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In this work we have studied the possibility of determining the rate of phytoplankton photosynthesis in situ using a submersible pump-and-probe fluorometer in water areas differing in their trophic level, as well as in climatic and hydrophysical characteristics. A biophysical model was used to describe the relationship between photosynthesis, underwater irradiance, and the intensity of phytoplankton fluorescence excited by an artificial light source. Fluorescence intensity was used as a measure of light absorption by phytoplankton and for assessing the efficiency of photochemical energy conversion at photosynthetic reaction centers. Parameters of the model that could not be measured experimentally were determined by calibrating fluorescence and irradiance data against the primary production measured in the Baltic Sea with the radioactive carbon method. It was shown that the standard deviation of these parameters in situ did not exceed 20%, and the use of their mean values to estimate the phytoplankton photosynthetic rate showed a good correlation between the calculated and measured data on primary production in the Baltic (r = 0.89), Norwegian (r = 0.77) and South China (r = 0.76) Seas.
Primary screening of possible antileishmanial compounds is currently based either on the use of promastigotes, which are not the clinically relevant stage, or on the time-consuming intracellular amastigote test. With the aim of discovering new antileishmanial derivatives, a semi-automatized method using Leishmania axenic amastigotes and a fluorometric growth indicator (Alamar Blue®) was developed. This assay was evaluated in a comparative study of the susceptibility of promastigotes, axenic amastigotes and intracellular amastigotes of Leishmania mexicana with standard drugs used in antileishmanial therapy (meglumine antimoniate, amphotericin B, ketoconazole and allopurinol) and recently developed ether lipid compounds (edelfosine and miltefosine). The results showed that while inhibitory concentration 50s (IC₅₀s) of axenic and intracellular amastigotes were similar for all of the drugs tested, promastigotes and intracellular amastigotes IC₅₀s differed for three of the six compounds. In conclusion, this axenic amastigote model allowed us to combine the ease of in vitro drug screening on promastigote with the reliability of the IC₅₀ determination on intracellular amastigotes.
Two methods of determining the chlorophyll a concentration in the sea have been formulated on the basis of artificially induced fluorescence measured with the aid of submersible fluorometers. The method of statistical correlation is founded on the empirical relationship between fluorescence and chlorophyll concentration. The theoretical model of fluorescence described in Part 1 of this paper (see Ostrowska et al. 2000, this volume) provides the basis of the other method, the physical method. This describes the dependence of the specific fluorescence of phytoplankton on the chlorophyll concentration, a diversity of photophysiological properties of phytoplankton and the optical characteristics of the fluorometer. In order to assess their practicability, the methods were subjected to empirical verification. This showed that the physical method yielded chlorophyll concentrations of far greater accuracy. The respective error factors of the estimated chlorophyll concentration were x = 2.07 for the correlation method and x = 1.5 for the physical method. This means that the statistical logarithmic error varies from −52 to +107% in the case of the former method but only from −33 to +51% in the case of the latter. Thus, modifying the methodology has much improved the accuracy of chlorophyll determinations.
The objective of this work was to use the fluorometric method to study the antioxidative properties of the preparations obtained from the seed coats of red and brown beans. The preparations were obtained by extraction of the polyphenols with 0.5% HCl solution in methanol, vacuum concentration and lyophilisation. The qualitative and quantitative compositions of the preparations were determined. The antioxidative properties of polyphenolic preparations were determined by measuring the fluorescence as produced by the formation of hydroxide radicals from hydrogen peroxide and sodium benzoate in the presence of copper ions and dithioerythritol (DTET) in phosphate buffer, pH 7.2. The fluorescence of hydroxide derivatives was measured spectrofluorometrically. The studies showed that the preparations had the capabilities to scavenge the hydroxide radicals by 51% in the case of brown pigments and by 47% in the case of red pigments.
The range of variability of the fluorescence properties of marine phytoplankton in different trophic types of seas and at different depths in the sea is analysed theoretically. An attempt is also made to interpret artificially induced in situ fluorescence measured with submersible fluorometers. To do this, earlier optical models of light absorption by phytoplankton (see Woźniak et al. 2000, this volume) and actual empirical data were applied. A straightforward theoretical model of artificially photoinduced phytoplankton fluorescence accounting for the complex influence of different photophysiological characteristics of phytoplankton and the optical characteristics of the instrument has been worked out. A physical method of determining chlorophyll a concentrations in seawater from fluorescence measured in situ with contact fluorometers can be based on this model.
The paper contains a preliminary analysis of the links between the portion fc of functional PS2 reaction centres in the photosynthetic apparatus of marine phytoplankton and environmental factors. The analysis is based inter alia on fluorometric measurements of fc (see Kolber & Falkowski 1993) in water sampled from different depths and trophic types of sea. From the statistical generalisations was derived an analytical form of the relationship between fc, and the optical depth and trophic type of sea (the trophicity index was taken to be the surface concentration of chlorophyll a). According to this relationship, fc rises as the trophicity of the sea does so. Moreover, there is a certain optimal optical depth for each type of water at which the number of functional PS2 reaction centres reaches a maximum. Above or below this depth the value of fc falls. At the present stage of investigations it seems reasonable to assume that this drop in the number of functional PS2 reaction centres close to the surface is due to the destructive effect of excessive irradiance. On the other hand, their reduced number at greater depths, below the fc maximum, can be attributed to the deficit of light and the consequent destruction of reaction centres.
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Ocena wybranych metod oznaczania kwasu mlekowego

59%
Przeprowadzono ocenę i porównanie następujących metod oznaczania kwasu mlekowego: miareczkowej, kolorymetrycznej, fluorymetrycznej, enzymatycznej oraz HPLC. Opracowano metodę ilościowego oznaczania kwasu mlekowego w oparciu o technikę HPLC oraz porównano wyniki uzyskane tą metodą z wynikami uzyskanymi przy pomocy innych metod. Dokonano oceny dokładności tych metod w oparciu o odzysk, oraz ocenę precyzji na podstawie współczynnika zmienności.
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