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The use of HPLC technique in the analysis of phenolic compounds in plant material of fruit trees
Adamcová, Anežka ; Šatínský, Dalibor (advisor) ; Sklenářová, Hana (referee) ; Vávra, Radek (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Anežka Adamcová Supervisor: Prof. RNDr. Dalibor Šatínský, Ph.D. Title of the Dissertation Thesis: The use of HPLC technique in the analysis of phenolic compounds in plant material of fruit trees Although fruits are well studied and known for their high content of antioxidants and positive health benefits, the main aim of the presented dissertation thesis consists in the characterization of biological active compounds in individual parts of fruit tree material (leaves, bark, buds, flowers, and wood chips). The theoretical part is focused on fruit trees and the characterization of phenolic compounds. Their properties and the effects based on current scientific studies are shown. A large part of thesis is dealing with the chromatographic methods for the separation of phenolic substances including the current trends. The extraction process is also included. The practical part deals with the validation and development of screening HPLC-DAD method for the analysis of monitored phenolic compounds (phloridzin, phloretin, chlorogenic acid, quercitrin, rutin, arbutin, 1,5-dicafeoylquinic acid, and 3,5-dicafeoylquinic acid) in individual parts of fruit trees. The main objective of this thesis is to...
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Sequential injection chromatography - advanced and two-dimensional separations
Gužiňák, Matej ; Chocholouš, Petr (advisor) ; Šatínský, Dalibor (referee)
Department: Department of analytical chemistry Candidate: Matej Gužiňák Supervisor: doc. PharmDr. Petr Chocholouš, Ph.D. Title of the diploma thesis: Sequential injection chromatography - advanced and two- dimensional separations The subject of this work was the development of two analytical methods intended for separating substances contained in silymarin, their comparison and preparing a template in the Microsoft Excel program to simplify data evaluation. In the first developed method, called classical sequential injection chromatography (1D- SIC), a long Ascentis® Express F5 chromatography column (10 cm x 4.6 mm, 5 µm) with an opti-guard® C18 pre-column (1 mm) was used during analysis. The mobile phase consisted of a mixture of acetonitrile with phosphoric acid solution with pH 2.0 prepared in a ratio of 27:73. The isocratic mode was chosen for the analysis; the total length was 16.3 minutes with a flow rate of 5 μL/s. In the second developed method, called two-dimensional sequential injection chromatography (2D-SIC), a Kinetex® 5 µm EVO C18 column (30 x 3.0 mm) with an opti- guard® C18 pre-column (1 mm) was chosen for the first dimension. In the second dimension, an Ascentis® Express F5 column (30 mm x 4.6 mm, 2.7 µm) was used. The analysis mode was chosen isocratic with a total length of 35.3...
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Development of UHPLC-DAD method for separation of phenolic substances in plant material and extraction optimization
Turčanová, Anna ; Sklenářová, Hana (advisor) ; Šatínský, Dalibor (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Anna Turčanová Supervisor: doc. PharmDr. Hana Sklenářová, Ph.D. Advisor: Mgr. Marcela Hollá Title of the diploma thesis: Development of UHPLC-DAD method for separation of phenolic substances in plant material and extraction optimization This diploma thesis deals with development of UHPLC-DAD method for separation of selected phenolic substances (arbutin, chlorogenic, neochlorogenic, cryptochlorogenic, isochlorogenic A and caffeic acid, cynarin, catechin, epicatechin, rutin, hyperoside, hirsutrin, reynoutrin, guaiaverin, quercitrin and quercetin). UHPLC analysis of these phenolic substances took place on a Luna® Omega Polar C18 100 (150 x 2.1 mm; 1.6 μm) column with fully porous particles. As a part of the optimization method, a column with core-shell particles (Kinetex® Polar C18) and a column with a hybrid stationary phase (YMC-Triart C18 ExRS) were also tested. The sample injection volume was 2 μl, the flow rate has been set to 0.3 ml/min and temperature of the column has been set to 30 řC. As a mobile phase ultrapure water acidified with acetic acid to pH 2.8 in combination with acetonitrile has been used. The total analysis time was 17.5 min and detection was performed using a DAD detector at...
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HPLC in quality control of food supplements based on curcumine extracts
Stříbrná, Veronika ; Šatínský, Dalibor (advisor) ; Solich, Petr (referee)
Charles University, Faculty of Pharmacy in Hradec Kralove Department of Analytical Chemistry Candidate: Veronika Stříbrná Supervisor: prof. RNDr. Dalibor Satinsky, Ph.D. Title of diploma thesis: HPLC in quality control of food supplements based on curcumin extracts In this diploma thesis, a new HPLC method for the simultaneous determination of active components in food supplements, piperine, curcumin, demethoxycurcumin and bisdemethoxycurcumin was developed and validated. The method was used to determine the level of the relevant analytes in the selected food supplements, namely Curcumin - piperine plus Vieste, Curcumin-piperine complex Natural Medicaments, Turmeric: piperine Setaria, Cur-Cum Astina, Curcumin Liftea, Curcumin Advance a Curcumin turmeric. Finally, the obtained results were compared with the declared extract content and evaluated as a percentage of quality of the food supplement. An Ascentis® Express RP-Amide 150 x 4,6 mm, 2,7 µm particle size analytical column with Ascentis® RP-Amide 5 x 4,6 mm, 2,7 µm particle size precolumn was used for analysis. A gradient elution with analysis time of 12.5 minutes and flow rate of mobile phase 1.0 ml/min was used. As the mobile phase a mixture of acetonitrile 100% and 0.1% phosphoric acid was chosen. The detection was performed with a DAD...
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HPLC in quality control of food supplements containing gingerols and curcumins
Hujová, Eliška ; Šatínský, Dalibor (advisor) ; Chocholouš, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Eliška Hujová Supervisor: prof. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: HPLC in quality control of food supplements containing gingerols and curcumins In this diploma thesis, a new HPLC method for determination of 6-, 8- and 10-gingerol, 6-shogaol, curcumin, demethoxycurcumin, bisdemethoxycurcumin, and piperine was developed and validated. The developed method was used for the analysis of food supplements: Kurkumin - piperin plus (+ zázvor) from VIESTE, Bio tumeric + ginger from VANAVITA, Ginger root extract from BioMedical, Shogaol zázvor from EPIGEMIC®, Ginger root extract from SWANSON®, and Ginger root (Zingiber officinale) from SOLGAR®. Just as a matter of interest, this method was applied for the selected tea blends containing ginger and/ or tumeric: Dilmah Orange & Ginger (flavoured ceylon black tea), Loyd Brusinky & Zázvor, Teekanne Dýchací cesty a krk se zázvorem, Pickwick Nos a krk se zázvorem, Tesco Ginger & Lemon, Teekanne Ginger Curcuma. For the analysis YMC-Triart C18 ExRS (150 x 4.6 mm; 3 µm) analytical column was used. Separation was carried out by gradient elution with mobile phases A: acetonitrile, B: ultrapure water at flow rate 1 ml/min. Detection was...
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Nano-fibers as a modern extraction sorbents for extraction of samples in chromatographic analysis
Raabová, Hedvika ; Šatínský, Dalibor (advisor) ; Ševčík, Juraj (referee) ; Ventura, Karel (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Training workplace Department of Analytical Chemistry Doctoral degree program Pharmaceutical Analysis Candidate Mgr. Hedvika Raabová Supervisor prof. RNDr. Dalibor Šatínský, Ph.D. Advisor assoc. prof. PharmDr. Lucie Chocholoušová Havlíková, Ph. D. Title of Doctoral Thesis Nanofibers as modern extraction sorbents for sample extraction in chromatographic analysis Extraction techniques represent one of the most common approaches to sample handling. The analytes of interest can be isolated easily even from complex matrices, especially by solid-phase extraction (SPE). Therefore, this technique became very popular and routinely used in laboratories. The variety of commercially available sorbents has made this method even more attractive. Nevertheless, the new sorbent materials are still looked for because of the broadening spectrum of analyzed substances and the increasing demands on selectivity and extraction efficiency. Nanomaterials have been increasingly associated with these properties over the last ten years. Therefore, they are receiving more and more attention related to extraction procedures. The dissertation thesis is focused on the application of nanofibrous polymers as innovative sorbents for solid-phase extraction. First, the...
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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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Testing of Chromatographic Qualities of Alternative Stationary Phases
Havlátová, Michaela ; Šatínský, Dalibor (advisor) ; Nováková, Lucie (referee)
1 Abstract Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Michaela Havlátová Consultant: Doc. RNDr. Dalibor Šatínský, PhD. Diploma Thesis Title: Testing of Chromatographic Qualities of Alternative Stationary Phases Chromatographic qualities of HS F5 column were tested at selected groups of substances. For testing there were chosen substances from the following groups: beta-sympatholytics (acebutolol, bopindolol), neuroleptics (thioridazine, perphenazine), local anesthetics (lidocaine, prilocaine), tetracyclins (oxytetracyclin, minocyclin), methylxanthine derivates (caffeine, theophylline) and aniline. During the analysis the retention of the tested substances was investigated as influenced by the character of the mobile phase. Some of the substances showed U-shaped retention. Three series of analysis were carried out at each tested group. In analysis number 1 acetonitrile - water was used as the mobile phase, in analysis number 2 the mobile phase was acetonitrile - phosphoric acid 0.085%, and in analysis number 3 it was acetonitrile - acetate buffer solution pH 7. In each analysis the acetonitrile percentage in the mobile phase was changed. In the group of the substances tested the U-shaped retention was shown by beta-sympatholytic...
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