Bioimpacts. 2018;8(1): 31-38.
doi: 10.15171/bi.2018.05
PMID: 29713600
PMCID: PMC5915706
Scopus ID: 85044265885
WOSID: WOS:000429104100005
  Abstract View: 1645
  PDF Download: 1125
  Full Text View: 676

Original Research

Spectrophotometric analysis of thrombolytic activity: SATA assay

Masumeh Zamanlu, Morteza Eskandani * , Reza Mohammadian, Nazila Entekhabi, Mohammad Rafi, Mehdi Farhoudi *


Introduction: Measurement of thrombolytic activity is crucial for research and development of novel thrombolytics. It is a key factor in the assessment of the effectiveness of conventionally used thrombolytic therapies in the clinic. Previous methods used for the assessment of thrombolytic activity are often associated with some drawbacks such as being costly, time-consuming, complex with low accuracy. Here, we introduce a simple, economic, relatively accurate and fast method of spectrophotometric analysis of thrombolytic activity (SATA) assay, standardized by tissue plasminogen activator (tPA), which can quantitatively measure in vitro thrombolytic activity.

Methods: Blood clots were formed, uniformly, by mixing citrated whole blood with partial thromboplastin time (PTT) reagent, together with calcium chloride. Then, designated concentrations of tPA were added to the samples, and the released red blood cells from each clot were quantified using spectrophotometry (λmax=405nm) as an indicator of thrombolytic activity. The accuracy of the method was tested by assessment of dose-responsibility against R2 value obtained by linear equation and measurement of the limit of detection (LOD) and limit of quantification (LOQ). The SATA assay was validated in comparison with some currently used techniques.

Results: A linear relationship was obtained between different concentrations of tPA versus the spectrophotometric absorbance of the related dilutions of lysed clots, at λmax=405nm. Calculated R2 values were greater than 0.9; with LOD of 0.90 µg/mL of tPA (436.50IU) and LOQ of 2.99 µg/mL of tPA (1450.15IU).

Conclusions: Conclusively, the SATA assay is a very simple quantitative method with repeatable and reproducible results for estimating the potency of an unknown thrombolytic agent, and calculating the activity as delicate as 1 µg/mL of tPA (485 IU/mL of thrombolytic dose). 

Keywords: Clot lysis, Thrombolytic agent, In vitro thrombolysis, Thrombolytic therapy, Fibrinolysis
First Name
Last Name
Email Address
Security code

Abstract View: 1645

Your browser does not support the canvas element.

PDF Download: 1125

Your browser does not support the canvas element.

Full Text View: 676

Your browser does not support the canvas element.

Submitted: 31 Jul 2017
Revision: 28 Oct 2017
Accepted: 31 Oct 2017
ePublished: 01 Nov 2017
EndNote EndNote

(Enw Format - Win & Mac)

BibTeX BibTeX

(Bib Format - Win & Mac)

Bookends Bookends

(Ris Format - Mac only)

EasyBib EasyBib

(Ris Format - Win & Mac)

Medlars Medlars

(Txt Format - Win & Mac)

Mendeley Web Mendeley Web
Mendeley Mendeley

(Ris Format - Win & Mac)

Papers Papers

(Ris Format - Win & Mac)

ProCite ProCite

(Ris Format - Win & Mac)

Reference Manager Reference Manager

(Ris Format - Win only)

Refworks Refworks

(Refworks Format - Win & Mac)

Zotero Zotero

(Ris Format - Firefox Plugin)