Development and clinical evaluation of two chromogenic substrate methods for monitoring fondaparinux sodium.

This study was designed to develop methods for monitoring of the selective factor Xa inhibitor fondaparinux sodium (ARIXTRA) based on standard laboratory methods for the chromogenic determination of the anti-factor Xa activity of low molecular weight heparin. To examine the biologic activity of fondaparinux in comparison to its plasma concentration, 2 methods were investigated: 1 working with the addition of antithrombin (AT), the other without exogenous AT. Both methods showed a linear relationship of fondaparinux concentration and OD/min on a log-lin scale in the range from 0.1 to 2 microg/mL. Inter- and intra-assay variability was <6% in all cases. The results of spiked samples from patients on vitamin K antagonists (VKA) were in good agreement with both methods, and the determination of the fondaparinux concentration was not influenced by reduced levels of factor X in plasma caused by VKA-intake. Ex vivo samples from orthopedic patients (n=18) on prophylactic treatment with fondaparinux showed concentrations between 0.2 to 0.7 microg/mL 3 hours after s.c. injection. No significant differences were detected between both methods with these samples. The presented methods are suitable tools for monitoring of fondaparinux. The linear calibration curve in the range 0.1 to 2 microg/mL is suitable for determination of prophylactic and therapeutic application of fondaparinux. Both methods, with and without addition of AT, can be performed fully automated in clinical routine on an automated coagulation analyzer (STA coagulation analyzer). No significant differences were detected between both methods with these samples.

Univariate tolerance regions for fibrinogen, antithrombin III, protein C, protein S, plasminogen and alpha 2-antiplasmin in children using the new Automated Coagulation Laboratory (ACL) method.

To avoid misclassification of lowered or enhanced coagulation proteins in childhood the purpose of this study was to establish functional normal ranges for healthy children aged 6 months to 16 years. PT, aPTT, fibrinogen, antithrombin III, protein C, protein S, plasminogen and alpha 2-antiplasmin were tested using the new Automated Coagulation Laboratory (ACL) method. Values for PT, aPTT, fibrinogen, antithrombin III, plasminogen and alpha 2-antiplasmin were closely comparable in all children although we found a minimum range of 61% in children aged 8 to 16 years in plasma concentrations of alpha 2-antiplasmin. Children < 2.5 years showed reduced lower boundaries encompassing 95% of the population for protein C and protein S activity, although medians for protein S activity were similar in all children. The rapid and automatic determination of functional coagulation proteins using chromogenic substrates on the ACL 300 (25-50 microliters citrated plasma/test) renders the possibility to realize a screening program for inherited thrombotic syndromes in a routine laboratory.

Regression of deep vein thrombosis by i.v.-administration of a low molecular weight heparin--results of a pilot study.

Twenty-five patients with phlebographically confirmed deep vein thrombosis of the lower limb were treated with intravenous infusions of low molecular weight heparin for 7 to 29 days. The mean dosage was 15.2 +/- 3.0 Uanti-Xa (equivalent 7.6 +/- 1.5 U-aPTT). Phlebograms were taken before, during and after the treatment with low molecular weight heparin and evaluated using the score system of Marder. Nearly complete recanalization of the occluded veins was found in six (24%) patients, improvement of the Marder score of 60 to 90% was found in four patients and of 30 to 60% in seven patients, while eight patients remained unchanged. With an average dose of 15.2 I.U./kg/h the heptest was prolonged to 70 to 120 seconds while the aPTT-level did not significantly increase. tPA-antigen-levels increased significantly in most of the patients after the third day of treatment, while PAI-activity remained unchanged. A positive conclusion between the decrease of the Marder score and the duration of treatment was found. Thus the low molecular weight heparin used in this investigation proved to be effective and safe in treating deep vein thrombosis.