Validation of a Chromogenic Substrate Method for Biological Activity Quantification of Streptokinase

by Ana Aguilera, Lázara Muñoz, Yilian Bermúdez, Danae Arias, Yamila Martínez, Gerardo García, Luciano Hernández, Eduardo Martínez, and Rodolfo Valdés
Volume 13, Issue 3 (Fall 2014)

The use of thrombolytic agent, streptokinase (SK), has been adapted to treat patients with the medical condition, hemorrhoids, in a new pharmaceutical formulation. Part of the development process included studies for measuring SK biological activity in the suppository product. Thus, the main objective in this study was in adapting and validating an established chromogenic substrate method for biological activity quantification of SK extracted from suppositories. By using several solutions and methods described in this paper, results revealed 103.82–124.99% of SK biological activity recovery and high molecular integrity. The chromogenic substrate method was specific for SK, and linear from 200– 600 IU mL-1 (R2 = 0.995 ± 0.002) with a variation coefficient of less than 7.11% and 12.62% for repeatability and reproducibility experiments, respectively. SK biological activity values estimated by the chromogenic substrate method were comparable with those estimated by the clot lysis method used as the reference. We have concluded that the method validated in this study was specific, accurate, and precise for quantifying biological activity of SK extracted from a suppository formulation intended to treat patients with acute hemorrhoids.

Aguilera A, Muñoz L, Bermúdez Y, Arias D, Martínez Y, García G, Hernández L, Martínez E, Valdés R. Validation of a chromogenic substrate method for biological activity quantification of streptokinase. BioProcess J, 2014; 13(3): 49–59.

Posted online October 13, 2014.