Process for the determination of the ph value in the...

G - Physics – 01 – N

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150/13, 260/468.

G01N 33/84 (2006.01) C07C 121/75 (1985.01) C07C 125/075 (1985.01)

Patent

CA 1194888

ABSTRACT The determination of the pH value in the interior of a cell is accomplished through measurement of the fluores- ence emission of a fluorescent substance which is absorbed by the cell through incubation in a solution containing the fluorescent substance. The fluorescent substance is a substance whose emission spectrum has a pH-dependent frequency at maximum emission, and is employed in the form of a derivative adapted to penetrate the cell membrane and to be split in the cell by enzymes present in the cell such that the fluorescent substance is liberated inside the cell and fluoresces as a function of the pH value therein. The frequency at which the fluorescence emission maximum occurs is determined through measurement of the fluorescence of the liberated fluorescent substance. A fluorescent substance particularly suitable for this purpose is 2,3-dicyano-1,4- hydroquinone. Suitable membrane-penetrating derivatives are 1,4-diacetoxy-2,3-dicyanobenzene, 1,4-dibutyryloxy-2,3- dicyanobenzene and 1,4-di-(tert-butyloxycarbonyl-1- alanyloxy)-2,3-dicyanobenzene.

394985

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