Abstract
Arginine-esterase activities of trypsin, plasmin and kallikreins were assayed by high performance liquid chromatography (HPLC) with BAEE (or TAME) and DAME. 1. In the case of conventional method with BAEE (or TAME), only the hydrolyzed product of BA (or TA) was eluted from the column of "Zipax" SCX (20 cm×2.0 mm i.d.) with 0.1M phosphate buffer (pH 7.0), while BAEE (or TAME) was retained on the column. 10-3-10-2 TAME units of these enzymes were easily measurable by determination of eluted product at several minutes intervals. 2. Using DAME as a fluorescent substrate, separation of DA from DAME was performed on the column of Hitachi gel #3010 (15 cm×2.0 mm i.d.) by elution of 20% (v/v) Tris-HCl buffer (0.05M, pH 8.5) in methanol. In this condition, 4 pmole of DA was determined and 10-5-10-4 TAME units of these enzymes were measurable.
