Acta Pharm. 49 (1999) 35–41

Detection of heparin during F VIII isolation using improved azure A method

BRANKA GEBAUER,^* KARMEN BRANOVIC,  MIRJANA ŠIMIC,  ANDJA TREŠCEC, 
and  BOJAN BENKO

¹Institute of Immunology,  Rockefeller Street 2,  10 000 Zagreb,  Croatia

Cryoprecipitate with heparin was the starting material in chromatographic isolation of factor VIII (F VIII). Heparin stabilizes F VIII and in that sense enhances the recovery of F VIII during isolation. The heparin concentration during different steps of F VIII isolation procedure was determined by modified azure A method. Azure A method is a rapid and simple spectrometric method for determination of heparin following the formation of soluble complex between heparin and azure A dye. Influence of different parameters (pH, heparin concentration, stability of the dye and heparin-dye complex) on azure A method has been established. Results presented in this report show that determination of heparin is achieved by simultaneous measuring absorbance of the complex at 500 nm and azure A dye absorbance decrease at 630 nm.

Keywords: F VIII, heparin, azure