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Analytical evaluation of drugs by liquid chromatography VI
Rosecký, Jan ; Sochor, Jaroslav (advisor) ; Mokrý, Milan (referee)
Analytical evaluation of drugs by liquid chromatography VI Degree paper Jan Rosecký Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control, Heyrovského 1203, Hradec Králové The aim of this degree paper was to optimize conditions for the determination of 6-aminocaproic acid, hydrolytic degradation product of accelerant of transdermal penetration transkarbam 12, using high-performance liquid chromatography (HPLC) and subsequently to validate this method. 3,5-dinitrobenzoyl chloride was used before HPLC analysis for derivatization of 6-aminocaproic acid. Analysis of derivatized acid was performed on Separon SGX C-18 analytical column (150 x 3mm I.D, 5µm). The gradient elution was used for the separation. The mobile phase was consisted of acetonitrile and sodium acetate buffer pH 4,5 and the flow rate was 1 ml/min. 20 µl sample volumes were injected into the chromatographic system. Wavelength used for the detection was 230 nm. The method of internal standard was used for quantitative evaluation of derivatized 6-aminocaproic acid and 7-aminocaprylic acid was chosen as the most suitable internal standard. After the development of the method, these validation parametres were evaluated: precision, accuracy, linearity, selectivity,...
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HPLC analysis of drugs III
Blažková, Petra ; Sochor, Jaroslav (advisor) ; Mokrý, Milan (referee)
ANALYTICAL EVALUATION OF ACTIVE SUBSTANCE BY LIQUID CHROMATOGRAPHY II. Diploma thesis Petra Blažková Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control, Heyrovského 1203, Hradec Králové The subject of this diploma thesis is a determination of active substances by liquid chromatography. In particular it deals with an evaluation of phenobarbital in biological matrix. Solid phase microextraction (SPME) and high performance liquid chromatography (HPLC) with UV detector (218 nm) and reversed phase C18 comlumn were used. The mobile phase consisted of methanol and phosphate buffer in a ratio of 50:50. The flow rate was set to 0,4 ml/min. 20 µl of sample solution was injected on the chromatographic column. Phenobarbital was dissolved in a mixture of methanol and buffer in a ratio of 80:20. For the SPME extraction from water solution was used carbowax/TPR fibre. The analyte was desorbed into methanol. The extraction time was 20 minutes. Phenobarbital extraction from rabbit plasma was performed in the same way, but PDMS/DVB fibre instead of carbowax/TPR fibre was used. Phenobarbital in rabbit plasma was quantified by means of calibration curve.
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Analytical determination of active compounds by liquid chromatography III.
Doskočilová, Marie ; Sochor, Jaroslav (advisor) ; Mokrý, Milan (referee)
ANALYTICAL DETERMINATION OF ACTIVE COMPOUNDS BY LIQUID CHROMATOGRAPHY III. Diploma Thesis Marie Doskočilová Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control, Heyrovského 1203, Hradec Králové In this thesis, the development of suitable conditions for analytical sample oxazepam in rabbit blood by high-performance liquid chromatography. For treatment of biological material used solid phase microextraction. Analysis carried out on a C18 column. Mobile phase consisted of methanol and water mixture in the ratio 80:20. UV detector was used in the wavelength of 230 nm. Quantitative evaluation was carried out by internal standard, which was diazepam. Oxazepam was isolated from biological material using SPME. For extraction was used the fiber PDMS / DVB, the desorption medium was methanol. Sorption and desorption time lasted 13 minutes. Extraction efficiency was 5, 9%. For quantitative evaluation of calibration curve was prepared, which were subsequently verified by model samples.
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HPLC analysis of drugs and drug candidates in biological material
Luňáková, Veronika ; Kovaříková, Petra (advisor) ; Mokrý, Milan (referee)
High-performance liquid chromatography (HPLC) is currently one of the most progressive separation methods, permitting, simultaneously, both quantitative and qualitative analysis. An important area for the utilisation of HPLC is the identification of drugs and their metabolites in biological materials. This study was focused on the development of chromatographic conditions appropriate for HPLC analysis of two new potential drugs derived from aroyl-hydrazone, HNAF-INH and 4,6DAR-INH, in biological material and its subsequent application in the assessment of the stability of these substances in plasma. Analysed chelators were prepared by the modification of the structure of salicylaldehyde isonicotinoyl hydrazone (SIH) with the purpose of improving the stability of the substance in plasma. As a suitable stationary phase the HPLC column LiChroCART® HPLC - cartridge LiChrospher®100 RP - 18e (15μm) with the pre-column LiChroCART® 4-4 Purospher® RP - 18 (5μm) was chosen. The mobile phase in this case represented by a HNAF-INH compound: phosphate buffer (0,01 mol/l aqueous solution of NaH2PO4 · 2 H2O with 2 mmol/l EDTA, pH 3) : methanol, 42:58 (v/v). The temperature in the column was 25řC, the flow rate 1,0 ml/min and UV detection was performed at 288 nm; SIH was used as an internal standard. The mobile...
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Validation of HPLC evaluation of amlodipine and atorvastatin in combinated dosage forms
Kašková, Jana ; Mokrý, Milan (advisor) ; Pilařová, Pavla (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Analysis Candidate: Mgr. Jana Kašková Consultant: RNDr. Milan Mokrý, CSc. Title of Thesis: Validation of HPLC evaluation of amlodipine and atorvastatin in combinated dosage forms The topic of this thesis is the validation of chromatographic method. Validated method is the simultaneous HPLC analysis of amlodipine besilate and atorvastatin in combinated dosage form. Chromatografic column Nucleosil 125x4 mm 120-5 C18, Macherey-Nagel was used as stationary phase. Mobile phase was formed by mixture of acetonitrile and dibasic sodium phosphate dodecahydrate solution (0.025 mol / l), acidified with phosphoric acid at pH 4.4 in the ratio 55 : 45. A flow rate was 1.0 ml/min and the temperature was 25 ř C. There was used the internal standard propylparaben. The detection was performed at 210 nm using an UV-VIS detector. The method was validated for linearity, precision, accuracy, robustness and selectivity. Linearity was confirmed by analyzing samples of standards in the range 50-150 % of their expected concentration. To verify the accuracy there were prepared six samples of the same composition and then relative standard deviation was calculated. RSD ranged from 0.04 to 1.38 %. Accuracy was...
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Analytical evaluation of drugs using chromatographic methods VI.
Boudová, Kristýna ; Mokrý, Milan (advisor) ; Kastner, Petr (referee)
Analytical evaluation of drugs using chromatographic methods VI. Diploma thesis Kristýna Boudová Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of pharmaceutical chemistry and drug analysis. There was worked out the method for simultanious determination of vitamins B1 (thiamine), B3 (nicotinamide), B6 (pyridoxine) a vitamin C (ascorbic acid) using high performance liquid chromatoghaphy in multivitamin tablets in this work. This method was used for determination of these vitamins in effervescent multivitamine substance used as food supplement. Acquired results were compared with value declared by the producer. Best results were reached by using chromatographic column Lichrosphere 100 RP C18, 250 mm x 4 mm I.D. (5 µm) Merck, Germany with mobile phase methanol : water (pH adjusted to 2,8 by acetic acid 10%) 0,005 mol/l sodium hexanesulfonic acid + 0,1% triethylamine) in proportion 20:80 (v/v). There was used spectrofotometric detector with wavelenght 270 nm for detection. Flow rate was 1 ml/min, time of analysis didn't exceed 13 minutes. Substances were eluated in order: ascorbic acid, nicotinamide, pyridoxine and thiamine. The method will be validated.
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Development and validation of an HPLC method for determination of 3-aminopropanol and stability study of pharmaceutical preparation
Dittrichová, Kateřina ; Mokrý, Milan (advisor) ; Kastner, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Analysis Student: Mgr. Kateřina Dittrichová Supervisor: RNDr. Milan Mokrý, CSc. Consultant: Ing. Lubomír Galla Title of rigorous thesis: Development and validation of an HPLC method for determination of 3-aminopropanol and stability study of pharmaceutical preparation A new method for the determination of 3-aminopropanol by high- performance liquid chromatography coupled with fluorescence detector was designed and developed. The method was based on derivatization of 3- aminopropanol with flurescamine in solution of sodium tetraborate decahydrate and subsequent formation of fluorescent derivative. Reversed-phase column Discovery C18 (5 µm, 150 mm × 4.6 mm I.D.) was employed. The fluorescence detector used two wavelengths: EM 483 nm and EX 390 nm. The suitable determination was carried out with an isocratic elution of mobile phase composed of acetonitrile and ammonium acetate buffer in the ratio 26:74 (v/v) in the 1.00 ml min-1 as the flow rate, and at the isocratic temperature mode of 30řC. Injection volume was 10 µl. The system suitability test (SST) was performed on three parameters: the retention time, method efficiency, and peak symmetry. The method was then fully validated. The...
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Analytical evaluation of the pharmaceutical formulations using chromatographic methods I
Hladík, Michal ; Mokrý, Milan (advisor) ; Kastner, Petr (referee)
Analytical evaluation of the drugs using chromatographic methods I. (Determination of the conditions for simultaneous evaluation of clotrimazole and benzylalcohol) Thesis Michal Hladík Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Analysis Optimum chromatographic conditions for simultaneous HPLC determination of clotrimazole and benzylacohol in a pharmaceutical formulations (2 % cream) were elaborated. The best results was achieved using chromatographic column Separon SGX CN, 7 μm, 150 x 4 mm I. D. (Tessek Prague, Czech Republic). The mobile phase consisted of mixture of acetonitrile and aqueous solution of K2HPO4 0.05 mol/l (60 : 40), pH 6.7 (adjusted with 10 % H3PO4). The flow rate was 0.8 ml/min. The UV detector set at 268 nm was used. Method was validated for linearity and robustness. Complete validation will be an aim of the next rigorous work.
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