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AssigneeBoehringer Mannheim GmbH
InventorsWerner Guthlein, Walter Rittersdorf, Hugo Tiedemann, Peter Vogel, Wolfgang Werner
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The detection and determination of (a) hydrogen peroxide or of a substance which reacts with the formation of hydrogen peroxide or (b) of peroxidase or a peroxidatively active substance, is carried out by contacting an unknown sample with a chromogen and, in case (a), with a peroxidatively active substance or in case (b) with hydrogen peroxide or a substance which reacts with the formation of hydrogen peroxide, and evaluating the coloration produced by the reaction, wherein, as the chromogen, there is used at least one 3,3',5,5'-tetraalkylbenzidine compound of the formula: ##STR1## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4, which can be the same or different, are straight-chained or branched alkyl radicals containing up to 6 and preferably up to 4 carbon atoms.
Method for the determination of (a) hydrogen peroxide or of a substance which reacts with the formation of hydrogen peroxide or of (b) peroxidase or a peroxidatively active substance which comprises contacting an unknown sample with an absorbent carrier sequentially impregnated with first (i) a solution of, in case (a), peroxidase or a peroxidatively active substance dissolved in a water-containing solvent or, in case (b), hydrogen peroxide (or a substance which reacts with the formation of hydrogen peroxide) dissolved in a water-containing solvent and, secondly, (ii) a solution of a chromogen dissolved in an organic liquid containing solvent, and the absorbent carrier being dried after said sequential impregnation, and evaluating the coloration produced by the reaction, wherein, as the chromogen, there is used 3,3', 5,5'-tetraalkylbenzidine compound of the formula ##STR3## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4, which can be the same or different, are straight-chained or branched alkyl radicals containing up to 6 carbon atoms.
Diagnostic composition for the determination of hydrogen peroxide or of a substance which reacts with the formation of hydrogen peroxide which composition comprises an absorbent carrier sequentially impregnated with first, a water-containing solution of peroxidase or a peroxidatively active substance and, secondly, an organic liquid containing solution of a chromogen compound of the formula ##STR4## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4, which can be the same or different, are straight-chained or branched alkyl radicals containing up to 6 carbon atoms,
Diagnostic composition for the determination of peroxidase or of a peroxidatively active substance which composition comprises an absorbent carrier sequentially impregnated with, first, a water-containing solution of hydrogen peroxide or of a substance which reacts with the formation of hydrogen peroxide, and, secondly, an organic liquid containing solution of a chromogen compound of the formula ##STR5## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4, which can be the same or different, are straight-chained or branched alkyl radicals containing up to 6 carbon atoms,
Process for making a diagnostic composition for the determination of (a) hydrogen peroxide or of a substance which reacts with the formation of hydrogen peroxide or of (b) peroxidase or a peroxidatively active substance which comprises sequentially impregnating an absorbent carrier with first, (i) a solution of, in case (a), peroxidase or peroxidatively active substance dissolved in a water-containing solvent, or, in case (b), hydrogen peroxide (or a substance which reacts with the formation of hydrogen peroxide) dissolved in water-containing solvent and, secondly, (ii) a solution of a chromogen dissolved in an organic liquid containing solvent, and then drying the absorbent carrier after said sequential impregnation, wherein said chromogen is of the formula ##STR6## wherein R.sub.1, R.sub.2, R.sub.3 and R.sub.4, which can be the same or different, are straight-chained or branched alkyl radicals containing up to 6 carbon atoms.
Dependencies are inferred from the claim text.