|
|
Comparison of columns for separation of pharmaceutical substances by method sequential injection chromatografie
Vacková, Jana ; Chocholouš, Petr (advisor) ; Matysová, Ludmila (referee)
Charles University in the city of Prag Pharmaceutical fakulty in the city of Hradec Králové Department of the Analytical Chemistry Consultant: PharmDr. Petr Chocholouš, PhD. Candidate: Jana Vacková Name of the thesis: Comparsion of columns for separation of pharmaceutically significant substances by method of Sequetial Injection Chromatography This diploma thesis deals with the development of the separation method in the system of the Sequential Injection Chromatography (SIC). The used SIC system was based on the system SIChrom™ (FIAlab® , Bellewue, WA, USA) with the eight port stainless steel multi-position valve (VICI® Valco Instruments, TX, USA) and with the pump S17 (Sapphire™ Engeneering, MA, USA). The samples were prepared by dissolving of 25 mg of various phenolic acids (protocatechuic acid, vanillic acid, p-coumaric acid, ferulic acid, o-coumaric acid, sinapinic acid and syringic acid) in 25 ml of methanol. Three fused-core particle columns were used in this thesis : the column Ascentis Express C-18 (30 mm × 4.6 mm, 2.7 µm, Sigma - Aldrich, Supelco Analytical, Bellefonte, PA, USA), the column Ascentis Express RP-Amide (30 mm × 4.6 mm, 2.7 µm, Sigma - Aldrich, Supelco Analytical, Bellefonte, PA, USA) and the column Ascentis Express Phenyl-Hexyl (30 mm × 4.6 mm, 2.7 µm, Sigma - Aldrich,...
|
|
|
Elektro membránová extrakce za nehybných podmínek - Ultrarychlá izolace kyselých léčiv z neupravené lidské plazmy
Prosová, Klára ; Chocholouš, Petr (advisor) ; Sklenářová, Hana (referee)
The presented work describes the development of electro membrane extraction (EME) of acidic drugs from biological samples under completely stagnant conditions. As noticeable from the term, EME across a supported liquid membrane (SLM) is a result of application of an electrical potential difference [1][2]. EME of 3 acidic drugs from untreated human plasma was performed through an organic solvent (decanol) immobilized in the pores of a polymeric hollow fibre (200 µm thickness) as a SLM, and into 10 mM NaOH as the acceptor solution. The driving force for the extractions was electrical potential difference in the range 10-30 V. The influence of the extraction time, voltage and pH of both donor and acceptor solution were investigated and optimum conditions were proposed. Ketoprofen, ibuprofen and naproxen were extracted from plasma samples with recoveries ranging between 2 and 20 % after only 5 to 10 min. No agitation was applied and only low voltage (20 V) was used to drive the target analytes through the SLM. The extraction system provided very efficient sample pre-treatment and pre-concentration prior to separation and detection with capillary electrophoresis-ultra violet light detection (CE-UV) or high performance liquid chromatography (HPLC-UV). Linearity was in the range between 0.5 and 30 µg/ml...
|
|
|
The development of HILIC method for UHPLC-MS/MS determination of pteridins, a comparison of selectivity of various stationary phases.
Jánská, Radka ; Nováková, Lucie (advisor) ; Chocholouš, Petr (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Radka Jánská Supervisor: PharmDr. Lucie Nováková, Ph.D. Title of Doctoral Thesis: The development of HILIC method for UHPLC-MS/MS determination of pteridins, a comparison of selectivity of various stationary phases This graduation thesis was dealing with the development of HILIC method for the identification and quantification of biologically active substances neopterin, biopterin, 7,8- dihydroneopterin and 7,8-dihydrobiopterin by ultra-high performance liquid chromatography coupled to the mass spectrometry detector of triple quadrupole type. Three chromatographic columns (BEH Glycan, BEH Amide and BEH HILIC) were tested. Several mobile phases and their influence on the separation of target analytes were tested. Mobile phase consisted of aqueous component (acetic and formic acid, ammonium formate and acetate and ammonim hydroxide of low concentration) and acetonitrile. On the chromatographic column BEH Glycan following best mobile phases were evaluated: 1mM ammonium acetate pH= 3.8 with acetonitrile in the ratio 30:70 and 1mM ammonium acetate pH= 6.8 with acetonitrile in the ratio 28:72. Two mobile phases composed of 1mM ammonium acetate pH= 4.8 with acetonitrile in the ratio 23:77 and...
|
|
|
Detekce a kvantifikace trinitrotoluenu v reálných vzorcích s použitím metody ELISA
Hanusová, Lucia ; Chocholoušová Havlíková, Lucie (advisor) ; Chocholouš, Petr (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of: Analytical chemistry Candidate: Lucia Hanusová Supervisor in Hradec Králové: PharmDr. Lucie Havlíková Ph.D Supervisor in Bologna: Prof. Steffano Girotti Title of Diploma Thesis: Detection and quantification of trinitritoluene in real samples by using ELISA method. The 2,4,6-trinitrotoluene (TNT) is a widely used explosive. TNT has been used extensively in the manufacture of explosives since the beginning of last century. It could be found as a contaminant in environment, contaminates air, water and soil. It is accumulated in plants and fishes, and could affect human health. A reliable and sensitive method is needed to get the information about pollution in environment, to detect exposion in human organism or to identify the explosive after explosion. A new rapid method of indirect competitive Enzyme-Linked ImmunoSorbent Assay method with chemiluminescent detection (CL-ELISA) for determination of trinitrotoluene in real samples was successfully applied. The immunological analysis using many antibodies was performed after extraction to methanol (MeOH). The antibody labelled with horse-radish peroxidase was added to reaction and after combination with luminescent mixture the light was developed and we were able to...
|
|
|
Vývoj imunochromatografického stanovení pro testování reálných vzorků obsahujících TNT
Kosařová, Lucie ; Chocholouš, Petr (advisor) ; Chocholoušová Havlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Lucie Kosařová Supervisor: PharmDr. Petr Chocholouš, Ph.D. Title of diploma thesis: Development of lateral flow immunoassay for testing real samples containing TNT Obtaining results very fast is requested more often these days. This is particularly true for dangerous and potentially dangerous compounds. One of these compounds is 2,4,6,-trinitrotoluene (TNT), which was tested in this research. TNT has been extensively used as a military explosive and is still one of the most widely used explosives. Its advantages, such as low manufacturing costs, safety of handling and fairly good explosive power can be tempting for abuse by terrorists. In addition, TNT is a well known pollutant because of its toxicity and low biodegradability. It can contaminate surface and ground waters, soils and sediments, thus causing environmental and health problems. According to these facts there is a requirement to detect this nitroaromatic compound in minimal concentrations very fast and outside the laboratory. These demands could be met by the lateral flow test. In this research lateral flow immunoassay (LFIA) based on colloidal gold nanoparticle labels was developed for detection of TNT in real samples. The...
|
|
|
Solid phase extraction by method Lab-On-Valve
Koblasa, Lukáš ; Chocholouš, Petr (advisor) ; Šatínský, Dalibor (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Lukáš Koblasa Supervisor: PharmDr. Petr Chocholouš, Ph.D. Title of diploma thesis: Solid phase extraction by method Lab-On-Valve This thesis deals with the development of method for direct determination of potassium- saving diuretic amiloride in the urine sample by bead injection (BI) using Lab-on-Valve system (LOV). The determination was based on the selective capture of the amiloride on beads with sorbent (CM-Sephadex C-25 ion-exchange), while the possible interfering urine components were flushed into the waste stream of carrier. After preconcentration amiloride was eluted from the beads by eluent solution (0.25 M HNO3 + 0.05 M KCl), with flow rate of 5 µl/s. Detection was carried out a fluorescence spectrometer at a wavelength of 272 nm. Urine was diluted ten times and spiked by amiloride. For one analysis it was necessary to take 300 µl of the sample. Linearity of response was in the range of concentrations from 0.2 to 10.00 µg/ml (r = 0.990). The limit of detection (LOD) corresponds to a concentration of 0.05 mg / ml. The whole analysis lasted 280 seconds.
|
|
|
Solid phase extraction and its miniaturization by method Lab-On-Valve
Krchňáková, Eva ; Chocholouš, Petr (advisor) ; Sklenářová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Bc. Eva Krchňáková Supervisor: PharmDr. Petr Chocholouš, Ph.D. Title of diploma thesis: Solid phase extraction and its miniaturization by method Lab-On- Valve. This thesis deals with the ability to automate solid phase extraction (SPE), in connection with the bead injection (BI). The aim was to test the replacement of microparticles in the bead injection with sorbent from SPE cartridges, determinate the behavior of sorbent in this system and then determine whether it is possible to miniaturize all the steps of solid phase extraction that are necessary to implement the method. In the work was done manual solid phase extraction, where were tested seven sorbents: Lichrolut Merck RP-18, DSC-18 DSC-18Lt, DSC-8, PH-DSC, DSC-CN and Oasis HLB. The experiment resulted in the best possible conditions for carrying out solid phase extraction as follows: activation of the sorbent and elution of analyte 100% acetonitrile, solvation of the sorbent, the dissolution of vitamin D and ballast from the sorbent washed with dilute acetic acid of pH 3. Oasis HLB from Waters Company is the most suitable sorbent for the system, which is water-wettable and should not be dispersed by the system, which was...
|
|
|
Application of the SIA system for aluminium determination
Fialová, Barbora ; Sklenářová, Hana (advisor) ; Chocholouš, Petr (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Barbora Fialová Supervisor: PharmDr. Hana Sklenářová, PhD. Title of Diploma Thesis: Application of the SIA system for aluminium determination This diploma work deals with determination of aluminium ions in samples of surface water in a sequential injection system. For determination of aluminium ions method described in ČSN 83 0520 part 22 B was used. Determination was based on complex formation between aluminium ions and aluminon. The incurred complex is colored and color intensity is directly proportional to aluminium ions concentration in the sample. In this work spectrophotometric detection was used. Primarily, an optimization of reaction conditions in the test tube was performed. The calibration graph was linear in the range 0.02 - 1 mg/l Al with correlation coefficient R = 0.9932. Also detection limit (LOD = 0.0119 mg/l) and quantification limit (LOQ = 0.0390 mg/l) were assumed. Consequently reaction was converted into the SIA system and optimization of reaction conditions in the SIA system was performed. Calibration curve was linear in concentration range from 0.02 to 0.6 mg/l with R = 0.9971. Deformation of calibration curve in the field of higher aluminium concentrations is due...
|
|
| |
|
| |
guest ::
