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Analysis of diosmin, hesperidin and rutin in nutraceuticals by capillary electrophoresis
Vašíčková, Tereza ; Polášek, Miroslav (referee) ; Pospíšilová, Marie (advisor)
Analýza diosminu, hesperidinu a rutinu v potravních doplňcích metodou kapilární elektroforézy 5/82 Abstract A method based on capillary zone electrophoresis (CZE) with UV detection was developed for simultaneous separation and determination of flavonoids rutin, hesperidin and diosmin. The analysis was performed in a fused-silica capillary with effective length 50 cm, i.d. 50 µm, voltage - 30 kV and 25 o C. UV detection was used at 200 nm, 246 nm and 280 nm. The samples were loaded hydrodynamically at a pressure of 50 mbar for 6 s. A single analysis took less than 6 minutes. The separation was optimized by examining a number of experimental conditions, such as concentration of the electrolytic system, pH, addition of organic solvents and cyclodextrins. The optimal background electrolyte consisted of 35 mM natrium tetraborate with 2.5% of methanol (adjusted to pH* 9.0 with boric acid) and 1.5 mM α-CD. The calibration graphs were linear for both rutin (100.58 - 1005.80 µg/ml; r = 0.9991), hesperidin (90.33 - 903.29 µg/ml; r = 0.9785) and diosmin (45.56 - 455.60 µg/ml; r = 0.9972). Propylparaben was chosen as the internal standard. The method was applied to the assay of the flavonoids in a nutraceutical HEMOSTOP® PROBIO, capsules and characterized by RSD 3,25% (rutin), 4,12% (hesperidin) and 3,98% (diosmin), n...
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Analysis of cough-cold preparations by electrophoretic methods.
Kubíková, Drahomíra ; Jáč, Pavel (referee) ; Pospíšilová, Marie (advisor)
In this study, micellar elektrokinetic chromatography method (MEKC) was developed for the determination of paracetamol (PA), caffein (CO), ascorbic acid (AA) and phenylephrine (FE) in a pharmaceutical preparation. Separation was carried out in a fused silica capillary (31,2 cm x 50 m i.d. effective length 21 cm) at 20 kV with UV detection at 190 nm and 265 nm. Optimized background electrolyte (BGE) was 100 mM boric acid containing 100 mM sodium dodecyl sulfate (SDS) as a surfactant and 10 % (v/v) of methanol. pH* 7,8 was found to be optimal (modified by 50 mM sodium tetraborate). Methylparaben (MP) was used as internal standard. This validated method has been successfully applied for the analysis of commercially available pharmaceutical product (Coldrex tbl.).
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Determination of septonex in pharmaceutical preparations by capillary zone electrophoresis with conductivity detection.
Šindelková, Martina ; Polášek, Miroslav (advisor) ; Pospíšilová, Marie (referee)
DETERMINATION OF SEPTONEX IN PHARMACEUTICAL PREPARATIONS USING CAPILLARY ZONE ELECTROPHORESIS WITH CONTACTLESS CONDUCTIVITY DETECTION A new method of capillary zone electrophoresis with contactless conductivity detection for the determination of septonex in pharmaceutical preparations was devised. Optimal conditions for the separation and determination of septonex were: background electrolyte 30mM MES of pH 7.0 (adjusted with 20mM TRIS) containing 12.5mg/ml of (2- hydroxypropyl)-β-cyclodextrin, voltage 25kV, temperature 25řC and sample injection for 15 seconds under the pressure of 50mbar. N,N-dimethylethanolamine (200µg/ml) was used as internal standard. The peak of septonex was satisfactorily separated from the peak of internal standard as well as from the EOF. The analysis was carried out in a fused-silica capillary (internal diameter 50μm, total length 75cm and the length to the detector 45cm). The separation took less than 4 minutes and the overall analysis time involving appropriate rinsing of the capillary was less than 16 minutes. The calibration curve was linear in the range 75µg/ml - 300µg/ml of septonex, correlation coefficient r = 0.9976. The LOD was 9μg/ml and LOQ was 30μg/ml of septonex. Unsuitable repeatability of peak areas of septonex (caused probably by insufficient elimination of...
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HPLC method development for determination of active compounds in Arpalit preparation.
Švestková, Petra ; Šatínský, Dalibor (advisor) ; Pospíšilová, Marie (referee)
A HPLC method was developed for the separation and determination of the substances fenoxycarb and permethrin. The method is based on using HS F5 column (10 x 4 mm, 3 μm particle) and UV detection at 225 nm. The compounds were separated using isocratic elution of the mobile phase acetonitril - water (65:35) at a flow-rate of 1,0 ml/min. There was temperature 70 řC during the measurement. The system enabled successful separation of both compounds in time less than 5 min. The retention time of fenoxycarb was 1,53 min and the retention time of permethrin was 3,68 min. The chromatographic resolution between both compounds was 11,012. The method was applied to analysis of the active substances fenoxycarb and permethrin in veterinary preparations Arpalit® Neo mechanical spray, Arpalit® Neo spray a Arpalit® Neo foam. Developed method was compared with the method available on Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague (HPLC, Chromolith Performance RP-18, 100 x 4,6 mm, gradient elution of mobil phase acetonitril + water/acetonitril (60:40), flow rate 1,0 ml/min, temperature 30 řC, analysis time 11 min). Keywords: fenoxycarb, permethrin, HPLC
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Use of micellar electrokinetic chromatogramy for the assay of degradation products of the pharmaceutical compounds
Vančura, Luboš ; Pospíšilová, Marie (advisor) ; Polášek, Miroslav (referee)
Use of micellar electrokinetic chromatography for the determination of pharmaceutical impurities A new method of micellar electrokinetic chromatography (MEKC) for the separation and selective determination of clotrimazole and its degradation product (2- chlorophenyl)diphenylmethanol in pharmaceutical preparations was developed and validated. The analysis was carried out in a fused-silica capillary (internal diameter 75 µm, total length 60 cm, effective length 50 cm) with UV detection at 210 nm, voltage 30 kV, hydrodynamic sample injection at 34.5 mbar for 7 s and temperature 25 o C. The optimum background electrolyte consisted of 15 mM potassium dihydrogen phosphate, 20 mM sodium dodecyl sulphate, 30 mM sodium cholate and 10 mM β-cyclodextrin (pH = 8.2); sodium diclophenac was used as internal standard. The separation took less than 12 min.
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The analysis of statines by gas chromatography
Pavlovičová, Soňa ; Šatínský, Dalibor (advisor) ; Pospíšilová, Marie (referee)
Gas chromatography analysis of statins Soňa Poláková (roz. Pavlovičová) Abstract A new GC method for determination of statins after derivatizacis using FID detector was developed. During the experiments the following results were achieved. Optimum separation results in the analysis of statins achieved in the following temperature gradient: one minute at 250 ř C, the temperature is further increased at 10 ř C / min to a final temperature of 320 ř C with other isothermal sections. A flame ionization detector was kept at the temperature 320 ř C. The optimum derivatization reagent was chosen BSTFA + TMCS for simvastatin, and N- tert-butyl (dimethylsilyl) trifluoroacetamide for atorvastatin. Addition of anhydrous potassium carbonate was obtained in alkaline environment, which allowed the derivatization. The best temperature for the derivatization of both drugs was obtained at 90 ř C. Time that is necessary to be run derivatization is 60 minutes with simvastatin and atorvastatin for 30 minutes. Quantification was performed in both cases using mefenamic acid as an internal standard, which was also derivatized, in the same derivatization agent. IS derivatization proceeded at 60 ř C for 60 minutes. The greatest yield response was achieved using half the amount of derivatization reagent than the amount of drug. Due...
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Utilisation of electromigration methods in analysis of naturally occuring compounds
Honegr, Jan ; Pospíšilová, Marie (advisor) ; Urbánek, Marek (referee)
UTILISATION OF ELECTROMIGRATION METHODS IN ANALYSIS OF NATURALLY OCCURING COMPOUNDS Jan Honegr ABSTRACT A transient isotachophoresis-capillary zone electrophoresis (tITP-CZE) method has been developed for pre-concentration and determination of nine analytes: caffeic, chlorogenic, o- coumaric, p-coumaric, ferulic and protocatechuic acid, and kaempferol, quercetin and rutin. The effects of several factors such as control of EOF, pH and concentration of the running buffer, addition of organic solvents and their concentrations, addition of cyclodextrins, the duration of injection and the λ max of UV detection were investigated to find the optimum conditions. Under these conditions, the analytes were separated within 15 min. Linearity was evaluated for concentration range 2.5-37.5µg/ml with R = 0.9924-0.9983; the detection limits (S/N 3:1) ranged from 203ng/ml to 5.556µg/ml. The relative standard deviations of the migration times (peak areas) were between 0.79 and 1.01% (1.34 and 2.13%).
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Analysis of ketoprofen, methylparaben and propylparaben by micellar electrokinetic chromatography
Balcar, Josef ; Pospíšilová, Marie (advisor) ; Polášek, Miroslav (referee)
1. ABSTRACT ANALYSIS OF KETOPROFEN, METHYLPARABEN AND PROPYLPARABEN BY MICELLAR ELECTROKINETIC CHROMATOGRAPHY Josef Balcar Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Analytical Chemistry, Heyrovského 1203, Hradec Králové In this study, a new micellar electrokinetic chromatography (MEKC) has been developed for the simultaneous analysis of ketoprofen, methylparaben and propylparaben. Separations were carried out in a fused silica capillary (50 cm × 75 μm i.d.) at 30 kV with UV detection at 200 nm. The optimal background electrolyte was 50 mM tricine buffer (pH* 8.3) containing 30 mM sodium dodecyl sulfate as a surfactant and 15 % (V/V) methanol. Rectilinear calibration ranges were 10.0 - 200 mg/100ml for ketoprofen, 0.2 - 4.0 mg/100ml for methylparaben and 0.1 - 2.0 mg/100ml for propylparaben. The MEKC method was applied to the determination of the analytes in pharmaceutical preparation. The total analysis time was <13 min.
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Determination of pindolol by sequential injection analysis with chemiluminescence detection
Havelková, Jana ; Polášek, Miroslav (advisor) ; Pospíšilová, Marie (referee)
The oxidation of the drug pindolol, [1-(indol-4-yloxy)-3-(isopropylamino)-2-propanol (I)] by permanganate in the medium of aqueous H2SO4 is accompanied by the emission of chemiluminescence (CL) radiation. The CL signal is enhanced by hexametaphosphate. This CL reaction was used for devising automated sequential injection analysis (SIA) assay of I in pharmaceutical preparations. The PC-controlled SIA setup consisted of a Cavro XL 3000 2.5- ml syringe pump, Vici-Valco 10-port selection valve and Spectra-Physics FS970 flow- through fluorescence detector equipped with a lab-made CL detection module. The net CL signal of I increased by a factor of 4.4 (compared to purely aqueous test solution of I injected) if the test solution contained 80% (v/v) of methanol. Optimal order, concentrations and volumes of aspirated zones of reactants were: 43l of 80mM Na hexametaphosphate, 30 l of I in 80% (v/v) methanol, 28 l of 2M H2SO4 and 1 l of 5mM KMnO4. Calibration curve relating the intensity of CL (peak height) to the concentration of I was linear in the range 1 - 10 M I; the limit of detection (S/N = 3) was 0.15 M I. The sample throughput was 90 h-1 . The repeatability of the peak heights was characterized by RSD 1.8% for 30 replicate injections of 8 M I. The SIA-CL method was used for the assay of I in...
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Separation and determination of pharmaceutically important compounds using electrophoretic methods
Hamoudová, Rafífa ; Pospíšilová, Marie (advisor) ; Nobilis, Milan (referee) ; Jokl, Vladimír (referee)
This thesis deals with the use of electrophoretic methods in the analysis of pharmaceutically important compounds. In the theoretical part, the classification of capillary electrophoretic techniques is discussed. Capillary zone electrophoresis (CZE, capillary isotachophoresis (ITP), micellar elektrokinetic chromatography (MEKC), microemulsion elektrokinetic chromatography (MEEKC), isoelectric focusation (IEF), capillary electrochromatography (CEC) and capillary gel electrophoresis (CGE) are described. Further, on-line pre-concentration methods are described. Particular attention is given to the on-line combination of capillary isotachophoresis with capillary zone electrophoresis using column coupling arrangement. This combination enables the improvement of the selectivity of separation and the improvement of the limit of detection of analytes present in complex matrices of biological and environmental origin. Afterwards, the results of the experimental analyses are presented: 1. The first paper deals with the use of CZE in the analysis of ibuprofen and flurbiprofen in pharmaceuticals. Separation was carried out in a fused silica capillary (60 cm x 100 m I.D effective length 45 cm) at 30 kV with UV detection at 232 nm. The optimized background electrolyte was 20mM N-(2- acetamido)-2-aminoethanesulfonic acid...
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