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Current Pharmaceutical Analysis

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ISSN (Print): 1573-4129
ISSN (Online): 1875-676X

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Research Article

Determination of Thrombin Activity by Reversed-phase High Performance Liquid Chromatography

Author(s): Liang Zhang, Dehui Yang and Ye Liu*

Volume 19, Issue 3, 2023

Published on: 30 December, 2022

Page: [216 - 221] Pages: 6

DOI: 10.2174/1573412919666221124110031

Price: $65

Abstract

Background: A reversed-phase high-performance liquid chromatographic method for determining thrombin activity is described. The improved method can provide a high resolution between substrate Nα-P-toluenesulfonyl-L-arginine methyl ester (TAME) and the product of enzyme reaction, Nα-P-toluenesulfonyl-L-Arginine (TA).

Methods: When the substrate concentration was 5 mg/mL, the Michaelis equation approached the zero-order reaction, and the thrombin enzyme activity must have a good linear relationship with the consumption of TAME or the yield of TA.

Results: The method is suitable for quantitatively analyzing thrombin activity in formulation or serum.

Conclusion: Three batches of thrombin lyophilized preparations were determined for activity.

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