Premium
Establishment and development of a CZE–UV method for rapid measurement of aprotinin potency
Author(s) -
He Shujuan,
Zhang Jing,
Dong Yue,
Duan Xiaoyun,
Yang Fatang,
Luo Tian,
Wang Zhen,
Dong Yuming
Publication year - 2020
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201900387
Subject(s) - aprotinin , potency , chromatography , trypsin , substrate (aquarium) , repeatability , chemistry , capillary electrophoresis , phosphate buffered saline , enzyme , biochemistry , in vitro , medicine , oceanography , surgery , geology
Abstract A new method for the measurement of aprotinin potency by CZE–UV detector was established for the first time. The on‐line mixing of substrate, trypsin and aprotinin using at‐inlet technology was realized by the established method. Enzymatic reaction, separation, and detection of substrate and product can be performed simultaneously online. The aprotinin potency can be measured within 4 min. The response surface methodology was used to optimize the incubation conditions of trypsin and substrate, and the optimized conditions were obtained under 17.39 mM phosphate buffer at pH 7.6, 1.40 min of incubation time. The repeatability of proposed method was evaluated in three different systems of capillary zone electrophoresis: (i) only substrate; (ii) trypsin and substrate; (iii) aprotinin, trypsin and substrate, and the RSDs of migration times and peak areas of substrate were less than 2.7 and 3.1%, respectively. The RSDs of migration times and peak areas of product were less than 2.1 and 3.0%, respectively. A formula was also developed to calculate the aprotinin potency in this method. In a word, the established CZE–UV method was convenient, fast, and environmentally friendly for the measurement of aprotinin potency.