title: Validation of the anti-factor IIa assay and potency assessment of enoxaparin in pharmaceutical formulations.
abstract: The low molecular weight heparin enoxaparin sodium is used clinically for the prevention and treatment of venous and arterial thrombosis. An anti-factor IIa assay was applied and validated for the potency evaluation of the pharmaceutical formulations. Investigating the parameters of range, linearity (r(2)=0.9912), precision, accuracy and robustness, the biological assay incorporated a chromogenic end-point and detection at 405 nm. The method yielded good results with a detection limit of 0.01 IU/ml and a quantitation limit of 0.03 IU/ml. Sodium enoxaparin pharmaceutical products were evaluated by the anti-factor Xa assay and the anti-factor IIa assay giving potencies between 97.18% and 106.50%, with an anti-factor Xa/anti-factor IIa ratio between 3.9 and 4.2. The results demonstrated the validity of the anti-factor IIa assay that together with the anti-factor Xa are useful methodologies for the routine quality control of enoxaparin in pharmaceutical products.
authors.affiliation: Department of Industrial Pharmacy, Health Science Centre, Federal University of Santa Maria, 97.1105-900-Santa Maria-RS, Brazil. sdalmora@ccs.ufsm.br <sdalmora@ccs.ufsm.br>
authors.forename: Sérgio Luiz
authors.initials: SL
authors.lastname: Dalmora
authors.forename: Liberato Brum
authors.initials: LB
authors.lastname: Junior
authors.forename: Silvana Ferreira
authors.initials: SF
authors.lastname: Vaccari
authors.forename: Marcio
authors.initials: M
authors.lastname: Fronza
authors.forename: Paulo
authors.initials: P
authors.lastname: Renato de Oliveira
authors.forename: Clarice Madalena Bueno
authors.initials: CM
authors.lastname: Rolim
mesh_terms: D000925:Anticoagulants / Q000627:therapeutic use; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D017984:Enoxaparin / Q000032:analysis* / Q000494:pharmacology; D005343:Fibrinolytic Agents / Q000627:therapeutic use; D006495:Heparin, Low-Molecular-Weight / Q000627:therapeutic use; D011516:Prothrombin / Q000037:antagonists & inhibitors*; D015203:Reproducibility of Results; D013927:Thrombosis / Q000209:etiology / Q000517:prevention & control
chemical_list: D000925:Anticoagulants; D017984:Enoxaparin; D005343:Fibrinolytic Agents; D006495:Heparin, Low-Molecular-Weight; D011516:Prothrombin; C025337:Factor IIa