Physical characterization of heparin by light scattering. 1995

A K Gaigalas, and J B Hubbard, and R LeSage, and D H Atha
Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.

Dynamic and electrophoretic light scattering were used to measure the size and charge heterogeneity of a commercial preparation of heparin. For this preparation of porcine mucosal heparin (M(r) = 10-20 kDa), the diffusion coefficient was 1.2 +/- 0.5 x 10(-8) cm2/S and the mobility was 4.4 +/- 0.9 x 10(-4) cm2/Vs for an unfiltered solution at 22 degrees C in distilled water. This diffusion constant is 2 orders of magnitude smaller than expected for a molecule the size of heparin. A fast diffusion component of 5.8 +/- 1.0 x 10(-7) cm2/S, corresponding to the individual molecule, was observed in the presence of 2 M NaCl, where single molecule motion is better observed. This indicates that a portion of the heparin population is in an aggregated state, which produces a higher scattering intensity than individual heparin molecules. The weight percentage of the aggregates was 5-10 as measured by high performance exclusion chromatography. These aggregates were stable up to a temperature of 75 degrees C as measured by light scattering. This suggests that the aggregates are made up of tightly bound heparin molecules. From the diffusion coefficients we estimate the average aggregate to be made up of about 50 heparin monomers, and from the mobility, we estimate the electrophoretic charge on the aggregate to be about 1000 or about 20 electrophoretic charges for each heparin monomer. The electrophoretic light-scattering measurements also indicate that the aggregate scattering species have very similar surface charge densities resulting from the aggregates being formed from heparin molecules.(ABSTRACT TRUNCATED AT 250 WORDS)

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008027 Light That portion of the electromagnetic spectrum in the visible, ultraviolet, and infrared range. Light, Visible,Photoradiation,Radiation, Visible,Visible Radiation,Photoradiations,Radiations, Visible,Visible Light,Visible Radiations
D008433 Mathematics The deductive study of shape, quantity, and dependence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Mathematic
D002626 Chemistry, Pharmaceutical Chemistry dealing with the composition and preparation of agents having PHARMACOLOGIC ACTIONS or diagnostic use. Medicinal Chemistry,Chemistry, Pharmaceutic,Pharmaceutic Chemistry,Pharmaceutical Chemistry,Chemistry, Medicinal
D004058 Diffusion The tendency of a gas or solute to pass from a point of higher pressure or concentration to a point of lower pressure or concentration and to distribute itself throughout the available space. Diffusion, especially FACILITATED DIFFUSION, is a major mechanism of BIOLOGICAL TRANSPORT. Diffusions
D004586 Electrophoresis An electrochemical process in which macromolecules or colloidal particles with a net electric charge migrate in a solution under the influence of an electric current. Electrophoreses
D006493 Heparin A highly acidic mucopolysaccharide formed of equal parts of sulfated D-glucosamine and D-glucuronic acid with sulfaminic bridges. The molecular weight ranges from six to twenty thousand. Heparin occurs in and is obtained from liver, lung, mast cells, etc., of vertebrates. Its function is unknown, but it is used to prevent blood clotting in vivo and vitro, in the form of many different salts. Heparinic Acid,alpha-Heparin,Heparin Sodium,Liquaemin,Sodium Heparin,Unfractionated Heparin,Heparin, Sodium,Heparin, Unfractionated,alpha Heparin
D012542 Scattering, Radiation The diversion of RADIATION (thermal, electromagnetic, or nuclear) from its original path as a result of interactions or collisions with atoms, molecules, or larger particles in the atmosphere or other media. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Radiation Scattering,Radiation Scatterings,Scatterings, Radiation
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor

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