Project PN-III-P1-1_1-TE-2021-0655
New chromatographic approaches for evaluation of the antioxidant/prooxidant profile of redox-active drugs

Scientific results obtained during the implementation period of the project with the title:
New chromatographic approaches to evaluate the antioxidant/prooxidant profile of
redox-active drugs
Project Acronim: Redox-TLC-IA
Project code: PN-III-P1-1.1-TE-2021-0655
Period of implementation: 13.05.2022 - 12.05.2024
➢ Within this project, new methods were developed and validated for the evaluation of antioxidant potential of drugs using DPPH•and ABTS•+ radicals and micro-HPTLC and HPTLC chromatographic techniques coupled with advanced image processing and analysis methods (micro HPTLC- IA, HPTLC-IA).
➢ Compared to the methods found in the literature, the new micro-HPTLC-IA and HPTLC-IA methods developed in this project allow the rapid determination of the antiradical activity of drugs against DPPH•and ABTS•+ radicals and the evaluation of two parameters that include total antioxidant potential (TAP) and antioxidant potential after interaction of drugs with different substrates that simulates the interactions with the physiological environment. It is important to note that for drugs structurally related to catecholamines, the determination of antiradical activity against the two radicals is very important because these drugs may have different interaction mechanisms with free radicals depending on the circumstances of the physiological environment.
➢ Using the developed chromatographic methods, the antiradical profile for a set of adrenergic drugs was determined in simulated physiological conditions using reversed phase silica gel plates RP-8, RP 18, CN and Diol. Based on the obtained results, the antioxidant potential of these drugs was evaluated after their interaction with the studied stationary phases.
➢ The synergistic/antagonistic effect of mixtures of adrenergic drugs with adrenaline was evaluated based on their reaction with DPPH•and ABTS•+ radical respectively and using the HPTLC-IA method developed in this project. According to the obtained results, drugs that have an antagonistic, additive, synergistic or possible pro-oxidant effect in combination with adrenaline were highlighted.
➢ New sensitive methods were developed for rapid detection and determination of adrenergic drugs banned in sports competitions using DPPH•and ABTS•+ radicals as sensitive detection reagents and the HPTLC-IA method developed within this project.
➢ The dissemination of the obtained results was carried out both nationally and internationally by participation to a number of four scientific conferences, the publication of two scientific papers in ISI rated journals (Studia Universitatis Babes-Bolyai, Chemia and Journal of Liquid Chromatography & Related Technologies) and submitting a scientific paper for publication in Journal of Separation Science.
Quantifiable results achieved during the project implementation period
➢ Creating/updating the project web page that includes the summary, objectives and stages of the project implementation, the project team and results achieved during the project implementation. Web page address: https://dorinaresearch.lm.r.appspot.com/
➢Participation to national and international conferences
1. Dorina Casoni, Alexandra-Gabriela Kiraly, Simona Codruța Cobzac, New method for rapid identification and determination of selective adrenergic drugs using the micro-TLC approach, Conferința Națională de Chimie, Ediția XXXVI, Călimănești-Căciulata, 04- 07 octombrie 2022 (Poster - Anexa 1).
2. Dorina Casoni, Ileana-Maria Simion, Melinda Carmen Szasz, New chromatographic approach for rapid evaluation of antioxidant potential of adrenergic dugs, 49th International Conference of SSCHE, Tatranske Matliare, Slovakia, 15 - 18 May, 2023 (Poster – Anexa 2).
3. Dorina Casoni, Melinda Carmen Szasz, Ileana-Maria Simion, Sensitive HPTLC method for simultaneous determination of adrenergic drugs prohibited in sport, 27th International Symposium on Separation Sciences (ISSS 2023), September 24th-27th, 2023, Cluj Napoca, Romania (Prezentare orală- Anexa 3).
4. Dorina Casoni, Larisa Silveșan, Evaluation of substrate-interaction effect on the antioxidant potential of adrenergic drugs using thin-layer chromatography with image analysis method, 50th International conference of Slovak Society of Chemical Engineering – SSCHE 2024, Tatranské Matliare, Slovakia, 20-24 May, 2024 (Poster – Anexa 4).
➢Scientific papers published in ISI journals / sent for publication
1. Casoni D., Simion I. M., Cobzac S.C.A., Kiraly A.G., Development of a new micro-HPTLC protocol for total antioxidant potential determination of redox-active drugs, Studia Universitatis Babes-Bolyai, Chemia 67(4) (2022) 235-247 DOI: 10.24193/subbchem.2022.4.15 (Q4, IF = 0.231)
2. Szasz M., Casoni D., Cimpoiu C., Study on binding affinity profile, bioactivity, and lipophilicity of selected antipsychotic drugs, Journal of Liquid Chromatography & Related Technologies (2023) 1-6. DOI10.1080/10826076.2023.2216780 (Q4, IF = 1.3))
3. Dorina Casoni, Melinda Carmen Szasz, Eda Buker, Simona Codruța Aurora Cobzac, Ileana Maria Simion, New method for rapid detection and simultaneous determination of prohibited adrenergic drugs in sports using thin layer chromatography and image processing, Journal of Separation Science, trimis spre publicare (Manuscript ID: jssc.202400268) (Q2. IF=3.37).
The estimated impact of the obtained results
Considering the innovation and originality of the micro-HPTLC-IA and respectively HPTLC-IA methods developed and validated within this project, it is expected that the obtained results to have an important impact in the field of analytical chemistry, pharmacology and also in the scientific field.
Regarding the field of analytical chemistry, by the developed project new protocols were validated for the rapid evaluation of the antioxidant potential of compounds using thin-layer chromatography coupled with image analysis techniques and DPPH•și ABTS*+ radicals.
➢ Both the micro-HPTLC-DPPH• and the micro-HPTLC-ABTS*+ protocol developed within the project can be used in the rapid evaluation of the antioxidant potential of redox-active compounds. Compared to the reference methods developed in the literature up to the present, these new methods allow the separation of the compounds resulting from the reaction with DPPH•and ABTS*+ radicals and thus allows a more precise evaluation of the total antioxidant potential of the compounds especially if the reaction with the radical leads to the formation of colored compounds that can affect determinations by reference methods.
➢ The HPTLC-IA method developed using for the first time an innovative approach based on the use of modified silica gel chromatographic plates as interaction substrates simulating biological membranes, can be used in the rapid estimation of the variation of the antioxidant potential of compounds after their interaction with different constituents from physiological environment.
Regarding the pharmaceutical area, the results obtained in this project provide for the first time new information related to the antiradical profile of a wide set of adrenergic drugs, the variation of their antioxidant potential after the interaction with possible constituents of the physiological environment and the synergistic/antagonistic activity of these drugs in combination with catecholamines (adrenaline).
➢ The micro-HPTLC-DPPH•and respectively micro-HPTLC-ABTS*+ protocol developed within the project can be used in the rapid evaluation of the total antioxidant potential of redox-active drugs with activity from pharmaceutical and medical point of view.
➢ The HPTLC-IA method developed within the project can be used to estimate the variation of the antioxidant potential of some drugs of pharmaceutical interest under conditions that simulate the physiological environment as well as the possible interaction with constituents from the physiological environment.
➢ The HPTLC-IA method developed for the separation, identification and determination of adrenergic drugs using sensitive detection techniques, can be implemented in the rapid identification and determination of prohibited adrenergic drugs in sports competitions.
From a scientific point of view, the results presented within a number of four international conferences and published in three scientific articles present important aspects that can be used as starting points in further research related to the field of chromatography and the evaluation of antioxidant activity of redox-active compounds.
➢ The results disseminated during the project implementation period provide useful information related to the: use of DPPH•and ABTS*+ radicals in chromatographic determinations; image processing/analysis techniques applied for sensitive detection with these radicals; use of the chromatographic plates as substrates that simulate the interaction of compounds with constituents of biological environment allowing the estimation of the effect of these interactions on the pharmacokinetic activity of drugs.