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Determination of Heparin by Sequential Injectin Analysis with Spectrophotometric Detection
Lišková, Růžena ; Hraníček, Jakub (advisor) ; Nesměrák, Karel (referee)
In this work the determination of heparin using Sequential Injection Analysis (SIA) technique with spectrophotometric detection was studied. The principle of this method was measuring the decrease of absorbance of phenothiazine dyes in its maximum wavelength caused by its interaction with heparin. The decrease is proportional to concentration of heparin in sample. The aim of this work was to find optimum conditions for determination of heparin by the SIA with spectrophotometric detection and to try to apply this measurement to real sample under these conditions. Three phenothiazine dyes were used: Methylene Blue, Azure A and Azure B and for each of them an optimum concentration was found. Their values were for methylene blue 4.0 ∙ 10-5 mol/dm3 and 5.0 ∙ 10-5 mol/dm3 for Azure A and Azure B. 150 µl was chosen as an optimum amount of heparin and reagent aspirated. Optimum reaction time was 0 s and flow rate during measurement 1.5 ml/min. Working concentration range was found between 0.23 and 15 mg/dm3 for methylene blue, 1.2 and 13 mg/dm3 for Azure A and 0.67 and 12 mg/dm3 for Azure B. Limit of detection varied between 0.09 - 0.35 mg/dm3 and limit of quantification between 0.30 and 1.2 mg/dm3 for different dyes. Relative standard deviation was determined as a measure of repeatability. Its values were...
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Kinetic determination of nitrites with spectrophotometric detection
Vašíčková, Pavla ; Nesměrák, Karel (advisor) ; Rychlovský, Petr (referee)
Title: Kinetic determination of nitrites with spectrophotometric detection Annotation: The proposal and optimization of the kinetic determination of nitrites in water samples is the aim of this bachelor thesis. The method is based on the ability of nitrites to catalyze the oxidation of toluidine blue by bromate in acidic media. The course of the reaction is monitored by spectrophotometrically, using a decrease of the absorbance band at 653 nm in the constant period of time from the beginning of the reaction. The dual-chanel manifolds FIA apparatus was proposed and optimized. The calibration dependency in the range of 0,05-1,00 mg ml-1 was measured. The instrumental limit of detection of 0,038 mg ml-1 and limit of quantification of 0,125 mg ml-1 were calculated. The method was applied on real sample of river water. Key words: nitrites, flow-injection analysis, kinetic catalytic methods
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Analysis of Historical Pharmaceutical Preparations Containing Alkaloids, Sulphonamide, Derivatives of Barbituric Acid, and Derivative of Pyrazolone
Kudláček, Karel ; Nesměrák, Karel (advisor) ; Bosáková, Zuzana (referee)
Pharmaceutical preparations of quinine (injection solutions), sulfanilamide, aminophenazone, barbital (tablets), caffeine, phenobarbital (dragee), and theophylline (suppositories) about seventy years old were analyzed using RP-HPLC. Samples were quantified by HPLC-UV and UV-spectrophotometry. Products of degradation were identified using HPLC-UV and HPLC-MS. Conditions of separation were optimized. The samples of quinine injection solutions consist of 92% or 87% of declared quinine content. Quinotoxine has been identified as the product of quinine degradation. The quantification of theophylline in suppositories and caffeine in dragee did not show any degradation after more than 67 years from their manufacturing. Decrase of potent amount (decrase about 8-22 %) were found in drugs containing sulfanilamide, barbital, phenobarbital and aminophenazone. Products of degradation of these pharmaceuticals were not found.
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