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Microchip‐based capillary electrophoresis for determination of lactate dehydrogenase isoenzymes
Author(s) -
Zhuang GuiSheng,
Liu Jing,
Jia ChunPing,
Jin QingHui,
Zhao JianLong,
Wang HuiMin
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200600506
Subject(s) - capillary electrophoresis , chromatography , chemistry , lactate dehydrogenase , isozyme , electrophoresis , l lactate dehydrogenase , capillary action , biochemistry , enzyme , materials science , composite material
This article describes a novel microchip‐based capillary electrophoresis and on‐column enzymatic reaction analysis protocol for lactate dehydrogenase (LDH) isoenzymes with a home‐made xenon lamp‐induced fluorescence detection system. A microchip integrated with a temperature‐control unit is designed and fabricated for low‐temperature electrophoretic separation of LDH isoenzymes, optimal enzyme reaction temperature control, and product detection. A four‐step operation and temperature control are employed for the determination of LDH activity by on‐chip monitoring of the amount of incubation product of NADH during the fixed incubation period and at a fixed temperature. Experiments on the determination of LDH standard sample and serum LDH isoenzymes from a healthy adult donor are carried out. The results are comparable with those obtained by conventional CE. Shorter analysis times and a more stable and lower background baseline can be achieved. The efficient separation of different LDH forms indicates the potential of microfluidic devices for isoenzyme assay.