Method for simultaneously determining multiple coagulation proteases

摘要

The present invention relates to a chromogenic method for simultaneously determining the activity of multiple coagulation proteases or for simultaneously determining the inhibition of multiple coagulation proteases in a single test reaction. For this purpose, use is made of two chromogenic substrates which have different absorption maxima and whose color signals can be separated spectrally.

主权项

1. A method for determining anti-thrombin and anti-factor Xa anticoagulant concentrations in a test sample which contains human plasma and which is suspected of containing an anti-thrombin anticoagulant, an anti-factor Xa anticoagulant, or a combination thereof, the method comprising:
performing a single test reaction by mixing the test sample which contains the human plasma and which is suspected of containing the anti-thrombin anticoagulant, the anti-factor Xa anticoagulant, or the combination thereof, with selected substances including (a) a defined amount of thrombin, (b) a defined amount of factor Xa, (c) a first chromogenic substrate specific for thrombin, and (d) a second chromogenic substrate specific for factor Xa, wherein a chromophore of the first chromogenic substrate and a chromophore of the second chromogenic substrate have respective absorption maximums that differ by at least 100 nm; photometrically determining absorption changes in the single test reaction over time; comparing the photometrically determined absorption changes in the single test reaction with photometrically determined absorption changes occurring in a control test reaction between an anticoagulant-free control sample and the selected substances; determining inhibition of the absorption changes caused by the anti-thrombin and/or anti-factor Xa anticoagulants in the test sample based on the comparison of the photometrically determined absorption changes; and determining the anti-thrombin and anti-factor Xa anticoagulant concentrations in the test sample based on the inhibition of the absorption changes in the single test reaction.