Pharm Sci. 2016;22(1):2-8.
doi: 10.15171/PS.2016.02

Scopus id: 84963614721
  Abstract View: 855
  PDF Download: 1238

Research Article

Development and Validation of A Spectrofluorimetric Determination of Calf Thymus DNA Using a Terbium-Danofloxacin Probe

Naser Soltani 1, Jamshid Manzoori 2, Mohammad Amjadi 3, Farzaneh Lotfipour 4, Abolghasem Jouyban 5,6 *

1 Drug Applied Research Center, Tabriz University o f Medical Sciences, Tabriz , Iran
2 Department of Chemistry, Tabriz Islamic Azad University, Tabriz, Iran
3 Department of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Iran
4 Hematology & Oncology Research Center and Faculty of Pharmacy, Tabriz University o f Medical Sciences, Tabriz , Iran
5 Pharmaceutical Analysis Research Center and Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran
6 Kimia Idea Pardaz Azarbayjan (KIPA) Science Based Company, Tabriz University o f Medical Sciences, Tabriz , Iran

Abstract

Background: Analysis of biomolecules is required in many biomedical research areas. A spectrofluorimetric method is proposed for determination of calf thymus DNA (ctDNA) based on the fluorescence enhancement of terbium-danofloxacin (Tb3+-Dano) in the presence of ctDNA. Methods: A probe with maximum excitation and emission wavelengths of 347 nm and 545 nm, respectively, was developed. The enhanced fluorescence intensity of Tb3+-Dano system was proportional to the concentration of ctDNA. The effective factors and the optimum conditions for the determination of ctDNA were studied. Under the optimum conditions of [Tris buffer]= 0.01 mol L-1 (pH 7.8), [ Tb3+]= 1×10-5 mol L-1 and [Dano]= 5×10-5 mol L-1, the maximum response was achieved. The developed method was evaluated in terms of accuracy, precision and limit of detection. Results: The linear concentration range for quantification of ctDNA was 36-3289 ng mL-1 and the detection limit (S/N=3) was 8 ng mL-1. The concentration of DNA extracted from Escherichia coli as an extracted sample was also determined using the developed probe. The concentration of DNA in extracted sample was determined using UV assay and developed method, the results were satisfactory. Conclusion: The proposed method is a simple, practical and relatively interference free method to follow up the concentrations of ctDNA.
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Submitted: 13 Aug 2015
Accepted: 10 Oct 2015
First published online: 30 Mar 2016
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