Patent ID: 8741593

Claim:
A method for determining the presence of an enzyme comprising: (a) contacting a biological sample, suspected of containing the enzyme, with a substrate of -formula (V): reacting the compound of formula (V) with said enzyme, if present, to give a product of the formula (X) wherein Di is a fragment having a spectrometric property which property is altered upon cleavage of the C 1 -C 2 bond; and wherein in the compounds of the formulae (V) and (X): (i) R 1 to R 3 , which are identical or different, are H, a substituted or unsubstituted alkyl group, or Di, or R 1 together with Di forms a five-membered aliphatic cycle, or R 2 , together with C 1 , C 2 , and Di, forms a cyclic moiety in said substrate; (ii) X and Y, which are identical or different, are O, S, or —NH; and (iii) P 1 and P 2 , which are identical or different, are a hydrogen atom, an acyl group that is substituted by an aryl, alkyl or peptidyl group; a phosphate group; a phosphate ester group; a phosphonate group; a carbamyl group that is substituted by an aryl, acyl or peptidyl group; a glycosyl group; or a sulfate group, provided that not more than one of the groups P 1 and P 2 is a hydrogen atom; (b) oxidizing the compound of formula (X) with an oxidizing chemical agent, thereby cleaving the bond C 1 -C 2 to obtain a product of formula (D) wherein Di, Y and R 3 are as defined above; and (c) spectrometrically determining the presence of the enzyme by observing the alteration of the spectrometric property of Di.