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Optimalization and validation of HPLC method for the determination of diclofenac sodium and its degradation product in tablets
Birková, Agáta ; Matysová, Ludmila (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Agáta Birková Supervisor: PharmDr. Ludmila Matysová, Ph.D. Title of Diploma Thesis: Optimization and validation of HPLC method for the determination of sodium diclofenac and its degradation product in tablets This diploma thesis deals with the optimisation and the validation of the HPLC method determination of sodium diclofenac (DF) and its degradation product 1-(2,6-dichlorphenyl)-indolin-2-on (DPI) in tablets. DF is a non-steroidal anti-inflammatory drug with analgesic, antipyretic and anti-inflammatory effects. The degradation product DPI has been produced in formulations after a long-term storage, especially after exposure to light or heat. The method development was based on a method created by the Pharmaceutical Faculty of Charles University for the determination of DF, its degradation product and preservatives in topical emulgel and the method was optimised for determination of DF and DPI in tablets. The monolithic column Chromolith® Performance RP-18e (100×3 mm, Merck) and flurbiprofen as an internal standard were chosen. The mobile phase was prepared of methanol and aqueous solution of phosphoric acid (pH 2,5) in the ratio of 65:35 and the flow rate of 1 ml/min. An UV detector...
Optimalization of sample preparation step for UHPLC-MS/MS analysis of atorvastatin, rosuvastatin and their metabolites
Pilařová, Veronika ; Nováková, Lucie (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Veronika Pilařová Supervisor: Doc. PharmDr. Lucie Nováková Ph.D. Title of GraduationThesis: Optimalization of sample preparation step for UHPLC-MS/MS analysis of atorvastatin, rosuvastatin and their metabolites The purpose of this graduation thesis was the optimalization of UHPLC-MS/MS method for the determination of concentrations of atorvastatin, rosuvastatin and their metabolites in biological material, and then development and optimalization of a MEPS (microextraction by packed sorbent) method for sample preparation of biological material and validation of this method. Acquity BEH C18 column (50 x 2.1 mm, 1.7 µm, Waters) was used for the separation of the analytes. Electrospray ionization was performed in both negative and positive ion mode. Triple quadrupole mass analyser was used for detection. Precursor ions and fragment ions were chosen for each statin. Collision energy and cone voltage were optimized for all analytes individually. Quantification of analytes was performed using the SRM (selected reaction monitoring). Protonated molecule [M+H]+ , which was measured in the positive ion mode, was chosen as a precursor ion for rosuvastatin lactone. Ion [M-H]- , which was measured in...
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...
Nová HPLC metoda pro stanovení nafazolinu v očních přípravcích
Dulavová, Martina ; Chocholoušová Havlíková, Lucie (advisor) ; Matysová, Ludmila (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Martina Dulavová Supervisor: PharmDr. Lucie Havlíková, Ph.D., Ass.Prof. Hannelore Kopelent Title of Diploma Thesis: A New Selective and Stability Indicating HPLC Assay for the Determination of Naphazoline in Preparations for Ocular Use Naphazoline is a 2-imidazolidine derivated drug and alpha-adrenergic agonist with vasoconstrictive and decongestive properties. Naphazoline is indicated for the therapy of rhinitis, sinusitis or allergic conjunctivitis. Naphazoline is used in liquid formulations for ophthalmic and nasal application. Naphazoline is marketed in a number of commercially available products; for example, it is contained in Coldan® Augentropfen. Due to economic and therapeutic reasons, hospital pharmacies produce miscellaneous in-house preparations. For our study, two different preparations manufactured in the sterile production of a hospital pharmacy containing naphazoline are investigated. The first formulation is based on two commercially available products and it is a mixture of Coldan® Augentropfen and Okuzell® Augentropfen at a ratio of 1:9. The second formulation is Bor-Naphazolin Augentropfen and it is completely prepared in a hospital pharmacy. In addition to...
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...
Development and validation of method for determination of clotrimazole in cream using HPLC
Poláčková, Jitka ; Matysová, Ludmila (advisor) ; Chocholoušová Havlíková, Lucie (referee)
The topic of my work was to optimalize and to validate the method for determination of capacity of the Clotrimazol, degradeted product (2-chlorophenyl) diphenylmethanol and conservations (methylparaben and propylparaben) in the preparation Clotrimazol ointment. My effort for this work was to develop the method that could half for total separation the particular defined substance in this sample, in the acceptable time, to find inside standard, to optimalize the conditions of separation and the method verify. The optimum chromatographic conditions were found on column Zorbax SB - Phenyl, 3,5 μm, 4,6 x 75 mm in the flow of mobile phase 0,5 ml per minute and wave - length 210 nm. Composition of the mobile phase was acetonitrile - water (55:45), pH water component was modified by acid phosphoric 5 % on 3,2. If these conditions are used, this method is validating. The testing parameters were accuracy, correctness, linearity, selectivity, robustness, detection and quantitative limit and test if the chromatographic system is acceptable.
Stanovení obsahu residuí kofeinu v povrchových vodách metodou HPLC s UV detekcí
Glosová, Ivana ; Solich, Petr (advisor) ; Chocholoušová Havlíková, Lucie (referee)
The occurence of caffeine residues was investigated in superrficial waters. The aim of this study was to obtain a first overview about contamination of waters from Mondego River and from source fontains near Coimbra city ( Portugal) with caffeine. The samples were preconcentrated by SPE ( Solid Phase Extraction) with Oasis polymeric column procedure and analysed by Liquid Chromatography (LC) with UV detektor ( 280nm). Mobile phase consist of 190 ml acetic acid glacial 100% and 810 ml of LC water (adjusted to pH 3.0 with acetate sodium) A total of twelve samples were collected in different site of Mondego River and from fountains near Coimbra city during three month's periods. A total of twelve samples of caffeine were analysed under the conditions described and none of them showed to be contaminated with caffeine.
Development and validation of HPLC method for the determination of ketoprofen in suppositories
Zahálka, Lukáš ; Matysová, Ludmila (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Lukáš Zahálka Supervisor: PharmDr. Ludmila Matysová, Ph.D. Title of Diploma Thesis: Development and validation of HPLC method for the determination of ketoprofen in suppositories The purposes of diploma thesis were development and validation of HPLC method for the determination of ketoprofen in Ketonal® 100 mg suppositories. Ketoprofen belongs to NSAID and it is used for symptomatic therapy of inflammatory, degenerative and metabolic rheumatic diseases and for palliative therapy of some urgent or chronic painful syndromes. During development of determination of ketoprofen in suppositories were used two methods, which were developed at the Faculty of Pharmacy in Hradec Králové. Method for determination of diclofenac in suppositories was default for sample preparation and method of HPLC determination of ketoprofen, methylparaben and propylparaben in gel was basic for conditions of HPLC analysis. Development of method consisted in modification of origin sample preparation method; chromatographic conditions have not been changed. During the experimental work was found to determine what influence has the various steps of sample preparation and extraction temperature. The time of extraction...
Monitoring of Retention "U-profiles" of Selected Substances on Pentafluorophenyl Stationary Phase
Havlátová, Michaela ; Šatínský, Dalibor (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Analytical Chemistry Department Candidate: Mgr. Michaela Havlátová Consultant: Doc. RNDr. Dalibor Šatínský, PhD. Rigorous Thesis Title: Retention "U" profiles testing of selected substances on pentafluorophenyl stationary phase Retention "U" profiles were examined on pentafluorophenyl stationary phase at selected substances. For testing there were chosen substances from the group of tetracycline antibiotics (oxytetracycline, minocycline), neuroleptics (thioridazine, perphenazine), local anesthetics (lidocaine, prilocaine), methylxanthine derivates (caffeine, theophylline), ß-sympatholytics (acebutolol, bopindolol) and aniline. During the analysis the retention profiles of the tested substances were observed as influenced by the character of methanol mobile phase. Three analysis series were carried out at all of the tested substances. In analysis series number 1 the mobile phase methanol - water was used. For analysis series number 2 it was the mobile phase methanol - phosphoric acid 0,085% and for analysis series number 3 it was the mobile phase methanol - acetate buffer solution pH 7. In each analysis the methanol percentage in the mobile phase was changed. In the group of the tested substances the U-shaped retention profile was...
Study of influence of mobile phase composition on selectivity and retention of the analytes on HILIC stationary phases
Štětková, Kateřina ; 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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