH=6.0 and phosphate buffer of 0.05 mol/L was used as the mobile phase, containing 0.1 mol/L NaCl.The flow rate was set at 0.7 mL/min;The column temperature was 25℃;The detection wavelength was 214 nm.Results The content of the peptide ranged from 0.08 mg to 0.4 mg ( r =0.9996 ) .The RSDs of measurement precision of molecular weight and content were 0.08% and 0%(n=6), respectively.The RSDs of the repeatability were 1.3% and 1.1%(n=6);The regression equation of standard material was logMr =5.1455 -0.0871tR, r =0.9983,the relative molecular weight ranged from 2.68 ×102 Da ~5.73 ×103 Da(r =0.9983). Conclusion on The method is simple and rapid for determining the peptide content and the molecular weight distribution of Ganlong capsule.It can be used quality control method for Ganlong capsule.

Research on the determination of molecular weight distribution of anti-HBV placenta transfer factor injection / 中国生化药物杂志

Objective To establish the molecular weight distribution of anti-HBV placenta transfer factor injection (PSTF) by electrophoresis, HPLC and MS.Methods Using the methods of SDS-PAGE, HPSEC, MALDI-TOF-MS to test the molecular of PSTF.Results The Molecular was 8000 Da by SDS-PAGE.There were 5026.67,6783.44,7496.42,8736.55 Da components in PSTF by HPSEC.The main component molecular was 2972 Da and the maximum molecular component was 8194 Da.Conclusion HPSEC is simple and rapid to determine the maximum component molecular of PSTF.

Distribuição de massa molar em um biorreator com membrana para tratamento de efluente de laticínios / Molecular weight distribution in a membrane bioreactor for dairy effluent treatment

Os biorreatores com membrana (BRM) apresentam-se como um dos processos mais promissores para tratamento de águas residuárias com elevada carga orgânica, como os efluentes de laticínios, propiciando a geração de um efluente com elevada qualidade e adequado ao reuso direto ou após tratamento terciário. O objetivo desse trabalho foi avaliar o uso de BRM para tratamento de efluente de indústria de laticínios e utilizar a distribuição de massa molar da alimentação, do permeado e da fração solúvel do lodo como ferramenta para a investigação dos mecanismos de remoção dos poluentes no sistema. O BRM se mostrou um sistema viável para o tratamento do efluente em questão, apresentando eficiências de remoção de demanda química de oxigênio (DQO) e cor aparente de 98 e 99%, respectivamente. Através da distribuição de massa molar foi possível observar a alta capacidade de biodegradação e a estabilidade proporcionada pelo BRM, já que, mesmo em situações de alterações constantes nas características da alimentação, o líquido reacional sempre apresentou baixas concentrações de poluentes. Ressalta-se também a importância da membrana no sistema, uma vez que, além de permitir a retenção completa de biomassa e a operação com idades de lodo e concentração de sólidos suspensos maiores, pode proporcionar ainda a retenção de compostos que não foram biodegradados, contribuindo para a geração de um efluente tratado com alta qualidade.

The membrane bioreactor (MBR) is one of the most promising processes for the treatment of high organic load wastewaters, as dairy effluent, providing the generation of an effluent with high quality, which could be reuse directly or after tertiary treatment. The aim of this study was to evaluate a MBR to treat effluent from the dairy industry and to use molecular weight distribution of the feed, permeate and the soluble fraction of the sludge as a tool for investigating the mechanisms of pollutants removal in the system. The MBR has proven to be a viable system for the treatment of the effluent in question, with removal efficiencies of chemical oxidation demand (COD) and color of 98 and 99% respectively. Through the molar weight distribution it was possible to observe the high biodegradation capacity and stability provided by the MBR, as even in situations of constant change in feed characteristics, the mixed liquid always showed low concentrations of pollutants. It is also highlighted the importance of the membrane in the system, which, besides allowing the complete retention of biomass and operation with high solids retention time and suspended solids concentration, it can provide the retention of compounds which were not biodegraded, contributing to the generation a treated effluent with high quality.

Establishment of European calibration CRS batches 2 and 3 for low molecular weight heparin by an international collaborative study / 中国药学杂志

OBJECTIVE: An international collaborative study involving fourteen laboratories has taken place, organized by the European Directorate on Quality of Medicines (EDQM) and National Institute for Biological Standards and Control(NIBSC) to provide supporting data for the establishment of replacement batches of calibration chemical reference substance (CRS) for low molecular weight heparin (LMWH). METHODS: The study was organized in two phases: A prequalification (phase 1, performed in three laboratories in 2005) followed by an international collaboration study (phase 2). Our institute (National Institutes for Food and Drug Control, NIFDC) took part in the phase 2 study started in March 2006. The molecular mass parameters were determined for seven different LMWH samples using the current CRS (CRS1) and two batches of candidate replacement material (cCRS2 and cCRS3) with a defined number average molecular mass (Mn) of 3700 determined in phase 1. RESULTS: The calculated values of cCRS2 and cCRS3 were systematically different from the values calculated using CRS1 with its assigned Mn of 3700. Using the raw data supplied by other participants, the molecular mass parameters were recalculated using cCRS2 and cCRS3 with values for Mn of 3800 and 3900. The calculated values using these Mn values agreed more closely with those calculated using CRS1, supporting the fact that the candidates, though similar in view of the production processes, differed slightly from CRS1 in terms of molecular mass distribution. CONCLUSION: The establishment of cCRS2 and cCRS3 could be recommended with an assigned Mn value of 3 800 that is consistent with both the phase 1 results and the determination result of current CRS1.

Influence of Different Extraction and Purification Methods on Astragalus Polysaccharides and Pharmacological Evaluation / 中草药·英文版

Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides (APS) as complementary therapeutic agents prepared by different extraction and purification techniques. Methods Components of APS prepared by different extraction and purification techniques were analyzed, and these APS were used for synergy and attenuation of chemotherapy, radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice, and also used for the regulation of immune function to immunosuppression mice. Results Experimental data were analyzed by means of statistical method to get pharmaco-result: A3 (extracted by microwave assistance and purified by membrane separation) > A4 (extracted by refluxing and purified by membrane separation) > A1 (extracted by refluxing and no purification)≈ A2 (extracted by microwave assistance and no purification). There were no significant differences on pharmacodynamic action between A1 and A2. However, compared with A1 and A2,it was worth noting that A3 and A4 exhibited good pharmacodynamic action. Then A3-in and A4-in, the samples in dialyzer after dialysis, were separated and purified to get homogeneous APS, which were the principal constituents of APS in dialyzer, with the molecular weight (Mw) of 7669 and 14 142 determined by HPGPC, respectively. The average Mw of APS outside of the dialyzer, A3-out was 3102 and A4-out 3256, which were the main compositions of A3 and A4, accounted for 79.63% and 53.92%, respectively. Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity. Refluxing, microwave assistance extractions, and membrane enrichment techniques bring different cases on Mw distribution, components and pharmacodynamic action, and obviously exhibit relationship among component, Mw distribution, and pharmacological action.

DISTRIBUTION OF MOLECULAR WEIGHT OF PEPTIDE PREPARED BY COMPLEX ENZYME AND THE TUMOR INHIBITION TEST / 营养学报

Objective To observe the molecular weight distribution of soybean peptides prepared by complex enzyme hydrolysation and tumor inhibitory effect in vivo or in vitro. Method The soybean protein was enzymolyzed to soybean peptides by Flavourzyme, Alealase and neutral protease, at(50-55)℃, followed by separation, absorption,affination, ultra centrifuge and spray drying. The soybean peptides were passed Sephadex G25 column at 0.5ml/min with 0.1mol/L Tris-HCl buffer. The fragment of fluid collected was determined by UV at 280nm and HPLC. Then the molecular weight distribution was calculated. The inhibitory effect on ansplant glioma (G422) at dose 1.25, 2.5 and 5g/kg bw (ig daily for 11 d) was tested in mice and in Hep-2 in vitro. Results The peptides contents in three batches were 54.6%, 51.9%, 51.8% respeclively. The molecular weight