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A facile colorimetric protamine titration method.

C L Teng1, V C Yang

  • 1College of Pharmacy, University of Michigan, Ann Arbor 48109-1065.

The Journal of Laboratory and Clinical Medicine
|April 1, 1989
PubMed
Summary
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A new colorimetric method using azure A dye offers a rapid and accurate alternative to traditional clotting assays for protamine titration. This faster assay helps determine precise protamine doses for heparin neutralization and aids in preparing samples for coagulation tests.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Pharmacology

Background:

  • Heparin is an anticoagulant requiring neutralization by protamine.
  • Traditional protamine titration methods rely on time-consuming clotting assays.
  • Accurate protamine dosing is crucial for managing heparin therapy and its reversal.

Purpose of the Study:

  • To develop a facile, colorimetric method for protamine titration.
  • To replace the traditional clotting assay with a rapid colorimetric assay.
  • To provide a reliable tool for estimating protamine doses for heparin neutralization.

Main Methods:

  • Utilized the competitive binding of protamine and azure A dye to heparin.
  • Employed azure A dye as a colorimetric indicator for titration.

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  • Developed a diagnostic kit for rapid determination of titration end points.

    • Main Results:

    • The colorimetric method provides accuracy comparable to traditional clotting assays.
    • The assay significantly shortens processing time, with results available in under 5 minutes.
    • The method is effective for both protamine titration and heparin titration (reverse assay).

    Conclusions:

    • The new colorimetric protamine titration method is a rapid, accurate, and reliable tool for clinical use.
    • It simplifies heparin neutralization dose estimation and facilitates sample preparation for coagulation tests.
    • The assay also offers a method for assessing heparin reversal and managing protamine overdose.