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Spectrophotometric bioassay method for urokinase
Ravindra V Badhe1, Ravindra B Chitamani, Mayur N Bhujbal
1Dr. D. Y. Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune-18, MS, India. badheravi@rediffmail.com
Introduction:
Urokinase is a potent plasminogen activator. Present Bio assay methods of Urokinase are very tedious. So a new simple spectrophotometric Bio assay method was developed to estimate thrombolytic activity of Urokinase.
Methods And Results:
This Bio assay is designed for the quantitative in vitro spectrophotometric determination of Urokinase activity by utilizing its thrombolytic activity to carry out the lysis of plasma clots. The initial concentration of the plasma clots was adjusted to 0.2+/-0.01 absorbance and the linear decrease in the absorbance by addition of Urokinase concentration from 200 to 1200 IU/ml at lambda(max) 530 nm was studied. The activity of sample Urokinase can be quantified by comparing the absorbance of sample Urokinase with Urokinase standard Bio assay calibration curve and predicted in IU. The r(2) value of standard calibration curve was found out to be 0.993. The repeatability of the Bio assay was studied by performing the experiment six times with fresh plasma samples and the results showed significant similarity.
Discussion:
We can conclude that the novel Bio assay method was found to be simple, economical, reproducible and accurate than the present Bio assay methods.
