National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
HPLC determination of gallic acid as a possible product of enzymatic reaction of shikimic acid, NADP+ and SDH.
Smolejová, Jana ; Červený, Václav (advisor) ; Nesměrák, Karel (referee)
This diploma thesis deals with the development of an HPLC method for the determination of selected compounds participating in enzymatic reaction leading to the formation of gallic acid. The analysed reaction mixture contains the following reagents: shikimic acid, NADP+ and shikimatedehydrogenase (SDH) extracted from parsley; the presumed product of the reaction is gallic acid. Two chromatographic methods for the determination of the above mentioned compounds were developed using C18 HPLC column and porous graphitic carbon Hypercarb column. Molecular absorption spectrometric detection in the UV range was used in all measurements. Separation on the C18 column was found particularly suitable for analysing the composition of the end products of the reaction. Because of the NADP+ and shikimic acid peak overlap it is necessary to observe absorbance at 212 and 260 nm. Shikimic acid and NADP+ can be quantified due to the fact that shikimic acid does not absorb at 260 nm while NADP+ absorb radiation at both wavelengths. Separation via Hypercarb column was found particularly suitable for analysing the process of the reaction; additional products or intermediates can be seen in chromatograms compared to the C18 method. Determination with Hypercarb column is characterized by higher sensitivity and lower limit...
Interference study by arsenic determination using UV-photochemical generation of its volatile species with AAS detection
Smolejová, Jana ; Červený, Václav (advisor) ; Nesměrák, Karel (referee)
This thesis deals with the interference study of the UV-photochemical generation of arsenic volatile species with AAS detection and it searches for a suitable reaction modifier, which would improve sensitivity of the As determination. Interferences were classified into 3 groups according to their influence on the As UV-photochemical volatile species generation. Ni2+ , Cu2+ , Cl− and merkaptoethanol belong among the negative interferents. HNO3, Fe3+ , ethanol, SO4 2− , TiO2 and L-cysteine belong among the low-level interferents. Co2+ , acetonitrile, triethanolamine, selenite and Bi3+ belong to a group, which increases the absorbance of arsenic and could, therefore, be called positive interferents. Concentration of 10 mg l−1 of Bi3+ significantly increases the level of absorbance of arsenic. This effect was utilized during measurement of calibration dependence and Bi3+ ions were used as the reaction modifier. The figures of merit of this method with reaction modifier were obtained. Limit of detection was 18 µg l−1 , limit of quantification was 60 µg l−1 , sensitivity was 1.144·10−3 l µg−1 , repeatability was 4.5 % (relative standard deviation) a linear range was 60 - 500 µg l−1 . We achieved eleven times better absorbance by using this reaction modifier compared to UV-photochemical volatile species...

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