Determination of trypsin, chymotrypsin and kallikrein in porcine pancreas extracts by capillary zone electrophoresis

摘要

A new method for the direct detection of trypsin, chymotrypsin A and kallikrein from porcine pancreas by capillary zone electrophoresis is proposed. Optimum conditions for quantitative determination were investigated which also allow the unequivocal distinction of trypsin and chymotrypsin according to porcine or bovine source. For each enzyme good linearity and precision were obtained, with a detection limit of 5.0 mug mL(-1). In addition, the formation of the complexes between trypsin and basic pancreatic trypsin inhibitor, as well as that between kallikrein and the same inhibitor, have been investigated. Such complexes can be clearly distinguished either from the unreacted enzyme or the excess inhibitor and their use for analytical purposes has been evidenced. The method is rapid and sensitive and has been applied to determine enzymes in several porcine pancreas extracts. The levels of the active compounds in various kinds of commercial preparations were easily determined with relative standard deviations ranging between 3.3 and 4.9%, and recoveries from 95.8% to 102.4%. [References: 30]