低分子量肝素核磁共振波谱法的定性鉴别方法研究
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摘要
目的:建立低分子量肝素核磁共振波谱法(NMR)的定性鉴别方法。方法:一维~(13)C-NMR实验是在装配5 mm BBO探头的Bruker Ascend-500核磁共振谱仪上采集。采样时间1.1秒;弛豫延长时间(D1)1秒;测量温度40℃;谱宽δ236。结果:通过不同类型低分子量肝素标准品和市售产品的核磁共振一维碳谱精细结构信息比较发现,不同类型的低分子量肝素都可以通过碳谱特征信号进行区分。结论:核磁共振一维碳谱定性鉴别低分子量肝素类型的方法,专属性强,简单方便,是鉴别低分子量肝素较好的方法。
Objective:To establish an NMR method for the identification of low molecular weight heparin.Method:Carbon nuclear magnetic resonance(~(13)C-NMR)spectra were collected by using a Bruker Ascend 500 spectrometer with 5 mm BBO probe.The measurement parameters setting were:acquisition time 1.1 s,relaxation delay(D1)1 s,temperature 40 ℃,and spectrum width δ 236.Result:By comparing the ~(13)C-NMR spectra of low molecular weight reference standard and purchased heparin,there were some unique signals in the ~(13)C-NMR spectra,which could be used to identify different types of low molecular weight heparin.Conclusion:1D ~(13)C-NMR spectroscopy is a specific,simple and convenient method for the identification of low molecular weight heparin.
Objective:To establish an NMR method for the identification of low molecular weight heparin.Method:Carbon nuclear magnetic resonance(~(13)C-NMR)spectra were collected by using a Bruker Ascend 500 spectrometer with 5 mm BBO probe.The measurement parameters setting were:acquisition time 1.1 s,relaxation delay(D1)1 s,temperature 40 ℃,and spectrum width δ 236.Result:By comparing the ~(13)C-NMR spectra of low molecular weight reference standard and purchased heparin,there were some unique signals in the ~(13)C-NMR spectra,which could be used to identify different types of low molecular weight heparin.Conclusion:1D ~(13)C-NMR spectroscopy is a specific,simple and convenient method for the identification of low molecular weight heparin.
引文
[1]CASU B,NAGGI A,TORRI G.Re-visiting the structure of heparin[J].Carbohydr Res,2015:403(1):60
[2]CASU B.Heparin structure[J].Haemostasis,1990,20(Suppl 1):62
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[6]HULL RD,LIANG J,BERGQVIST D,et al.Benefit-to-harm ratio of thromboprophylaxis for patients undergoing major orthopaedic surgery.A systematic review[J].Thromb Haemost,2014,111(2):199
[7]HIRSH J,WARKENTIN TE,SHAUGHNESSY SG,et al.Heparin and low-molecular-weight heparin:mechanisms of action,pharmacokinetics,dosing,monitoring,efficacy,and safety[J].Chest,2001,119(1 Suppl):64S
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[10]董学畅,张淑桂,杨素仙.低分子肝素及其制备[J].云南化工,2000,27(1):44DONG XC,ZHANG SG,YANG SX.The low molecular weight heparin and its preparation[J].Yunnan Chem Technol,2000,27(1):44
[11]SHIVELY JE,CONRAD HE.Formation of anhydrosugars in the chemical depolymerization of heparin[J].Biochemistry,1976,15(18):3932
[12]LINHARDT RJ,GUNAY NS.Production and chemical processing of low molecular weight heparins[J].Semin Thromb Hemost,1999,25(Suppl 3):5
[13]NICOLAIDES AN,FAREED J,KAKKAR AK,et al.Prevention and treatment of venous thromboembolism—International Consensus Statement[J].Inter Angiol:J Inter Union Angiol,2013,32(2):111
[14]GUERRINI M,BECCATI D,SHRIVER Z,et al.Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events[J].Nat Biotechnol,2008,26(6):669
[15]MARUYAMA T,TOIDA T,IMANARI T,et al.Conformational changes and anticoagulant activity of chondroitin sulfate following its O-sulfonation[J].Carbohydr Res,1998,306(1-2):35
[16]USP 36[S].2017:3979
[17]EP 8.0[S].2014:0828
[18]彭向宇.百特事件后我国肝素企业首位度分析[J].肉类工业,2014,394(2):108PENG XY.The analysis of Baxter incident after the first heparin enterprises in China[J].Meat Ind,2014,394(2):108
[19]张琪,王爱法,朱红波,等.低分子量肝素核磁共振鉴别方法的研究[J].中国药品标准,2015,16(2):123ZHANG Q,WANG AF,ZHU HB,et al.Study of NMR identification method for low molecular weight heparin[J].Drug Stand China,2015,16(2):123
[20]SANDERSON PN,HUCKERBY TN,NIEDUSZYNSKI IA.Chondroitinase ABC digestion of dermatan sulphate.N.m.r.spectroscopic characterization of the oligo-and poly-saccharides[J].Biochem J,1989,257(2):347
[21]MULLOY B,FORSTER MJ,JONES C,et al.NMR and molecularmodelling studies of the solution conformation of heparin[J].Biochem J,1993,293(Pt 3):849
[22]MULLOY B,MOURAO PA,GRAY E.Structure/function studies of anticoagulant sulphated polysaccharides using NMR[J].J Biotechnol,2000,77(1):123
[2]CASU B.Heparin structure[J].Haemostasis,1990,20(Suppl 1):62
[3]LEE GM,AREPALLY GM.Heparin-induced thrombocytopenia[J].Hematology Am Soc Hematol Educ Program,2013,2013:668
[4]REES S.How common medicines can affect bone health[J].Nurs Times,2014,110(32-33):25
[5]LIU Z,JI S,SHENG J,et al.Pharmacological effects and clinical applications of ultra low molecular weight heparins[J].Drug Discov Therap,2014,8(1):1
[6]HULL RD,LIANG J,BERGQVIST D,et al.Benefit-to-harm ratio of thromboprophylaxis for patients undergoing major orthopaedic surgery.A systematic review[J].Thromb Haemost,2014,111(2):199
[7]HIRSH J,WARKENTIN TE,SHAUGHNESSY SG,et al.Heparin and low-molecular-weight heparin:mechanisms of action,pharmacokinetics,dosing,monitoring,efficacy,and safety[J].Chest,2001,119(1 Suppl):64S
[8]KLEINSCHMIDT K,CHARLES R.Pharmacology of low molecular weight heparins[J].Emerg Med Clin North Amer,2001,19(4):1025
[9]QUARTERMAIN D,LI Y,JONAS S.Enoxaparin,a low molecular weight heparin decreases infarct size and improves sensorimotor function in a rat model of focal cerebral ischemia[J].Neurosci Lett,2000,288(2):155
[10]董学畅,张淑桂,杨素仙.低分子肝素及其制备[J].云南化工,2000,27(1):44DONG XC,ZHANG SG,YANG SX.The low molecular weight heparin and its preparation[J].Yunnan Chem Technol,2000,27(1):44
[11]SHIVELY JE,CONRAD HE.Formation of anhydrosugars in the chemical depolymerization of heparin[J].Biochemistry,1976,15(18):3932
[12]LINHARDT RJ,GUNAY NS.Production and chemical processing of low molecular weight heparins[J].Semin Thromb Hemost,1999,25(Suppl 3):5
[13]NICOLAIDES AN,FAREED J,KAKKAR AK,et al.Prevention and treatment of venous thromboembolism—International Consensus Statement[J].Inter Angiol:J Inter Union Angiol,2013,32(2):111
[14]GUERRINI M,BECCATI D,SHRIVER Z,et al.Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events[J].Nat Biotechnol,2008,26(6):669
[15]MARUYAMA T,TOIDA T,IMANARI T,et al.Conformational changes and anticoagulant activity of chondroitin sulfate following its O-sulfonation[J].Carbohydr Res,1998,306(1-2):35
[16]USP 36[S].2017:3979
[17]EP 8.0[S].2014:0828
[18]彭向宇.百特事件后我国肝素企业首位度分析[J].肉类工业,2014,394(2):108PENG XY.The analysis of Baxter incident after the first heparin enterprises in China[J].Meat Ind,2014,394(2):108
[19]张琪,王爱法,朱红波,等.低分子量肝素核磁共振鉴别方法的研究[J].中国药品标准,2015,16(2):123ZHANG Q,WANG AF,ZHU HB,et al.Study of NMR identification method for low molecular weight heparin[J].Drug Stand China,2015,16(2):123
[20]SANDERSON PN,HUCKERBY TN,NIEDUSZYNSKI IA.Chondroitinase ABC digestion of dermatan sulphate.N.m.r.spectroscopic characterization of the oligo-and poly-saccharides[J].Biochem J,1989,257(2):347
[21]MULLOY B,FORSTER MJ,JONES C,et al.NMR and molecularmodelling studies of the solution conformation of heparin[J].Biochem J,1993,293(Pt 3):849
[22]MULLOY B,MOURAO PA,GRAY E.Structure/function studies of anticoagulant sulphated polysaccharides using NMR[J].J Biotechnol,2000,77(1):123