PubMed · 5704769
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W F Bowers, R H Haschemeyer. 1968-10-24. A versatile small-volume ultrafiltration cell.. https://doi.org/10.1016/0003-2697(68)90132-2
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A micellar electrokinetic chromatographic technique for determining acid (GZA) and 18 beta-glycyrrhetinic acid (GRA) in human plasma and urine was developed. Sample clean-up and analyte concentration were carried out by solid-phase extraction (with C18 sorbent). The buffer solution used for MEKC contained 20 mM sodium dihydrogenphosphate, 20 mM sodium tetraborate, 20 mM tetrabutylammonium bromide and 50 mM sodium dodecylsulphate, at a pH of 8.7. Good linearities for both GZA and GRA in plasma and urine were obtained. The recoveries of the method were in the range 86.5 to 107%. The detection limits for GZA in urine (0.5 ml) and plasma (1 ml) were 1.6 micrograms ml-1 and 0.8 micrograms ml-1, respectively, while for GRA in urine (0.5 ml) and plasma (1 ml) were 2 micrograms ml-1 and 1 micrograms ml-1, respectively.
The chemical stability of nicotinamide adenine dinucleotide coenzyme (NADH/NAD+) and its derivatives (NADPH/NADP+) was investigated using changes in the UV-visible absorption spectra of these compounds. The spectra of cofactor (reduced form) were monitored at 340 nm wavelength in different buffers, showing a faster degradation in phosphate buffer. This was assigned to the adduct formation between phosphate and NADH (pyridine ring). The three-factor-two-level factorial design study evaluated the contributions: buffers (phosphate and Pipes 0.1 M), pH's (6.8 and 7.8), and temperature (25 and 30 degrees C). The freshly prepared aqueous solutions of coenzyme were analyzed after 40 min under the different conditions (eight assays in triplicate) of the experiment. The main observed effects of the NADH oxidation were, in increasing order, pH, temperature, and buffer without appreciable interactive effects. Therefore, it was verified that the better conditions for cofactor use were alkaline media employing Pipes buffer or its derivatives and low temperatures.
Capillary electrophoresis has been shown to be a very useful analysis technique for secondary metabolites of plants. In the present study a method is described for the qualitative and quantitative determination of ergot alkaloids and their epimers. The extraction from the biological matrix yields recoveries of 50-97%, depending on the individual alkaloid. Using a mixture of 20 mM beta-cyclodextrin (CD), 8 mM gamma-CD, 2 M urea and 0.3% (w/w) poly(vinyl alcohol) to phosphate buffer at pH 2.5 the simultaneous separation of all analytes was achieved. A 37 cm (30 cm) fused-silica capillary, at a voltage of 25 kV and a temperature of 20 degrees C, was used for the analysis. Overall analysis time for the separation was 12 min. The limit of detection of the alkaloids using UV detection at 214 nm can be improved 30-fold to about 9.10(-8) M when laser-induced fluorescence detection is applied.