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Preparation of the Molecular Mass Calibration Reference Standard for Molecular Weight Hepairn

Author: DuZuoZuo
Tutor: CuiHuiZuo
School: Shandong University
Course: Microbial and Biochemical Pharmacy
Keywords: Low molecular weight heparin Molecular weight Molecular weight reference High Performance molecular exclusion
CLC: R927.2
Type: Master's thesis
Year: 2011
Downloads: 70
Quote: 0
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Abstract

Low molecular weight heparin (low molecular weight heparin, LMWH) is prepared from heparin by chemical or physical degradation or heparanase grading method heparin fragment or component, having an average molecular weight of 8000 or less, less than 8000 components not less than 60 %, the anti-FXa activity is not less than 70IU/mg, Anti-FXa / anti-Flla not less than 1.5. LMWH and heparin body consistent with the structure of disaccharide units is a mixture of polysaccharide chains composed of iduronic acid or glucuronic acid and glucosamine, as well as their sulfated and acetylated derivatives (1 → 4) glycosidic linkage is made. Clinical practice has proved in the past 30 years, LMWH is an excellent anticoagulant antithrombotic drugs, compared with Heparin antithrombotic activity equivalent, but more secure. China is the world heparin largest producer, but the current situation is heparin production to more than 80% of exports in the form of bulk drugs, heparin API export after processing into a higher price LMWH preparations sold back to return. Although China has more than ten manufacturers LMWH preparations, but LMWH quality standards compared with foreign countries was significantly lower, mainly to meet the definition of low-molecular-weight heparin molecular weight and activity requirements, according to the classification process, and prepared by different processes of LMWH have different end structures, the type of process-related impurities, the type of average molecular weight and distribution, which affect the activity of the low molecular weight heparin and in vivo processes. Therefore, according to foreign standards, and combined with national conditions as soon as possible to improve the domestic LMWH quality standards is necessary. Molecular weight distribution is an important indicator of LMWH quality standards. Currently there are no domestic LMWH molecular weight determination with reference only European Pharmacopoeia (European Pharmacopoeia, EP) LMWH molecular weight determination by reference, WHO LMWH molecular weight determination with reference (International Reference Preparation LMWH for molecular weight calibration) and the United States Pharmacopeia (United States Pharmacopoeia, USP) enoxaparin sodium molecular weight determination with reference substance. This topic is committed to the development of domestic LMWH molecular weight of the reference developed a series of narrow molecular weight distribution of LMWH molecular weight standard, and with a wide distribution of standard sheet width the LMWH molecular weight distribution of reference, and two reference control with the WHO and USP The article of the measurement result of the comparison. The results and conclusions of this study are the following aspects: one subject developed a series of narrow molecular weight distribution of LMWH molecular weight determination with the reference series the the homemade narrow distribution of of LMWH molecular weight reference substance Mp: 1400,1850,2328,2863,5714 7768,10443. 7 homemade narrow distribution of molecular weight standard, and USP Enoxaparin Sodium Molecular Weight reference calibration curve is essentially coincident with the determination of two different batches, respectively, with the homemade narrow distribution of the reference and the USP Enoxaparin Sodium Molecular Weight reference enoxaparin sodium, USP Enoxaparin Sodium Molecular Weight The molecular weight of the System Suitability Reference Sample and EP of LMWH molecular weight Reference Sample times, and results showed that the two Reference Sample Mw of each of the samples was measured away from not more than 50, Reference Preparation of a series of narrow molecular weight distribution samples as LMWH molecular weight determination with reference substance. The molecular weight of less than 6000 five narrow distribution of molecular weight of LMWH preparation method of determination of reference (1400,1850,2328,2863,5714) enoxaparin sodium as raw material, by the separation of Biogel P10. On Biogel P10 separation enoxaparin sodium showing that small molecular weight fraction jagged peaks were collected corresponding peak can be obtained by a narrow distribution of a series of reference 6000. 6000 two narrow distribution of molecular weight of LMWH measurement reference (7768,10443) based the homemade (chemical beta elimination) MMWH (Mp8929) as raw material, obtained by Biogel P10 secondary separation. The subject tried to SuperdexG75, SephadexG75, UltragelACA44 chromatography media separation heparin sodium Sephadex G75 chromatography media separation of 55% graded alcohol precipitation heparin samples and ultrafiltration separation between 10kDa and 5kDa, component, and found regardless of the choice of raw materials and chromatography media can not get a similar separation the enoxaparin sodium jagged peaks and the samples yield obtained using the above method and RI values ??are lower response. The final choice of the yield and RI the corresponding value higher MMWH as raw material, the Biogel P10 secondary separation component as 7000 to 11100 narrow distribution of LMWH molecular weight reference. Prepared reference USP Enoxaparin Sodium narrow distribution of molecular weight reference comparing peak is still wide, SAX-HPLC, IPRP-HPLC further separation homemade LMWH molecular weight of 7000 to 11100 narrow distribution reference, hope can get the ingredients single component. Result, SAX-HPLC and IPRP-HPLC can be the Enoxaparin Sodium small molecular weight components further separated into a single component, but can not be separated further component on the molecular weight of 7000 to 11100. 2 developed a wide distribution of LMWH molecular weight with a wide distribution of standard table reference. Degradation of heparin sodium heparin sodium enzyme I get a wide distribution of LMWH as broad LMWH molecular weight reference, with a wide distribution of the standard table by the molecular weight of from 600 to 18,000 18 molecular weight corresponding cumulative percentage area, based on the wide distribution Sample Table 18 points in the cumulative percentage of area to determine corresponding to the wide distribution of the retention time, the retention time table corresponding to the molecular weight of the fitting carried out three times the logarithm of the curve equation to obtain the calibration curve and the calculated molecular weight. Homemade wide molecular weight distribution of the Reference Sample WHO molecular weight of the reference UV and RI Spectrum Similar almost coincident with a calibration curve obtained by the two reference substance. 7 LMWH heparin molecular weight were measured with the WHO the LMWH molecular weight standard, and homemade wide distribution of molecular weight reference both the measurement result is not 50, the prepared samples can be used as a broad the LMWH molecular weight reference determination LMWH molecular weight. EP - LMWH molecular weight reference standards RI and UV associated with a dedicated GPC software, just the subject prepared by the two LMWH molecular weight reference RI detector and GPC software, the calculation method is easier. This project successfully developed a molecular weight of from 1000 to 11100 series of narrow molecular weight distribution LMWH molecular weight standard, and with a wide distribution of the standard table wide distribution of molecular weight reference, for LMWH molecular weight determination of the localization of the reference laid the foundation.

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CLC: > Medicine, health > Pharmacy > Pharmacopoeia Codex ( formulary ),drug identification > Drug identification > Determination of drug content
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