rd
23 Congress of the International Union for Biochemistry and Molecular Biology
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44 Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology
th
th
Foz do Iguaçu, PR, Brazil, August 24 to 28 , 2015
DEVELOPMENT OF METHODS FOR ANALYSIS OF DOPAMINE IN DIFFERENT
COLUMNS BY HPLC-DAD
GAYER, M.C.1; SOARES, J.J.2; VARGAS, L.S.3; MELLO-CARPES, P.B.3; DENARDIN, E.L.G.2;
1
ROEHRS, R.
1
Grupo Interdisciplinar de Pesquisa em Práticas de Ensino (GIPPE), Universidade Federal do Pampa
(UNIPAMPA), Uruguaiana, Rio Grande do Sul, Brasil.
2
Laboratório de Estudos Físico-Químicos e Produtos Naturais (LEFQPN), Universidade Federal do
Pampa (UNIPAMPA), Uruguaiana, Rio Grande do Sul, Brasil.
3
Grupo de Pesquisa em Fisiologia (GPFis), Universidade Federal do Pampa (UNIPAMPA), Uruguaiana,
Rio Grande do Sul, Brasil.
Introduction and Objectives: Dopamine is an important neurotransmitter that acts on
memory and movement processes. Some diseases such as Parkinson's disease and
schizophrenia are associated with changes in the levels of this neurotransmitter.
Studies with Wistar rats and Drosophila melanogaster are widely used to understand
the mechanisms involved in these diseases and the dopamine measurements are made
through HPLC analysis. From this, the development and the validation of a method is
necessary so that the analyzes are reliable. Thus, our objective was to evaluate two
chromatographic columns for HPLC-DAD, one Kinetex Hillic column and a Synergi
Fusion-RP column. Materials and Methods: The methods developed are consisting by
mobile phase of acetonitrile/water pH3 in the proportions of 94/06 (v:v) and flow 0.8
ml.min-1 for Hillic column and 25/75 (v:v) and flow of 1 ml.min-1 for Synergi. To assess
the applicability of each column in dopamine analysis were injected samples of brains of
rats and head of flies. In both methods, the analysis times were 10 minutes with
injection of 20 µL of sample, data acquisition at 198 ƞm and the range of detection and
quantification of 0.1 mg.L-1 at 10 mg.L-1. Results and Conclusions: Evaluation of the
methods happened with the analysis of the analytical curves for each column, in which
R2 = 0.9999 values were obtained for both columns. For Hillic and Synergi columns
there was obtained the equations y=962,35x-73,994 and y=628,21x-28,775,
respectively. The results show that the methods been effective in separating dopamine
of the other constituents of the sample, evaluating concentrations starting from 0.1
mg.L-1. The two columns can be used efficiently for the quantification of dopamine in
samples without complex processes of sample preparation and present themselves as
reliable alternatives for research with this purpose.
Acknowledgements:
Key Words: Dopamine, Chromatography, Development of Methods.
Brazilian Society for Biochemistry and