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HPLC method devolopment for betacarotene determination in nutraceuticals
Hlaváčková, Markéta ; Šatínský, Dalibor (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Markéta Hlaváčková Consultant: Doc. RNDr. Dalibor Šatínský, Ph.D. Diploma Thesis Title: Development of HPLC Method for the Determination of Betacarotene in Nutritional Supplements A new HPLC method was optimized and developed for the determination of betacarotene contents in nutritional supplements Betakaroten Farmax, GS Betakaroten FORTE (GreenSwan pharmaceuticals), Walmark Betakaroten, Bioaktivní karoten (Pharma Nord Denmark), Beta karoten Max, BETAVID + LUTEIN (Naturvita), Karovit (Vitabalans Oy), SELZINK PLUS (PRO.MED.CS), Pupalkový olej (Aromatica). The method is based on using Ascentis Express C18 column (30 x 4.6 mm; 2.7 µm) and UV detection at 450 nm. There was used isocratic elution of the mobile phase 100% methanol at a flow-rate of 1.5 ml/min. Column oven temperature was set at 60 řC during the measurement. The retention time of betacarotene under the optimized and validated conditions was 2.185 min. Keywords: betacarotene, HPLC, Betakaroten Farmax, GS Betakaroten FORTE (GreenSwan pharmaceuticals), Walmark Betakaroten, Bioaktivní karoten (Pharma Nord Denmark), Beta karoten Max, BETAVID + LUTEIN (Naturvita), Karovit (Vitabalans Oy), SELZINK PLUS (PRO.MED.CS), Pupalkový olej...
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Study of influence of mobile phase composition on selectivity and retention of the analytes on HILIC stationary phases
Štětková, Kateřina ; Kočová Vlčková, Hana (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Kateřina Štětková Supervisor: RNDr. Hana Vlčková, Ph.D. Title of Diploma thesis: Study of influence of mobile phase composition on selectivity and retention of the analytes on HILIC stationary phases This graduation thesis deals with an influence of concentration of ACN, pH and buffer concentration in mobile phase on retention of selected analytes and on selectivity of Atlantis HILIC Silica column (3 μm, 2,1 mm x 100 mm) and Luna NH2 column (3 μm, 2,0 mm x 100 mm). A set of 35 analytes including neutral, basic and acidic compounds were chosen for this study. Detection was performed by PDA detector at the wave-length of 245 nm. Measurements were carried out using isocratic flow 0.4 ml.min-1 of mobile phase ACN/water (buffer). The column temperature was set up to 30 řC, auto sampler temperature to 4 řC and injection volume to 2 µl. For easier evaluation of results, measured data are presented in charts and tables. These were used to evaluation of columns in terms of selectivity and retention of the individual groups of analytes. The typical HILIC behavior, an increasing retention of analytes with increasing ACN concentration in mobile phase, was observed for Atlantis HILIC Silica column....
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Development of UHPLC method for determination of salicylic acid, methylsalicylate and ethylsalicylate in pharmaceutical formulations
Kosáčková, Nikola ; Matysová, Ludmila (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Nikola Kosáčková Supervisor: doc. PharmDr. Ludmila Matysová, Ph.D. Title of diploma thesis: Development of UHPLC method for determination of salicylic acid, methylsalicylate and ethylsalicylate in pharmaceutical formulations New chromatografic method for determination of salicylic acid, methylsalicylate and ethylsalicylate by using ultra-high performance liquid chromatography was developed. Analysis was carried out using the Kinetex® 1.7 µm XB-C18 100 Å, 50 x 2.1 mm column. The mobile phase was composed of aqueous solution of 1% acetic acid (pH 2.6) and methanol, and the flow rate was 0.4 ml/min. Separation was achieved using gradient elutions. An UV detector at the wavelength of 240 nm was used. Propylparaben was chosen as the internal standard. The total analysis time did not exceed 7 minutes. This method was used for the determination of releasing of salicylic acid and methylsalicylate from matrices consisting of polyester of D,L-lactic acid, glycolic acid and 8% dipentaerythritol (8D) and their dissolution media, which have been prepared at the Department of Pharmaceutical Technology at the FaF UK. Key words: Ultra-High Performance Liquid Chromatography, salicylic acid, methylsalicylate,...
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Development and validation of HPLC methods for determination of preservatives in pharmaceutical formulations
Harmáčková, Barbora ; Matysová, Ludmila (advisor) ; Chocholoušová Havlíková, Lucie (referee)
5 ABSTRACT Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Barbora Harmáčková, DiS. Supervisor: doc. PharmDr. Ludmila Matysová, Ph.D. Title of the diploma thesis: Development and validation of HPLC methods for determination volume of preservatives in pharmaceutical formulations This diploma thesis is focused on the development and validation of methods for determination of preservative content in medicinal products, especially thiomersal and benzalkonium chloride. The development was based on the HPLC methods used for benzalkonium chloride a mixture of methanol and 7.5 mM potassium phosphate in a ratio of 68:32 and for a thiomersal mixture of methanol and phosphate buffer in a ratio of 65: 35: 0.9. The development of the methods for both mentioned substances consisted mainly in optimizing the composition of the mobile phase and the temperature conditions of the analysis. In particular, the length of the analysis and the resolution and symmetry of the peaks of the substances being determined were monitored. In the final version of the HPLC method for benzalkonium chloride an isocratic elution with the mobile phase of MetOH: K3PO4 (pH 3, 7.5 mM) in a ratio of 73: 27 was chosen at 50 ř C. In the final version for thiomersal, an HPLC method with the same...
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HPLC method development for artificial colorants determination in green beer samples
Stachová, Ivana ; Šatínský, Dalibor (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Bc. Ivana Stachová Supervisor: Doc. RNDr. Dalibor Šatínský, Ph. D. Title of diploma Thesis: HPLC method development for artificial colorants determination in green beer samples HPLC method was used and validated for the simultaneous determination of synthetic water- soluble dyes: E 102 - Tartrazine, E 104 - Quinoline Yellow, E 110 - Sunset Yellow, E 131 - Patent blue, E 132 - Indigo carmine, E 142 - Green S, E 133 - Brilliant Blue FCF and E 143 - Fast Green FCF. The method was applied for direct determination of these dyes in samples of green beers Jarní pivo 11ř (Primátor, Náchod), Krasličák 14ř (Ježek, Jihlava), Zelený král Vratislav 12ř (Konrád, Vratislavice), Velikonoční speciál 14ř (Starobrno, Brno), Velikonoční ležák 12ř (Radniční pivovar, Jihlava) Rohozec 11ř (Rohozec, Malý Rohozec), Zelená nefiltrovaná 12ř (Žlebské Chvalovice), Velikonoční zelený ležák 12ř (Purkmistr, Plzeň), Velikonoční kopřivová 12ř (Modrá Hvězda, Dobřany), Valášek 12ř (Valášek, Vsetín), Podkováň 12ř (Podkováň, Dolní Cetno). Analytical Chromolith Performance CN 100 x 4.6 mm column and guard column Chromolith CN 5 x 4.6 mm Merck were used and mobile phase contained 40% (v/v) methanol / 2% (v/v) acetic acid...
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The Use of Nanofibers as a Sorbent for Solid Phase Extraction
Raabová, Hedvika ; Chocholoušová Havlíková, Lucie (advisor) ; Šatínský, Dalibor (referee)
Department: Department of Analytical Chemistry Candidate: Hedvika Raabová Supervisor: PharmDr. Lucie Chocholoušová Havlíková, Ph.D. Title of the diploma thesis: The Use of Nanofibers as a Sorbent for Solid Phase Extraction The thesis is focused on the ability of polyamide 6 nanofibers to efficiently sorb chemical compounds with various physico-chemical properties and on the use of polyamide 6 nanofibers as a modern sorbent for solid-phase extraction (SPE). Analytes from the groups parabens, steroids and flavonoids were chosen as model analytes based on previous studies. Fenoxycarb, permethrin and hydroxypyrene were also tested because polyamid 6 demonstrated good effectivity for lipophilic compounds during the experiment. The nanofibers for this experiment were prepared using Nanospider™ technology at the Technical University in Liberec. There were made three types of polyamide 6 nanofibers with different surface weight. The effect of these various surface properties was also evaluated in the study. The chromatography conditions were optimized for each group of substances individually. The nanofibers were placed into an empty SPE cartridge and the following extraction of analytes was made using a SPE manifold. The obtained eluates were analyzed with HPLC- UV system. Evaluation of the extraction...
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