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Polarographic Determination of Genotoxic 2-Amino-9-fluorenone
Hájková, Andrea ; Vyskočil, Vlastimil (advisor) ; Dejmková, Hana (referee)
of Bachaleor Thesis The aim to this work was to find the optimum conditions for polarographic determination of genotoxic 2-amino-9-fluorenone (2-AFN) that can negatively affect the living and working environment. Foremost, the stability of 2-AFN stock solution in methanol was monitored using UV-VIS spectrophotometry and an optimal ratio of buffered aqueous and methanolic phase was found out for following polarographic measurements at a classical mercury dropping electrode (DME). The dependence of polarographic behavior of 2-AFN (of concentration 1×10-4 mol dm-3 ) on pH of analyzed solution was measured using DC tast polarography (DCTP) and differential pulse polarography (DPP) in the pH range 1.9 - 12.8, in the medium of Britton-Robinson (BR) buffer - methanol (9:1). The mechanism of polarographic reduction of electrochemically reducible oxo group at 2-AFN has been proposed on the basis of thus observed behavior. The optimum medium of BR buffer pH 4.0 - methanol (9:1) has been chosen for both polarographic techniques. The concentration range from 1×10-6 to 1×10-4 mol dm-3 was measured using DCTP at DME and from 1×10-7 to 1×10-4 mol dm-3 using DPP at DME. Reached limits of quantification (LQs) were 5×10-7 mol dm-3 and 1×10-7 mol dm-3 for DCTP at DME and DPP at DME, respectively. For comparison, the...
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Novel Approaches in Electrochemical Determination of Xenobiotic Compounds and in Study of Their Interaction with DNA
Hájková, Andrea ; Vyskočil, Vlastimil (advisor) ; Trnková, Libuše (referee) ; Labuda, Ján (referee)
Presented Ph.D. Thesis is focused on the development of analytical methods applicable for determination of selected xenobiotic compounds and for monitoring DNA damage they can induce. The main attention has been paid to the development and testing of non-toxic electrode materials for preparation of miniaturized electrochemical devices and novel electrochemical DNA biosensors. 2-Aminofluoren-9-one (2-AFN) was selected as a model environmental pollutant, which belongs to the group of hazardous genotoxic substances. Its carcinogenic and mutagenic effects may represent a risk to living and working environment. 2-AFN has one oxo group, where the cathodic reduction occurs, and one amino group, where the anodic oxidation occurs. The voltammetric behavior of 2-AFN in the negative potential region was investigated at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE) representing a non-toxic and more mechanically robust alternative to mercury electrodes. This working electrode was subsequently used for the development of a newly designed miniaturized electrode system (MES), which has many benefits as the possibility of simple field measurements, easy portability, and the measurement in sample volume 100 µL. Moreover, a glassy carbon electrode (GCE) was used for further investigation of...
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Novel Approaches in Electrochemical Determination of Xenobiotic Compounds and in Study of Their Interaction with DNA
Hájková, Andrea
Presented Ph.D. Thesis is focused on the development of analytical methods applicable for determination of selected xenobiotic compounds and for monitoring DNA damage they can induce. The main attention has been paid to the development and testing of non-toxic electrode materials for preparation of miniaturized electrochemical devices and novel electrochemical DNA biosensors. 2-Aminofluoren-9-one (2-AFN) was selected as a model environmental pollutant, which belongs to the group of hazardous genotoxic substances. Its carcinogenic and mutagenic effects may represent a risk to living and working environment. 2-AFN has one oxo group, where the cathodic reduction occurs, and one amino group, where the anodic oxidation occurs. The voltammetric behavior of 2-AFN in the negative potential region was investigated at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE) representing a non-toxic and more mechanically robust alternative to mercury electrodes. This working electrode was subsequently used for the development of a newly designed miniaturized electrode system (MES), which has many benefits as the possibility of simple field measurements, easy portability, and the measurement in sample volume 100 µL. Moreover, a glassy carbon electrode (GCE) was used for further investigation of...
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Novel Approaches in Electrochemical Determination of Xenobiotic Compounds and in Study of Their Interaction with DNA
Hájková, Andrea ; Vyskočil, Vlastimil (advisor) ; Trnková, Libuše (referee) ; Labuda, Ján (referee)
Presented Ph.D. Thesis is focused on the development of analytical methods applicable for determination of selected xenobiotic compounds and for monitoring DNA damage they can induce. The main attention has been paid to the development and testing of non-toxic electrode materials for preparation of miniaturized electrochemical devices and novel electrochemical DNA biosensors. 2-Aminofluoren-9-one (2-AFN) was selected as a model environmental pollutant, which belongs to the group of hazardous genotoxic substances. Its carcinogenic and mutagenic effects may represent a risk to living and working environment. 2-AFN has one oxo group, where the cathodic reduction occurs, and one amino group, where the anodic oxidation occurs. The voltammetric behavior of 2-AFN in the negative potential region was investigated at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE) representing a non-toxic and more mechanically robust alternative to mercury electrodes. This working electrode was subsequently used for the development of a newly designed miniaturized electrode system (MES), which has many benefits as the possibility of simple field measurements, easy portability, and the measurement in sample volume 100 µL. Moreover, a glassy carbon electrode (GCE) was used for further investigation of...
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Neutral genetic variability and structure in chamois populations in Slovakia
Hájková, Andrea ; Zima, Jan (advisor) ; Kotlík, Petr (referee)
The Tatra chamois (Rupicapra rupicapra tatrica), an endemic mountain ungulate, occurs in the High Tatra Mts. (northern Slovakia and southern Poland). In the second part of the 20th century several chamois introductions occurred in Slovakia: Tatra chamois (from the High Tatra Mts.) were introduced into the Low Tatra Mts., while Alpine chamois (R. r. rupicapra) into the Veľká Fatra and Slovenský raj Mts. The High Tatra Mts. population underwent several population declines (bottlenecks) and all other Slovak populations were founded from only a few individuals (founder effect). Moreover, because the Low Tatra, Veľká Fatra and Slovenský raj are neighbouring mountain ranges, there is a potential risk of migration and hybridisation between the subspecies. Using 18 microsatellite loci, we studied neutral genetic variability, structure and potential hybridisation in chamois populations in Slovakia. The study is based on 193 samples: 95 tissue and 5 blood samples, and 88 faecal and 5 hair samples. Low genetic variability was found in all populations, the lowest one in High Tatra Mts. population. High values of fixation index, the number of private alleles, and factorial correspondence analysis indicated strong differentiation between the studied populations. Bayesian clustering divided Slovak chamois...
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Voltammetric Determination of Trace Amounts of Genotoxic 2-Aminofluoren-9-one
Hájková, Andrea ; Vyskočil, Vlastimil (advisor) ; Navrátil, Tomáš (referee)
of the Diploma Thesis Presented Diploma Thesis is focused on the electrochemical determination of genotoxic 2-aminofluoren-9-one (2-AFN), which belongs to derivatives of polycyclic aromatic hydrocarbons. Its possible carcinogenic and mutagenic effects may represent a risk to living and working environment. Optimal conditions for the voltammetric determination of 2-AFN were found in an appropriate ratio of buffered aqueous and methanolic phase. In the methanol - Britton- Robinson (BR) buffer (1:9) medium, voltammetric behavior of 2-AFN at a concentration of 1×10-4 mol/l was investigated in a dependence on the pH of the solution in the range from 2.0 to 13.0. For following measurements of concentration dependences, the optimal medium was chosen to be methanol - BR buffer of pH 4.0 (1:9). The achieved limits of quantification (LQ) were 2×10-7 mol/l for DC voltammetry (DCV) and 1×10-7 mol/l for differential pulse voltammetry (DPV) at a hanging mercury drop electrode (HMDE). In the medium of an acetate buffer of pH 4.0, the LQ attained using adsorptive stripping differential pulse voltammetry (AdSDPV) at the HMDE under optimal conditions for the accumulation of the analyte (Eacc = +100 mV and tacc = 120 s) was 4×10-9 mol/l of 2-AFN. For the voltammetric determination of 2-AFN at a mercury meniscus...
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Novel Approaches in Electrochemical Determination of Xenobiotic Compounds and in Study of Their Interaction with DNA
Hájková, Andrea
Presented Ph.D. Thesis is focused on the development of analytical methods applicable for determination of selected xenobiotic compounds and for monitoring DNA damage they can induce. The main attention has been paid to the development and testing of non-toxic electrode materials for preparation of miniaturized electrochemical devices and novel electrochemical DNA biosensors. 2-Aminofluoren-9-one (2-AFN) was selected as a model environmental pollutant, which belongs to the group of hazardous genotoxic substances. Its carcinogenic and mutagenic effects may represent a risk to living and working environment. 2-AFN has one oxo group, where the cathodic reduction occurs, and one amino group, where the anodic oxidation occurs. The voltammetric behavior of 2-AFN in the negative potential region was investigated at a mercury meniscus modified silver solid amalgam electrode (m-AgSAE) representing a non-toxic and more mechanically robust alternative to mercury electrodes. This working electrode was subsequently used for the development of a newly designed miniaturized electrode system (MES), which has many benefits as the possibility of simple field measurements, easy portability, and the measurement in sample volume 100 µL. Moreover, a glassy carbon electrode (GCE) was used for further investigation of...
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Polarographic Determination of Genotoxic 2-Amino-9-fluorenone
Hájková, Andrea ; Dejmková, Hana (referee) ; Vyskočil, Vlastimil (advisor)
of Bachaleor Thesis The aim to this work was to find the optimum conditions for polarographic determination of genotoxic 2-amino-9-fluorenone (2-AFN) that can negatively affect the living and working environment. Foremost, the stability of 2-AFN stock solution in methanol was monitored using UV-VIS spectrophotometry and an optimal ratio of buffered aqueous and methanolic phase was found out for following polarographic measurements at a classical mercury dropping electrode (DME). The dependence of polarographic behavior of 2-AFN (of concentration 1×10-4 mol dm-3 ) on pH of analyzed solution was measured using DC tast polarography (DCTP) and differential pulse polarography (DPP) in the pH range 1.9 - 12.8, in the medium of Britton-Robinson (BR) buffer - methanol (9:1). The mechanism of polarographic reduction of electrochemically reducible oxo group at 2-AFN has been proposed on the basis of thus observed behavior. The optimum medium of BR buffer pH 4.0 - methanol (9:1) has been chosen for both polarographic techniques. The concentration range from 1×10-6 to 1×10-4 mol dm-3 was measured using DCTP at DME and from 1×10-7 to 1×10-4 mol dm-3 using DPP at DME. Reached limits of quantification (LQs) were 5×10-7 mol dm-3 and 1×10-7 mol dm-3 for DCTP at DME and DPP at DME, respectively. For comparison, the...
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