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HPLC in nutraceuticals analysis of chlorogenic acids
Majorová, Michaela ; Šatínský, Dalibor (advisor) ; Matysová, Ludmila (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Michaela Majorová Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: HPLC in nutraceuticals analysis of chlorogenic acids A new UHPLC method was developed and validated for determination of chlorogenic acids and their di-substituted derivates in nutraceuticals and the developed method was used for determination of chlorogenic acids in nutraceuticals Kilostop (Astina Pharm, a.s.), Zelená káva Extra (Medicura natural), Maxivitalis Zelená káva (Simply You Pharmaceuticals), Vieste Zelená káva Premium (Volt Retail), Zelená káva bylinný extrakt (Topvet), Zelená káva (VITO LIFE) a Kyselina chlorogenová (VITO LIFE). The analysis was performed on the Ascentis Express® RP-Amide (100 x 2.1 mm; 2.7 µm) column using gradient elution program with mobile phase consisted of mixture of acetonitrile and 5% aqueous solution of formic acid at flow rate of 0.9 ml/min, the PDA detector wavelenght was set at 325 nm and the column temperature was 30řC. Keywords: UHPLC, chlorogenic acid, green coffee extract, RP-Amide column, nutraceuticals
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Application of liquid chromatography for determination of selected amino-acids and agmatine in biological material
Hazuková, Niké ; Kujovská Krčmová, Lenka (advisor) ; Matysová, Ludmila (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Niké Hazuková Supervisor: Assoc. Prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of diploma thesis: High performance liquid chromatogramy determination of selected amino acids and agamtine in biological material Keywords: agmatine, L-arginine, L-citruline, L-ornitine, UHPLC, chronic wounds This thesis follows up the metod development for the determination of agmatine, L- arginine, L-citrulline, L-ornithine in chronic wounds using ultra-high performance liquid chromatography. The aim of this thesis was to optimize suitable chromatographic conditions for the already mentioned analytes with possibility to use this method in clinical research and practice. Four chromatographic columns were tested. Finally the Meteoric Core C18 BIO the dimensions 100 x 4.6 mm, 16 nm, 2.7 μm (YMC, Germany) was chosen, using these conditions: the mobile phase (MF) was 100% 10 mM AMFO (pH 2.5), the temperature was set at 25 řC, the MF flow rate was 0.25 mL / min, and the injection volume was 2 μL. After optimization of the separation conditions, the method was applied to biological material (exudate from chronic wounds), samples were modified by selected extraction procedures as ultrafiltration and protein precipitation....
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Interaction of phenylpropionic acids with copper
Zemanová, Kamila ; Karlíčková, Jana (advisor) ; Matysová, Ludmila (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Kamila Zemanová Supervisor: PharmDr. Jana Karlíčková, Ph.D. Title of Thesis: Interaction of phenylpropionic acids with copper Copper is an essential trace element which is essential for our body. It has a significant effect on the correct functioning of important organs and it plays an important role in the transfer of electrons at the major enzymatic pathways as a prostetic group. On the other hand, excess or deficiency of copper in the human body can cause many diseases. Phenylpropionic acids are group of substances which can have antioxidant, anti-inflammatory and anticancer effects. They are involved in the scavenging of free radicals and reactive oxygen species. In this diploma thesis, I tested copper chelating activity of six phenylpropionic acids at different pHs by spectrofotometric methods using a hematoxylin and a bathocuproinedisulfonic acid disodium salt as an indicator. 3-(3,4-dihydroxyphenyl)propionic acid showed the highest chelating potential but only using a hematoxylin. KEYWORDS: Copper, Phenylpropionic acids, Antioxidants, Chelating activity, Hematoxylin, Bathocuproin
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Determination of illegal compounds in cosmetics by modern chromatographic methods
Novotná, Aneta ; Khalikova, Maria (advisor) ; Matysová, Ludmila (referee)
Charles University, Faculty of Pharmacy Department of Analytical Chemistry Candidate: Aneta Novotná Supervisor: Dr. Maria Albertovna Khalikova, Ph.D. Consultant: doc. RNDr. Dalibor Šatinský, Ph.D. Title of Graduation Thesis: Determination of illegal substances in cosmetics using modern chromatographic methods This diploma thesis deals with the development of ultra - fast liquid chromatographic method, which will best detect the preservatives used in cosmetics (parabens). The development of the method involves the selection of the best stationary phase and the mobile phase. In the next phase, selecting the appropriate temperature, gradient, flow rate, and before applying real cosmetic samples, validation was required to show that the method is sufficiently accurate and repeatable for this research. This development of the method was performed on the UltiMate ™ 3000 HPLC and UHPLC System, 9 stationary phases from 4 manufacturers (Fortis, Ascentis, Kinetex, Onyx), properties and contents were evaluated. The most suitable stationary phase that separated all 9 substances was Kinetex Biphenyl 100A, 2.6 μm 150 x 4.6 mm. Due to the solubility and properties of the parabens, two mobile phases and one combination thereof (ACN, MEOH) were tested. The acetonitrile-containing mobile phase better separated the...
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Interaction of phenylpropionic acids with copper
Zemanová, Kamila ; Karlíčková, Jana (advisor) ; Matysová, Ludmila (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Kamila Zemanová Supervisor: PharmDr. Jana Karlíčková, Ph.D. Title of Thesis: Interaction of phenylpropionic acids with copper Copper is an essential trace element which is essential for our body. It has a significant effect on the correct functioning of important organs and it plays an important role in the transfer of electrons at the major enzymatic pathways as a prostetic group. On the other hand, excess or deficiency of copper in the human body can cause many diseases. Phenylpropionic acids are group of substances which can have antioxidant, anti-inflammatory and anticancer effects. They are involved in the scavenging of free radicals and reactive oxygen species. In this diploma thesis, I tested copper chelating activity of six phenylpropionic acids at different pHs by spectrofotometric methods using a hematoxylin and a bathocuproinedisulfonic acid disodium salt as an indicator. 3-(3,4-dihydroxyphenyl)propionic acid showed the highest chelating potential but only using a hematoxylin. KEYWORDS: Copper, Phenylpropionic acids, Antioxidants, Chelating activity, Hematoxylin, Bathocuproin
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Method development for determination of 8-hydroxy-2-deoxy guanosine in urine for clinical research
Šestáková, Veronika ; Kujovská Krčmová, Lenka (advisor) ; Matysová, Ludmila (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Veronika Šestáková Supervisor: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of the Diploma Thesis: Method development for determination of 8-hydroxy-2-deoxy guanosine in urine for clinical research This diploma thesis is based on the method development for determination of 8- hydroxy-2-deoxy guanosine, 8-hydroxyguanosine and creatinine using UHPLC. The aim was to develop optimal conditions for clinical research. Experiments were carried out using UHPLC Nexera with mass spectrometer LCMS- 8030, (Shimadzu, Japan). Two stationary phases were tested. The chromatographic separation was achieved using a Meteoric core C18 BIO 4.6 × 50 mm stationary phase with core-shell particles, particle size 2.7 μm (YMC, Germany) secured with a KrudKatcher Ultra 0.5 μm in-line filtr (Phenomenex, Germany). The used mobile phase consisted of water (pH 3 using acetic acid) and methanol (using formic acid 0.2 mM) in the ratio 90:10 (v/v). The temperature was maintained at 25 řC, a flow rate was set at 0.5 mL/min and 4 µl of sample was injected. After optimization of separation conditions, the method was applied to biological material (urine). The samples were prepared using solid-phase extraction. The method...
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Development and validation of UHPLC method for determination of omeprazole in oral suspensions
Reiská, Veronika ; Matysová, Ludmila (advisor) ; Kujovská Krčmová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Anylytical Chemistry Candidate: Veronika Reiská Supervisor: doc. PharmDr. Ludmila Matysová, Ph.D. Title of diploma thesis: Development and validation of UHPLC method for determination of omeprazole in oral suspensions The aim of the diploma thesis was to find out suitable conditions for method which enables to determinate the concentration of omeprazole in oral suspensions. There was used the column Kinetex TM 1.7 µm, C18 100 A, 50 x 2.1 mm, (Phenomenex, USA) for analysis. The mobile phase was 0.025mol/L phosphate buffer and acelonitrile in ratio 74:26. A suitable pH value of 7.6 was chosen. The internal standard method of methylparaben was applied. The whole analysis was detected on the wavelength of 300 nm. After finding of optimal conditions, 6 different samples of oral omeprazole suspensions were measured. The solution of the mobile phase was used for dissolution of samples. The method was validated. Keywords: omeprazole, UHPLC, oral suspensions.
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