The aim of this study was to develop a high-performance liquid chromatography-tandem mass spectrometry method for the determination of 6-cyanodopamine, 6-nitrodopamine, 6-nitrodopa, 6-nitroadrenaline and 6-bromodopamine in human plasma samples. Strata-X 33 µm solid-phase extraction cartridges were used for the extraction of the catecholamines from human plasma samples. The catecholamines were separated in a 150 × 3 mm Shim-pack GIST C18-AQ column with 3 µm particle size, placed in an oven at 40°C and perfused with 82% mobile phase A (acetonitrile-H2O; 90:10, v/v) + 0.4% acetic acid and 18% mobile phase B (deionized H2O) + 0.2% formic acid at a flow rate of 340 µl/min in isocratic mode. The injected volume was 4 µl and the run lasted 4 min. The method was linear from 0.1 to 20 ng/ml and the lower limit of quantification was 0.1 ng/ml for all analytes. The method was applied to evaluate the plasma levels of catecholamines in plasma of patients with chronic kidney disease and allowed the detection for the first time of circulating levels of the novel catecholamines 6-bromodopamine and 6-cyanodopamine.
We present a new method for estimating the length density (Lv) of convoluted tubular structures exhibiting an isotropic distribution. Although the traditional equation Lv=2Q/A is used, the parameter Q is obtained by considering the collective perimeters of tubular sections. This measurement is converted to a standard model of the structure, assuming that all cross-sections are approximately circular and have an average perimeter similar to that of actual circular cross-sections observed in the same material. The accuracy of this method was tested in eight experiments using hollow macaroni bent into helical shapes. After measuring the length of the macaroni segments, they were boiled and randomly packed into cylindrical volumes along with an aqueous suspension of gelatin and India ink. The solidified blocks were cut into slices 1.0 cm thick and 33.2 cm2 in area (A). The total perimeter of the macaroni cross-sections so revealed was stereologically estimated using a test system of straight parallel lines. Given Lv and the reference volume, the total length of macaroni in each section could be estimated. Additional corrections were made for the changes induced by boiling, and the off-axis position of the thread used to measure length. No statistical difference was observed between the corrected estimated values and the actual lengths. This technique is useful for estimating the length of capillaries, renal tubules, and seminiferous tubules.
Biometry/methods , Image Processing, Computer-Assisted/methods , Models, Biological
