New photometric assay for chymotrypsin in stool.
In this new photometric assay fecal samples are pretreated with detergent and high concentrations of salts. The subsequent kinetic enzyme determination step involves the chromogenic substrate succinyl-Ala-Ala-Pro-Phe-4-nitroanilide. The sample pretreatment assures a nearly complete solubilization of the formerly particle-bound enzyme, thus permitting determination of the enzyme activity in either the suspension or the supernate after centrifugation. Furthermore this pretreatment enhances the enzyme activity and decreases the Km value. Results by this assay correlate well with those by classical titrimetry and the method is easily adapted to automated systems.
D001322:Autoanalysis; D002021:Buffers; D002122:Calcium Chloride; D002863:Chromogenic Compounds; D002908:Chronic Disease; D002918:Chymotrypsin / Q000032:analysis*; D005243:Feces / Q000201:enzymology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009842:Oligopeptides / Q000378:metabolism; D010195:Pancreatitis / Q000201:enzymology; D000644:Quaternary Ammonium Compounds; D012016:Reference Values; D012965:Sodium Chloride; D012995:Solubility; D013053:Spectrophotometry
D002021:Buffers; D002863:Chromogenic Compounds; D009842:Oligopeptides; D000644:Quaternary Ammonium Compounds; D012965:Sodium Chloride; C043307:succinyl-alanyl-alanyl-prolyl-phenylalanine-4-nitroanilide; D002918:Chymotrypsin; C022806:dodecyltrimethylammonium; D002122:Calcium Chloride