Applicability of FTIR-ATR method to measure carbonyls in blood plasma after physical and mental stress

Introduction. Oxidative stress is a state of imbalance between the production of reactive oxygen species and antioxidant defenses. It results in the oxidation of all cellular elements and, to a large extent, proteins, causing inter alia the formation of carbonyl groups in their structures. The study focused on assessment of changes in the plasma protein-bound carbonyls in police horses after combat training and after rest and the applicability of infrared spectroscopy with a Fourier transform, utilizing the attenuated total reflectance (FTIR-ATR) in detecting plasma protein oxidation. Methods. We evaluated the influence of both the different concentrations of hydrogen peroxide and combat training on protein carbonylation in horse blood plasma. The oxidation of plasma proteins was assessed using a spectrophotometric method based on the carbonyl groups derivatization with 2,4-dinitrophenylhydrazine (DNPH). The measured values were correlated with the carbonyl groups concentrations determined by means of the FTIR-ATR method. Results. The linear correlation between the DNPH and FTIR-ATR methods was shown. The concentration of plasma protein-bound carbonyls significantly deceased in police horses after one-day rest when compared to the values measured directly after the combat training (a drop by 23%, pThis work was financed under the program of the Minister of Science and Higher Education “Strategy of Excellence - University of Research” in 2018-2019 project number 0019/SDU/2018/18 in the amount of PLN 700 000 and partly by the statutory funds of Wrocław University of Science and Technology. This work was created in cooperation with the Ministry of the Interior of the Slovak Republic, Department of Police Kynology and Hippology. Tomasz Walski was supported by the European Union’s Horizon 2020 Research and Innovation Programme, under the Marie Skłodowska-Curie Grant Agreement No 713690, and by the Science Foundation Ireland (SFI) and the European Regional Development Fund (Grant Number 13/RC/2073)