1993 Volume 9 Issue 1 Pages 15-17
An automated enzyme analysis was designed for trypsin and Chymotrypsin. The technique consisted of a hydrophobic chromatography of the enzyme and coupled postcolumn substrate reaction. In a desalted aqueous medium, trypsin and chymotrypsin were adsorbed on a phenyl modified adsorbent gel by hydrophobic interaction, and eluted by the concentration gradient of an electrolyte. After the substrate for the enzyme was reacted in a postcolumn reaction coil, the product was spectrophotometrically detected at 410nm. The separatory determination of trypsin and chymotrypsin was carried out by using the suitable substrate solution. Each calibration plot was a good straight line at 5-100μg of enzyme. The relative standard deviation of 50μg trypsin and the same quantity of chymotrypsin was 2% and 3.5% (n=5, within-run) respectively. The method was applied to the determination of trypsin and chymotrypsin activity of rat pancreatic juice.
