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Updated: Jul 8, 2026

Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay
Published on: September 9, 2012
The recalcified coagulation activity.
1Department of Laboratory Medicine, University Hospital, Marburg, Germany. thstief@med.uni-marburg.de
This study quantifies thrombin activity in recalcified plasma, finding that pooling plasma and specific thawing methods enhance thrombin generation. This method aids in diagnosing prothrombotic states.
Area of Science:
- Biochemistry
- Hematology
- Clinical Chemistry
Background:
- Thrombin generation following plasma recalcification is crucial for hemostasis.
- Assessing thrombin activity provides insights into coagulation dynamics and potential thrombotic states.
Purpose of the Study:
- To develop and validate a method for quantifying thrombin activity in recalcified plasma.
- To investigate factors influencing thrombin generation, including plasma pooling, anticoagulant therapy, and freeze-thaw cycles.
Main Methods:
- Citrated plasma was recalcified with calcium chloride.
- Thrombin activity was measured using chromogenic substrates after incubation at 37°C.
- Plasma samples included normal pooled plasma, patient samples (heparin/coumarin), and samples subjected to different storage conditions.
Main Results:
- Normal recalcified plasma generated 0.1-0.2 IU/ml thrombin activity.
- Pooling plasma increased thrombin generation threefold; heparin/coumarin use decreased it tenfold.
- Freezing and thawing plasma influenced thrombin generation, with 37°C thawing yielding the highest increase.
- Valid thrombin activity measurements require assessment during the ascending phase of the generation curve.
Conclusions:
- The described method allows for the quantification of thrombin activity in recalcified plasma.
- Factors such as plasma pooling, anticoagulant use, and storage conditions significantly impact thrombin generation.
- This assay can be utilized for diagnosing prothrombotic states by analyzing thrombin generation patterns.

