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US 4,294,923 AGranted
Substrates and method for determining enzymes
OwnerRobert E. Smith, Eugene R. Bissellinventors
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Abstract
A method is disclosed for determining the presence of an enzyme in a biological fluid, which includes the steps of contacting the fluid with a synthetic chromogenic substrate, which is an amino acid derivative of 7-amino-4-trifluoromethylcoumarin; incubating the substrate-containing fluid to effect enzymatic hydrolysis; and fluorometrically determining the presence of the free 7-amino-4-trifluoromethylcoumarin chromophore in the hydrolyzate.
Claims14
A method for determining the presence of an enzyme in an enzyme-containing analyte, comprising:
- (a) contacting the analyte with a substrate which can be hydrolyzed by said enzyme to liberate 7-amino-4-trifluoromethylcoumarin, said substrate having the formula ##STR6## wherein R is an amino acid, a peptide, an amino acid which has been reacted with a blocking group, or a peptide, the terminal amino acid of which has been reacted with a blocking group, thereby forming an analyte-substrate mixture;
- (b) incubating the analyte-substrate mixture under enzyme-hydrolyzing conditions to form an enzyme hydrolyzate;
- and
- (c) fluorometrically or spectrophotometrically determining the presence of 7-amino-4-trifluoromethylcoumarin in the enzyme hydrolyzate.
1 more element
Dependencies are inferred from the claim text.