Validation of the antifactor Xa and antifactor IIa assays for the potency assessment of nadroparin in pharmaceutical formulations

• Published in: Journal of AOAC International 2006, Vol.89 (1), p.58-64
• Main Authors: DALMORA, Sérgio Luiz, SILVA SANGOI, Maximiliano Da, BARTH, Thiago, BRUM, Liberato, FERREIRA VACCARI, Silvana
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Calibration; Cattle; Chemical properties; Chemistry Techniques, Analytical - methods; Chemistry, Pharmaceutical - methods; Drugs; Evaluation; Factor Xa - analysis; Factor Xa - chemistry; Food industries; Fundamental and applied biological sciences. Psychology; Health aspects; Heparin; Humans; Nadroparin - analysis; Nadroparin - chemistry; Product/Service Evaluations; Prothrombin - analysis; Prothrombin - chemistry; Quality Control; Reproducibility of Results; Thrombosis - prevention & control; Time Factors; Venous Thrombosis - prevention & control
• ISSN: 1060-3271; 1944-7922
• DOI: 10.1093/jaoac/89.1.58

The low-molecular weight heparin nadroparin calcium is used clinically for the prevention and treatment of venous and arterial thrombosis. The antifactor Xa and antifactor IIa assays were validated by investigating the parameters of range, linearity (r2 = 0.9905 and r2 = 0.9914, respectively) precision, accuracy, and robustness. The 2 methods incorporated a chromogenic endpoint and detection at 405 nm, yielding good results with detection limits of 0.004 and 0.01 IU/mL and quantitation limits of 0.01 and 0.03 IU/mL, respectively, for the antifactor Xa and antifactor IIa assays. Nadroparin calcium pharmaceutical products were evaluated by the antifactor Xa assay and the antifactor IIa assay, giving potencies between 93.86 and 109.88%, with an antifactor Xa/antifactor IIa ratio between 3.2 and 3.7. The results demonstrated the validity of the assays that are useful methodologies for the routine quality control of nadroparin in pharmaceutical formulations.