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Determination of low-molecular-weight heparins and their binding to protamine and a protamine analog using polyion-sensitive membrane electrodes.
Anal Biochem 1999; 266(1):116-24AB

Abstract

A polycation-sensitive membrane electrode based on the ion-exchanger dinonylnaphthalene sulfonate has previously been developed and used as an end-point detector for the determination of unfractionated heparin in whole blood samples via simple potentiometric titration with protamine. Herein, we report the application of the same methodology for the quantitation of a commercial low-molecular-weight heparin (LMWH) preparation (Fragmin) in whole blood samples at concentrations up to 2 U/ml. Further, an analogous polyanion (heparin)-sensitive electrode is used to estimate the binding constants between protamine and various LMWH preparations. The equilibrium constants (Keq) and the number of binding sites per mole of heparin (n) are determined by recasting the data in the form of a Scatchard plot. Results show that the average molecular weight and molecular weight distribution of the LMWH preparation are important parameters affecting their binding with protamine. Comparable binding constants are obtained for the same LMWH preparations titrated with a synthetic protamine analog, [+18RGD] [acetyl-EA(R2A2R2A)4R2GRGDSPA-NH2].

Authors+Show Affiliations

Department of Chemistry, University of Michigan, Ann Arbor, Michigan, 48109, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

9887220

Citation

Ramamurthy, N, et al. "Determination of Low-molecular-weight Heparins and Their Binding to Protamine and a Protamine Analog Using Polyion-sensitive Membrane Electrodes." Analytical Biochemistry, vol. 266, no. 1, 1999, pp. 116-24.
Ramamurthy N, Baliga N, Wakefield TW, et al. Determination of low-molecular-weight heparins and their binding to protamine and a protamine analog using polyion-sensitive membrane electrodes. Anal Biochem. 1999;266(1):116-24.
Ramamurthy, N., Baliga, N., Wakefield, T. W., Andrews, P. C., Yang, V. C., & Meyerhoff, M. E. (1999). Determination of low-molecular-weight heparins and their binding to protamine and a protamine analog using polyion-sensitive membrane electrodes. Analytical Biochemistry, 266(1), pp. 116-24.
Ramamurthy N, et al. Determination of Low-molecular-weight Heparins and Their Binding to Protamine and a Protamine Analog Using Polyion-sensitive Membrane Electrodes. Anal Biochem. 1999 Jan 1;266(1):116-24. PubMed PMID: 9887220.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of low-molecular-weight heparins and their binding to protamine and a protamine analog using polyion-sensitive membrane electrodes. AU - Ramamurthy,N, AU - Baliga,N, AU - Wakefield,T W, AU - Andrews,P C, AU - Yang,V C, AU - Meyerhoff,M E, PY - 1999/1/15/pubmed PY - 1999/1/15/medline PY - 1999/1/15/entrez SP - 116 EP - 24 JF - Analytical biochemistry JO - Anal. Biochem. VL - 266 IS - 1 N2 - A polycation-sensitive membrane electrode based on the ion-exchanger dinonylnaphthalene sulfonate has previously been developed and used as an end-point detector for the determination of unfractionated heparin in whole blood samples via simple potentiometric titration with protamine. Herein, we report the application of the same methodology for the quantitation of a commercial low-molecular-weight heparin (LMWH) preparation (Fragmin) in whole blood samples at concentrations up to 2 U/ml. Further, an analogous polyanion (heparin)-sensitive electrode is used to estimate the binding constants between protamine and various LMWH preparations. The equilibrium constants (Keq) and the number of binding sites per mole of heparin (n) are determined by recasting the data in the form of a Scatchard plot. Results show that the average molecular weight and molecular weight distribution of the LMWH preparation are important parameters affecting their binding with protamine. Comparable binding constants are obtained for the same LMWH preparations titrated with a synthetic protamine analog, [+18RGD] [acetyl-EA(R2A2R2A)4R2GRGDSPA-NH2]. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/9887220/Determination_of_low_molecular_weight_heparins_and_their_binding_to_protamine_and_a_protamine_analog_using_polyion_sensitive_membrane_electrodes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(98)92947-5 DB - PRIME DP - Unbound Medicine ER -