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Application of modern separation techniques in the analysis of biological material
Červinková, Barbora ; Solich, Petr (advisor) ; Mikuš, Peter (referee) ; Štěrbová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Department of Analytical Chemistry Candidate Mgr. Barbora Červinková Supervisor prof. RNDr. Petr Solich, CSc. Title of Doctoral Thesis Application of modern separation techniques in the analysis of biological material The presented dissertation is focused on development, optimization and validation of new extraction procedures and chromatographic methods for determination of vitamins A and E, inflammation biomarkers, DNA and RNA oxidative stress products in biological matrices. In the theoretical part of this work selected analytes and possibility of their determination in biological materials are described. Moreover, modern trends in liquid chromatography and bioanalytical validation guidelines are included. In this part, special attention is focused on sample handling, transport and pre- treatment. The second part is focused on published methods and divided into two sections. The first section comments published developed analytical methods. As first method, determination of lipophilic vitamins in human serum by miniaturized liquid-liquid extraction with subsequent separation on fluorinated stationary phase using UHPLC system with fluorescent detection in 4 min is discussed. Thereafter, determination of creatinine, DNA and...
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Genetic polymorphism in the HFE gene in the Czech population
Červinková, Barbora ; Beránek, Martin (advisor) ; Šimůnek, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Červinková Supervisor: Doc. PharmDr. Martin Beránek, Ph.D. Title of diploma work: Genetic polymorphism of the HFE gene in Czech population Outline: The main objective of the thesis was to map the incidence of the three mutations (C282Y, H63D, S65C) in the HFE gene in the Czech population. Consequently, the obtained results were compared with the reported occurrence in the world. Methods: The work involved the isolation of DNA from buccal swabs obtained from 167 donors (65 men, 102 women; average age for men was 31, for women 28). Isolated DNA strings were amplified by PCR methods using 5'-CAG ATC CTC ATC TCA CTG-3' and 5'-CTG GAT AAC CTT GGC TGT ACC CCC-3' primers for C282Y mutation, 5'-GCC ACA TCT GGC TTG AAA TT-3' and 5'-ACA TGG TTA AGG CCT GTT GC GCC ACA- 3' primers for H63D and S65C mutations. DNA samples were treated with restriction enzymes Rsa I, Bcl I and Hinf I, for C282Y, H63D and S65C mutations, respectively. Finally, the restriction fragments were separated by gel electrophoresis (2 % agarose gel). Results: C282Y mutation was present in 0 (0 %) and 19 (11,38 %) samples as homozygote or heterozygote, respectively. Moreover, C282Y mutation was present in 3 (1,80 %)...
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Application of modern separation techniques in the analysis of biological material
Červinková, Barbora ; Solich, Petr (advisor) ; Mikuš, Peter (referee) ; Štěrbová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Department of Analytical Chemistry Candidate Mgr. Barbora Červinková Supervisor prof. RNDr. Petr Solich, CSc. Title of Doctoral Thesis Application of modern separation techniques in the analysis of biological material The presented dissertation is focused on development, optimization and validation of new extraction procedures and chromatographic methods for determination of vitamins A and E, inflammation biomarkers, DNA and RNA oxidative stress products in biological matrices. In the theoretical part of this work selected analytes and possibility of their determination in biological materials are described. Moreover, modern trends in liquid chromatography and bioanalytical validation guidelines are included. In this part, special attention is focused on sample handling, transport and pre- treatment. The second part is focused on published methods and divided into two sections. The first section comments published developed analytical methods. As first method, determination of lipophilic vitamins in human serum by miniaturized liquid-liquid extraction with subsequent separation on fluorinated stationary phase using UHPLC system with fluorescent detection in 4 min is discussed. Thereafter, determination of creatinine, DNA and...
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Women awareness of breast cancer prevention
Červinková, Barbora ; Di Cara, Veronika (advisor) ; Kulhavá, Miluše (referee)
Prevention of breast cancer using the form of mammographic screening is one of the most important factors involved in early diagnosis and reduction of mortality with this type of cancer, which is the most frequently occurring cancer in women in the Czech Republic. The aim of the study was to investigate the previous use of screening and preventative measures by women who subsequently were diagnosed by breast cancer. The survey was conducted through a voluntary, anonymous questionnaire survey of 166 randomly selected patients attending an outpatient clinic for patients with breast CA. Main hypotheses of bachelor thesis: 1) More than 50% of the women tested were informed of the recommendation to regularly take part in screening examinations carried out as a part of the breast cancer screening (mammography examination). 2) More than 50% of the females tested regularly (repeatedly) participated in screening examination carried out as a part of the breast cancer screening (mammography examination). 3) More than 50% of the females tested underwent breast examination outside of the breast screening (sonography). 4) More than 50% of the tested females from the group regularly performed self- examination of the breasts (palpation examination). The established hypotheses were not met. However, the result is...
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Application of modern separation techniques in the analysis of biological material
Červinková, Barbora ; Solich, Petr (advisor) ; Mikuš, Peter (referee) ; Štěrbová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Department of Analytical Chemistry Candidate Mgr. Barbora Červinková Supervisor prof. RNDr. Petr Solich, CSc. Title of Doctoral Thesis Application of modern separation techniques in the analysis of biological material The presented dissertation is focused on development, optimization and validation of new extraction procedures and chromatographic methods for determination of vitamins A and E, inflammation biomarkers, DNA and RNA oxidative stress products in biological matrices. In the theoretical part of this work selected analytes and possibility of their determination in biological materials are described. Moreover, modern trends in liquid chromatography and bioanalytical validation guidelines are included. In this part, special attention is focused on sample handling, transport and pre- treatment. The second part is focused on published methods and divided into two sections. The first section comments published developed analytical methods. As first method, determination of lipophilic vitamins in human serum by miniaturized liquid-liquid extraction with subsequent separation on fluorinated stationary phase using UHPLC system with fluorescent detection in 4 min is discussed. Thereafter, determination of creatinine, DNA and...
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Application of modern separation techniques in the analysis of biological material
Červinková, Barbora ; Solich, Petr (advisor) ; Mikuš, Peter (referee) ; Štěrbová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Department of Analytical Chemistry Candidate Mgr. Barbora Červinková Supervisor prof. RNDr. Petr Solich, CSc. Title of Doctoral Thesis Application of modern separation techniques in the analysis of biological material The presented dissertation is focused on development, optimization and validation of new extraction procedures and chromatographic methods for determination of vitamins A and E, inflammation biomarkers, DNA and RNA oxidative stress products in biological matrices. In the theoretical part of this work selected analytes and possibility of their determination in biological materials are described. Moreover, modern trends in liquid chromatography and bioanalytical validation guidelines are included. In this part, special attention is focused on sample handling, transport and pre- treatment. The second part is focused on published methods and divided into two sections. The first section comments published developed analytical methods. As first method, determination of lipophilic vitamins in human serum by miniaturized liquid-liquid extraction with subsequent separation on fluorinated stationary phase using UHPLC system with fluorescent detection in 4 min is discussed. Thereafter, determination of creatinine, DNA and...
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Genetic polymorphism in the HFE gene in the Czech population
Červinková, Barbora ; Beránek, Martin (advisor) ; Šimůnek, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Červinková Supervisor: Doc. PharmDr. Martin Beránek, Ph.D. Title of diploma work: Genetic polymorphism of the HFE gene in Czech population Outline: The main objective of the thesis was to map the incidence of the three mutations (C282Y, H63D, S65C) in the HFE gene in the Czech population. Consequently, the obtained results were compared with the reported occurrence in the world. Methods: The work involved the isolation of DNA from buccal swabs obtained from 167 donors (65 men, 102 women; average age for men was 31, for women 28). Isolated DNA strings were amplified by PCR methods using 5'-CAG ATC CTC ATC TCA CTG-3' and 5'-CTG GAT AAC CTT GGC TGT ACC CCC-3' primers for C282Y mutation, 5'-GCC ACA TCT GGC TTG AAA TT-3' and 5'-ACA TGG TTA AGG CCT GTT GC GCC ACA- 3' primers for H63D and S65C mutations. DNA samples were treated with restriction enzymes Rsa I, Bcl I and Hinf I, for C282Y, H63D and S65C mutations, respectively. Finally, the restriction fragments were separated by gel electrophoresis (2 % agarose gel). Results: C282Y mutation was present in 0 (0 %) and 19 (11,38 %) samples as homozygote or heterozygote, respectively. Moreover, C282Y mutation was present in 3 (1,80 %)...
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