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HPLC Analysis of Vitamines Soluble in Water Using Electrochemical Detector
Jesenský, Ivan ; Kastner, Petr (advisor) ; Mokrý, Milan (referee)
High performacne liquid chromatography is valued analytical method, which is one of the most frequently used in pharmacy. Biotine is at water soluble vitamin, which is an important element of various enzyme, stuff necessary in metabolism. It part take on metabolism of sugar, fat and protein. It occur in several sort of food, especially in roast, peanut butter, egg yolk, carrot, peas, soya, wheat gemmule and oaten flake. This work deals with optimalization chromatographic condition on determination of biotine. Its target is to find optimal mobile phase, near which give out best separation biotine from others element in solution, to find internal standard and form hydrodynamic voltamogramms, individual at water soluble vitamin. For determinatiom was used coulometric detector Coulochem III. Analysis overshoot on column Lichrospher 60, RP - select B (240 x 4,0 mm and.d., 5 em), Merck. Mobile phase was composed of acetic acid : acetonitrile (CH3COOH 0,2410 mol/l; pH 3,0) 85: 15. Flow rate was 1ml/min, in temperature 30žC. Current inserted on working electrode was 900 mV, sensitiveness 200mV.
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HPLC analysis of drugs IV.
Štichová, Lucia ; Sochor, Jaroslav (advisor) ; Mokrý, Milan (referee)
HPLC analysis of drugs IV. Diploma Thesis Lucia Štichová Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control, Heyrovského 1203, Hradec Králové In this thesis, conditions were optimized for high-performance liquid chromatography for quantitative evaluation of ibuprofen. It was established solid phase microextraction conditions for which ibuprofen was extracted from whole blood. The fiber was coated with polydimethylsiloxane - divinylbenzene with a thickness of 60 µm. The fiber was immersed in the sample. The blood sample was adjusted to pH 2.6. Sorption time was set at 30 minutes and the methanol desorption time was 15 minutes. The mobile phase consisted of methanol with water in the ratio 8:2 and pH was adjusted to a value of 3. The flow rate was 1 ml / min and detection was carried out at a wavelength of 222 nm. For quantitative evaluation the external standard was used. The detection limit was 0.005 µg / ml and the quantification limit was 0.017 µg / ml.
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Validation of HPLC Method for Determination of some Vitamins in a Multivitamin Product
Stehlíková, Kristýna ; Mokrý, Milan (advisor) ; Kastner, Petr (referee)
Charles University in Prague Faculty of pharmacy in Hradec Králové Department of pharmaceutical chemistry and drug analysis Candidate: Mgr. Kristýna Stehlíková Consultant: RNDr. Milan Mokrý, CSc. Title of Thesis: Validation of HPLC method for determination of some vitamins in a multivitamin product. A method for simultaneous determination of concentrations of vitamins B1 (thiamine), B3 (nicotinamide), B6 (pyridoxine) and vitamin C (ascorbic acid) in a multivitamin product by means of HPLC was validated. Linearity, precision (carried out as repaetibility), accuracy, specificity and robustness were used as validation parameters. Linearity was carried out with the assistance of method of dilution of standard solutions of vitamins in range 50 - 150% of expected concentrations of particular vitamins. Solution of a multivitamin was prepared six-times for the precision experiment and then relative standard deviation was calculated. The relative standard deviation stayed in the range of 0.9 -1,61%. Accuracy was verified by analysis of model samples prepared by addition of known amount of particular vitamins to placebo. Yield varied between 97-105%. During the specificity experiments it was not found any interefence among the other compounds presented in the multivitamin product. Necessity of keeping given...
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Development of stability indicating method for chosen preparation
Chmurová, Klára ; Kastner, Petr (advisor) ; Mokrý, Milan (referee)
1 Abstrakt Development of stability indicating method for chosen preparation(HPLC determination of benzyl alcohol in vaginal cream) Graduation thesis Klára Chmurová Charles University in Prague, Pharmaceutical Faculty in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control The task was to find out a method of simultaneous determination of benzylkalcohol and imidazol (decomposition product of main substance of cream - clotrimazole). This thesis is intent on determination of benzylalcohol. We were looking for proper modification of the pharmaceutical preparation (vaginal cream), then optimisation of chromatographic conditions and validation of this HPLC method also. The best results were obtained with LiChrosphere RP 18,5 µm column (125 mm x 4,6 mm, SUPELCO, ), methanol : buffer (solution of 250 ml water, 1,05 ml triethylamine and 0,333g natrium hexasulphonate, pH = 7, set up by 85% H3PO4) 30 : 70 (v/v) was used as the mobile phase, flow rate was 1 ml/min. Detection was realised by UV detector with wave length set up on 223 nm. Time of analysis was shorter than 10 min.
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Determination of exogenic compounds in blood by HPLC
Trávníčková, Petra ; Sochor, Jaroslav (advisor) ; Mokrý, Milan (referee)
DETERMINATION OF EXOGENIC COMPOUNDS IN BLOOD BY HPLC Diploma Thesis Petra Trávní ková Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control, Heyrovského 1203, Hradec Králové This thesis is engaged in HPLC analysis of tenoxicam in blood. For sample pretreatment solid phase extraction was used. As the mobil phase the mixture of metanol:phosphoric acid 0,1% (2:1), pH 2,95 was employed. Samples were chromatographed on Separon SGX C18 150 x 3,0 mm I.D. (7µm) column. The UV detection was realized at wavelenght of 280 nm, the fluorimetric detection at excitation wavelenght 350 nm and emission at wavelenght 450 nm. The naproxen was selected as internal standard. The calibration curves were constructed and were verified using three samples. The method has been validated from the point of view of specificity, precision, limit of detection and limit of quantification.
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Evaluation of the chosen active substance in the preparation by UHPLC II.
Husáková, Šárka ; Kastner, Petr (advisor) ; Mokrý, Milan (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Candidate: Šárka Husáková Supervisor: PharmDr. Petr Kastner, Ph.D. Title of thesis: Evaluation of the chosen active substance in the preparation by UHPLC II. Method for the determination of sodium picosulfate monohydrate, its related substances and sodium benzoate using ultra high performance liquid chromatography (UHPLC) was validated. This method has been transmitted from the conditions of previously validated and optimized high performance liquid chromatography (HPLC) method. Separation of the substance was achieved with Kinetex® 2.6 micron C18, 100A, 150 x 3.00 mm, 2.6 microns column using UV detection at a wavelength of 263 nm. The mobile phase consisted of buffer, acetonitrile and propan-2-ol in a ratio of 55: 43: 2 (v/ v/v). The buffer contained sodium dihydrogen phosphate, cetyltrimethylammonium bromide and water. The pH 7.0 of the buffer was achieved by phosphoric acid. The column temperature was 40 ř C, injection volume 3 μl, the flow rate was set at 0.5 ml/min. The method was evaluated as sufficiently selective, precise, exact, linear and sensitive. Furthermore robustness was tested according to the Plackett-Burman design. It was found, that the acetonitrile volume...
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