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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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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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Bioanalytical method development for the investigation of the role of pyridoxal isonicotinoyl hydrazone in isonazid intoxication
Effmertová, Marcela ; Kovaříková, Petra (advisor) ; Sochor, Jaroslav (referee)
High-performance liquid chromatography is one of the most frequently used separation methods for an analysis of drugs in biological material. It is suitable both for quantitative and qualitative drug analysis. Isoniazid is used for the treatment and a prevention of tuberculosis. Acute isoniazid intoxication is manifested by tonic-clonic convulsions. Vitamine B6 (pyridoxine) is used in clinical practice as an antidotum. Although this type of toxicity is considered to be associated with the deficiency of vitamine B6 (or its active form, pyridoxal-5-phosphate - PLP), description of mechanisms and processes, which are responsible for this effect, is still missing. The aim of this study was to develop suitable chromatographic conditions for monitoring intoxication by isoniazid and detection of potential hydrazone metabolites. These conditions should allow a separation of seven analytes (isoniazid, acetylisoniazid, pyridoxal, pyridoxol, pyridoxal-5-phosphate, and the o-108 - internal standard). Furthermore, the aim was to study possibilities of isolating all those substances from rabbit plasma. The best chromatographic separation was achieved using the HPLC column LiChroCART 250 × 4 nm I.D., packed with Lichrospher 100 RP-18 (5 µm). The mobile phase was composed of a buffer - component A (NaH2PO4 0.01 mol/l, with...
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HPLC EVALUATION OG FUMAGILLIN IN THE MEDICINE
Strýčková, Kristýna ; Mokrý, Milan (advisor) ; Sochor, Jaroslav (referee)
HPLC EVALUATION OG FUMAGILLIN IN THE MEDICINE Rigorous thesis Mgr. Kristýna Strýčková Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control In this rigorous thesis is described analytic evaluation of fumagillin by using HPCL and stated the amount of fumagillin in the medicine. The measurment was implemented on chromatographic colony in size 3 x 150 mm I.D. with the Seladon SGX NH2 filling, 7μm. There was used the mobile phase composed of methanol: amonium acteate (aqueous solution 0,005 mol/l, v/v )in the ratio of 60:40, at the flow rate of 1ml/min and pressure of 2,5-2,6 MPa. The detection was performed at 335 nm using the UV detektor. The samples were spraeyd in cubage 20 μ/l. The international standard method was drawn up for determination of fumagillin in medicine. The most suitable sample from the selected conditions was nimesulid. Concerning validation parameters were verified linearity, accuracy, robustness, selection and detector limit. Further there was examined the stability in conditions of room temperature and the temperature od 2-8˚C. It was determined 0,0238 g of fumagillin in 1 g of medicine that is stated as 95,2% of neccessary amount.
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