Assays for serum cholinesterase activity by capillary electrophoresis and an amperometric flow injection choline biosensor.
A capillary electrophoresis method and a durable choline biosensor were developed for measuring serum cholinesterase (EC 3.1.1.8) activity, a useful clinical index for liver function. The former is based on separation of benzoate and benzoylcholine (the artificial substrate of cholinesterase) in an uncoated fused-silica capillary. The migration time of benzoylcholine and benzoate was 1.3 min and 5.5 min, respectively. By the peak areas of A(233) signals, the linear dynamic ranges for both analytes were 0.01-50.0 mM, and the relative standard deviations of 1.0 mM benzoylcholine and benzoate were less than 4% and 6%, respectively. The FIA-choline sensor was constructed with the working electrode of the flow cell covered with a natural chitinous membrane purified from Taiwanese soldier crab, Mictyris brevidactylus. The biomembrane served as the supporting material for enzyme immobilization (choline oxidase, EC 1.1.3.17), and also prevented protein adsorption on the electrode surface. The calibration curve was linear between 0.05 and 5.0 mM (r=0.999). The relative standard deviations for 1.0 mM choline (n=7) were less than 3%, and the activity of the bioactive membrane lasted for about 2 months. The analytical results of both methods correlated well (r=0.940).
Hsieh
Bo-Chuan
BC
Department of Bio-Industrial Mechatronics Engineering, National Taiwan University, 136 Chou-Shan Road, Taipei City 106, Taiwan, ROC.
Hsiao
Hsien-Yi
HY
Cheng
Tzong-Jih
TJ
Chen
Richie L C
RL
D000818:Animals; D001565:Benzoates / Q000302:isolation & purification / Q000378:metabolism; D001588:Benzoylcholine / Q000302:isolation & purification / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation*; D002686:Chitin / Q000737:chemistry; D002794:Choline / Q000378:metabolism*; D002802:Cholinesterases / Q000097:blood* / Q000378:metabolism; D004563:Electrochemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D017022:Flow Injection Analysis / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D013997:Time Factors
D001565:Benzoates; D008567:Membranes, Artificial; D002686:Chitin; D001588:Benzoylcholine; D002802:Cholinesterases; D002794:Choline