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HPLC separation of phenolic substances in plant material
Majerčíková, Jana ; Sklenářová, Hana (advisor) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Jana Majerčíková Supervisor: doc. PharmDr. Hana Sklenářová, Ph.D. Title of the diploma thesis: HPLC separation of phenolic substances in plant material This diploma thesis is focused on development and optimization HPLC-DAD method for determination of 17 phenolic compounds (gallic acid, protocatechuic acid, 4-aminobenzoic acid, chlorogenic acid, catechin, caffeic acid, epicatechin, vanillin, p-coumaric acid, ferulic acid, salicylic acid, rutin, o-coumaric acid, quercitrin, myricetin, quercetin and kaempferol) in apricot variety Betynka. In the process of optimization several types of stationary phase and gradients of mobile phase were tested. For the analysis of real samples guard column Security GuardTM Polar C18 (4 x 2.0 mm, 3 μm) and column Luna Omega Polar (150 mm x 2.1 x 3 μm) were selected. As mobile phase the mixture of ultrapure water acidified with the acetic acid to pH 2.8 and acetonitrile was used. For the analysis the gradient elution with initial isocratic step was used, total analysis time was 17 minutes. The flow rate was 0.6 ml/min and the injection volume 1 μl. The detection was performed by DAD spectrophotometric detector at wavelengths of 254, 260, 280 and 320 nm. Validate...
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Optimization of neonicotinoids extraction using nanofibers in sequential injection analysis systém
Zouharová, Monika ; Horstkotte Šrámková, Ivana (advisor) ; Šatínský, Dalibor (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Monika Zouharová Supervisor: PharmDr. Ivana Horstkotte Šrámková, Ph.D. Title: Optimization of neonicotinoids extraction using nanofibers in sequential injection analysis system In this diploma thesis, we tested the use of various nanofibers as sorbents for solid phase extraction of neonicotinoids connected with sequential injection analysis system. Seven types of nanofibers were tested, specifically polyamide (PA6), polycaprolactone (PCL), polyimide (PID) and polydopamine coated polyamide (PA6)coated, polycaprolactone (PCL)coated, polyimide (PID)coated and a combination of polyimide and polycaprolactone (PID/PCL)coated. These nanofibers were produced by electrospinning. The studied nanofibers were placed in the 3D printed holder, which was connected into sequential injection analysis system. UV spectrophotometry was used as a detection method. The substances to analyse were neonicotinoid insectisides. The retention of these analytes was examinated with all seven types of nanofibers. Retention of the analytes on the tested nanofibers depending on their chemical structures and physical-chemical properties was studied in these measurements. The most experiments were performed with polydopamine coated...
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Modification of nanofibres for extraction in liquid chromatography
Valachovič, Adam ; Šatínský, Dalibor (advisor) ; Chocholouš, Petr (referee)
Department: Department of Analytical Chemistry Candidate: Adam Valachovič Supervisor: doc. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: Modification of nanofibers for extraction in liquid chromatography The diploma thesis demonstrates the influence of modifications of polymer nanofibers on their extraction properties for extraction in liquid chromatography. Nanofibers represent a potential sorbent for extraction methods due to their large surface area, which leads to increased capacity of extraction. Another advantage of nanofibers is the possibility to modify their surface resulting in various nanofibers with different extraction properties. In this thesis, the modifications were performed by coating polypropylene nanofibers with a polydopamine layer. The polymerization solution was pumped through a column, in which the nanofibers were placed, using a peristaltic pump connected to the column by a system of tubes. An on-line SPE HPLC method was developed to determine the extraction properties of the modified nanofibers. Chlorogenic acid was chosen as the model analyte, because its extraction is affected by the polydopamine layer. The key parameters of the coating process were optimized, including the weight of the nanofibers, the concentration of dopamine in the polymerization...
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Analysis of nutritionally important substances in fruit tree waste products by HPLC
Šírová, Karolína ; Šatínský, Dalibor (advisor) ; Sklenářová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Karolína Šírová Supervisor: doc. RNDr. Dalibor Šatínský, Ph.D. Consultant: Mgr. Anežka Adamcová Title of Diploma Thesis: Analysis of nutritionally important substances in fruit tree waste products by HPLC This diploma thesis deals with the content of phenolic compounds in various parts of tree of the genus Pyrus. The theoretical part is focused on characterization, biosynthesis, chemical and biological properties of major phenolic compounds in pear-trees and the possibilities of their determination with the use of the latest scientific studies. The experimental part contains an extensive analysis of arbutin, chlorogenic acid, 1,5- dicaffeoylquinic acid, 3,5- dicaffeoylquinic acid and rutin in extracts of leaves, bark, buds, fruits, flowers and wood chip. Developed and validated HPLC-DAD method was used and the total antioxidant activity was determined by FIA method with electrochemical detection. An ASCENTIS Express RP-Amide 150×4.6 mm, 2.7 μm analytical column and gradient elution consisting of organic part of acetonitrile (ACN) and water part with 0,85% phosphoric acid (pH = 2.2) were used for analysis. The detection was performed by DAD detector at wavelengths 220 nm, 327 nm and 354 nm. The...
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Application of nanofibrous sorbents for on-line extraction in liquid chromatography
Háková, Martina ; Šatínský, Dalibor (advisor) ; Česla, Petr (referee) ; Urban, Jiří (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Martina Háková Supervisor: doc. RNDr. Dalibor Šatínský, Ph.D. Consultant: doc. PharmDr. Lucie Chocholoušová Havlíková, Ph.D. Title of Doctoral Thesis: Application of nanofibrous sorbents for on-line extraction in liquid chromatography A compilation of ten publications is presented in this dissertation thesis. It is focused on the application of nano and microfibrous polymers as a new extracting material in analytical chemistry. This work was based on close cooperation with the Institute for Nanomaterials, Advanced Technologies and Innovation and the Department of Nonwovens and Nanofibrous Materials at the Technical University of Liberec, where the author took part in designing and preparation of these materials. The first part deals with model studies, based on using polyamide, polystyrene and polycaprolactone nanofibers as sorbents for solid phase extraction in classical off-line mode or in on-line column switching system. It solves practical aspects such as orientation and the formation of the fibers in the extraction system, optimization of fabrication factors affecting applicability of the nanomaterials, suitable solvent systems, stability testing and reusability of the microcolumns. The...
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Optimization of separation conditions for HPCCC of paclitaxel
Havrlantová, Šárka ; Kučera, Radim (advisor) ; Šatínský, Dalibor (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Šárka Havrlantová Supervisor: doc. PharmDr. Radim Kučera, Ph.D. Name of diploma thesis: Optimization of separation conditions for HPCCC of paclitaxel The aim of this work is a feasibility study of natural substances isolation (in my case it is paclitaxel and its intermediate 10-deacetylbaccatin) using modern technique called High performance countercurrent chromatography (HPCCC) available in Teva in Opava. For this basic research it was necessary to study the literature and test various solvent systems. Chosen solvent systems were then tested together with natural substance in HPCCC, results were processed and discussed. Countercurrent chromatography is considered a modern separation technique based on two immiscible liquids. One of them is called a stationary phase and is held inside the column by centrifugal force, the second liquid is called a mobile phase and is pumped through the column. Then the injected liquid sample is also pumped through the column and depending on distribution coefficients the sample components are either held in the stationary phase or pass through the column and detector into fraction collector. Advantages of this method are e.g. total...
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Modern chromatographic techniques in food contamination analysis
Lhotská, Ivona ; Šatínský, Dalibor (advisor) ; Karlíček, Rolf (referee) ; Malíř, František (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate Mgr. Ivona Lhotská Supervisor doc. RNDr. Dalibor Šatínský, Ph.D. Title of Doctoral Thesis Modern chromatographic techniques in food contamination analysis A compilation of seven publications dealing with chromatographic methods for determination of food contamination (particularly mycotoxins and artificial food colorants) is presented in dissertation thesis. All methods were fully validated, complying with the requirements and applied for real samples analysis. The first part describes the methods for mycotoxins determination based on on-line solid phase extraction (SPE) utilizing column-switching system. Reversed phase columns packed with superficially porous particles were chosen for both extraction and separation step during optimization of on-line SPE-HPLC method for ochratoxin A and citrinin determination in beer. In following work, determination of patulin in juices and zearalenone in beer, more selective molecularly imprinted polymers (MIP) were used for on-line extraction. Due to challenging optimization of on-line MIP SPE, the methods were compared to off-line MIP extraction and on-line extraction using conventional C18 phase to confirm the selectivity. The employed commercial MIP...
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Application of HPLC technique in analysis of food supplements based on plant extracts
Fibigr, Jakub ; Šatínský, Dalibor (advisor) ; Cahlíková, Lucie (referee) ; Horna, Aleš (referee)
ABSTRACT Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Jakub Fibigr Supervisor: doc. RNDr. Dalibor Šatínský, Ph.D. Title of the Dissertation Thesis: Application of HPLC technique in analysis of food supplements based on plant extracts The disseration thesis deals with complex issues of food supplements based on plant extracts in terms of the content of bioactive substances, purity assessment, and legislative efficacy and safety requirements. The quality of food supplements available on the market in the Czech Republic was tested in six studies. Two HPLC and four UHPLC methods have been developed and validated to determine the content of bioactive substances in food supplements containing plant extracts and other excipients. In each work, such analytical conditions were found allowing a rapid and accurate determination of the content of the given bioactive substances in food supplements, particulary in the problematic separations of some isomeric substances. Additionally, sample preparation methods for each group of food supplement samples have been developed and optimized to avoid the matrix interferences during the separation and detection of analyzed bioactive substances. A conventional approach to the analytical determination of the...
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Determination of beta-carotene content in sour cherries by HPLC
Svobodová, Markéta ; Šatínský, Dalibor (advisor) ; Sklenářová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Markéta Svobodová Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of the rigorous thesis: Determination of beta-carotene content in sour cherries by HPLC In this rigorous thesis, we determined the presence of carotenoids (trans betacarotene, cis beta-carotene and lutein) in 18 cultivars of sour cherries. Cultivars: Rannaja, Meteor, Krasnyj flag, Pandy 45, Muskatnaja pražskaja, Erdi jubileum, Griot Moskovskij, Valkova, Mesabich, Gorsetin, Inga mladi, Naumburger osheimer, Pandy 6039, Körös, Morellen fever, Wolyňská, Heimann's rubín, and Višňa IV/155. The optimized method was taken from the diploma thesis of Štěpán Růžička - HPLC method development for carotenoids determination in fruits (2017). In the chromatographic method, we used a Supelco Analytical RP-Amide column (100 x 4.6 mm, 5 μm) and carotenoids were detected at a wavelength of 450 nm. Measurements were carried out in the isocratic elution mode with a mobile phase of acetonitrile: hexane: dichloromethane (97.5 : 1.25 : 1.25) at a flow rate of 1.8 ml / min at 30 ř C. The analyzed samples were extracted by ultrasound into chloroform. After centrifugation, they were filtered and injected to the HPLC column. The sample volume was 5...
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Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques
Javorská, Lenka ; Solich, Petr (advisor) ; Pohanka, Miroslav (referee) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department: Department of Analytical Chemistry Candidate: Mgr. Lenka Javorská Supervisor: prof. RNDr. Petr Solich, CSc. Consultant: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of dissertation thesis: Development of bioanalytical methods for the determination of diagnostic markers and drugs using chromatographic techniques The presented dissertation is focused on the development and validation of chromatographic method for the determination of glycopeptide antibiotic vancomycin, and L-arginine and its pathway related substances in biofluids. Furthermore, the potential of neopterin, kynurenine and tryptophan as biomarkers of immune response to surgical procedure and prediction of postoperative complications is investigated in connection with oncological diseases. In the theoretical part trends in instrumentation, stationary phases and extraction techniques are described. The issues of separation and retention of polar compounds and biological matrix pretreatment before analysis are also discussed there. Next part deals with selected analytes and their application to the clinical purpose for which the chromatographic methods are developed. It also provides an overview of published methods for their determination using liquid...
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