INFLUENCE OF SEVERAL ANALYTICAL CONDITIONS ON WHOLE BLOOD CHOLINESTERASE ACTIVITY DETERMINATION BY ELLMAN METHOD
Abstract
The main sources of variation that can produce variability on whole blood cholinesterase determinations were studied and evaluated. For this purpose, dog whole blood aliquots were analyzed under different analytical conditions. An increase on cholinesterase activity was observed as reaction temperature was raised. The highest cholinesterase activity was obtained at pH range between 8.0 and 8.5; however, non-enzimatic substrate hydrolysis was also increased when pH exceeded 7.5, so the use of pH 7.5 would be recommended. Although acetylthiocholine iodide concentration of 5x10-3 M and butyrylthiocholine iodide concentration of 10x10-3 M yielded the highest cholinesterase activity, an increase of non-enzymatic substrate hydrolysis was also observed at these substrate concentrations, so the use of both substrates at 1x10-3 M concentration was preferred. 5,5’- dithiobis-2-nitrobenzoic acid as chromophore, and acetylthiocholine iodide and butyrylthiocholine iodide as substrates, were stable for at least three months if frozen. Based on these results, this work in contributing to a better knowledge of the sources of variation affecting whole blood cholinesterase determination, and the standardization of analytical conditions.Downloads
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