Novel, versatile capillary electrophoresis instrument with laser induced fluorescence for analysis
of various lipid peroxidation biomarkers.

Júlia Lačná^1,2, František Foret^1, Petr Kubáň^1,2


^1 Department of Bioanalytical instrumentation, CEITEC MU, Veveří 97, 602 00 Brno, Czech Republic

^2 Department of Chemistry, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic,

Contact information: Júlia Lačná, juliahod@gmail.com


When polyunsaturated fatty acids are attacked by reactive oxygen species, the oxidation products
such as malondialdehyde (MDA), 4-hydroxynonenal, and dienals are formed. Malondialdehyde (MDA) is
the most frequently studied biomarker of lipid peroxidation. The common method for analysis of MDA
is by fluorescence using the thiobarbituric acid (TBA) assay. The resulting product is highly
fluorescent (excitation 532 nm, emission 550 nm) and negatively charged. Its negative charge allows
an efficient separation and detection by CE with laser induced fluorescence (LIF) that is an ideal
combination for the analysis of low concentrations of analytes in small volumes of biological
samples. In this work, we have built a versatile CE-LIF system that allows easy exchange of
different laser modules for sensitive detection. A 532 nm green laser module was incorporated in
the system for sensitive determination of MDA. The reaction conditions of MDA with TBA was
optimized, including reaction time, reaction temperature, ratio of reagent to analyte and pH of
reaction solution. The separation conditions in the counter-electroosmotic mode were optimized for
sensitive MDA analysis. Other oxidative stress markers can potentially also be separated and
detected.


Acknowledgments

The authors acknowledge the financial support from the Grant Agency of the Czech Republic (Grant
No. P206/13/21919S). Part of the work was realized in CEITEC - Central European Institute of
Technology with research infrastructure supported by the project CZ.1.05/1.1.00/02.0068 financed
from European Regional Development Fund.