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Photoreactivable 5-l3-(p-iodoazidosalicylamide)allylJ-UDP-GlcNAc (5-IASA-UDP- -GlcNAc) was synthesized by a four-step procedure and used for photoaffinity labeling of UDP-GlcNAc-dependent enzymes. Upon iodination with 12SI, the compound was successfully applied to probe a purified UDP-GlcNAc pyrophos­phorylase from pig liver. The enzyme was photoinactivated by the probe in the concentration-dependent manner, and was protected by UDP-GlcNAc and, to a lesser extent, by UTP and UDP-GlcCOOH.
A simple method for determination of 8 anabolie hormone residues in the muscles and urine of slaughter animals was elaborated. Forty cattle muscle samples and 30 urine samples fortified with diethyl stilbestrol, dienstrol, hexastrol, zeranol, trenbolone, trenbolone acetate, 19-nortesterone, and medroxyprogesterone acetale at concentrations of 1 to 20 ng/g (ppb) were examined. The hormones were extracted with methanol and the methanol phase was degreased with n-hexane and extracted with diethyl ether. The ether layer was washed with carbonate buffer and water evaporated to dryness. The residues were dissolved in acetate buffer and cleaned up by extraction into the solid phase on C18 columns. In urine, the hormones were extracted with diethyl ether followed by the procedure corresponding to that for muscles. After evaporating and dissolving in acetone the samples were analysed by two-dimentional HPTLC methods. The chromatograms were developed with 5% sulphuric acid in ethanol. The detection limit of the hormones examined on the HPTLC plate was 5-20 ng, and in the tissue 1 ppb for trenbolone and 2 ppb for the remaining compounds.
We used thin-layer chromatography of fecal bile acids in an attempt to separate feces of culpeo fox Pseudalopex culpaeus (Molina, 1782) from those of chilla fox P. griseus (Gray, 1837). We tested the method with pure bile acids and with feces from known fox species on two different plate types. Results differed according to the plate type used. The technique failed to distinguish scats from the two fox species. We found variability in the spot pattern of bile acids within species, and within individuals, likely associated to the diet. The location of the spots {Rf values) also varied with the concentration of the sample, and the color and location changed with different plate types. We warn that the thin-layer chromatography of fecal bile acids is still unreliable for distinguishing feces between sympatric carnivores. Hence, we propose to rely on alternative techniques.
A gas chromatographic procedure using methylbicyclophosphorothionate as an internal standard, for the determination of ethylbicyclophosphate in the brain, blood, and liver of small laboratory animals, was elaborated. The compound was extracted from the tissue with acetonitrile, and re-extracted from 2.5% sodium sulfate solution with chloroform. After evaporation of the solvent, the residue was dissolved in acetone. Chromatographic partition was carried out using a glass column (4 m x 0.4 cm) packed with 3% OV-17 on W/H.P. chromosorb at 250°C. Recovery of ethylbicyclophosphate ranged from 65% to 95%. Linear relationship of standard curve within the concentration range from 0.4 to 4.0 µg of ethylbicyclophosphate, was found. Precision for the assay of 2.0 µg samples, was 7.36%.
Clerodendrum phlomidis Linn. f. of the family Lamiaceae is an important and well known medicinal plant in Ayurveda and Siddha system of medicines. To ensure identity, quality of the plant material and considering the wide therapeutic application of L-DOPA, lupeol and β-sitosterol, the present study was planned to quantify these marker constituents by TLC method. The amount of L-DOPA, lupeol and β-sitosterol quantified from the leaves of C. phlomidis were 0.06806, 0.01733 and 0.06324 % w/w, respectively. This TLC procedure may be used effectively for identity, quality evaluation as well as quantitative determination for this plant or its derived products.
Z odtłuszczonych nasion soi wyekstrahowano związki fenolowe 80% (v/v) roztworem metanolu. Uzyskany ekstrakt rozdzielono na sześć frakcji (I- VI) stosując chromatografią kolumnową na żelu Sephadex LH-20 z metanolem jako fazą ruchomą. W uzyskanych frakcjach oznaczono zawartość fenoli ogółem i zarejestrowano widma UV. Frakcje scharakteryzowano również za pomocą chromatografii cienkowarstwowej na żelu krzemionkowym. Aktywność przeciwutleniającą mierzono w układzie modelowym z ß-karo- tenem i kwasem linolenowym.
Celem pracy było przeprowadzenie oznaczeń obecności sulfonamidów w dostępnych w handlu naturalnych miodach pszczelich z zastosowaniem chromatografii cienkowarstwowej (TLC) jako metody przesiewowej do wykrywania tych zanieczyszczeń.
Lipid was extracted from the lipid containing muscle of Liner silver grunter (Pomadasys hasta) by solvent extraction and then purified by standard method. The physico-chemical properties of lipid sample were determined and compared with those of other standard fats and oils. Fatty acid composition of the lipid sample was investigated by Thin Layer Chromatographic (TLC) examination. Gas Liquid Chromatographic (GLC) examination was performed by the methyl esters mixture prepared from the lipid sample. The de-oiled muscle of the fish was studied for the determination of ash content, fiber content, moisture content and protein content. The lipid containing selected muscle was analyzed quantitatively for the determination of percentages of protein, cholesterol and minerals (N, P, K, Ca, and Na). The lipid containing muscle was also analyzed for the determination of the amount of eleven metals by using Atomic Absorption Spectrophotometric method. The lipid sample was evaluated by microbial activity (bacterial activity against nine bacteria and fungal activity against seven fungi).
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