A non-oxidative electrochemical approach to online measurements of dopamine release through laccase-catalyzed oxidation and intramolecular cyclization of dopamine.
A new electrochemical approach to selective online measurements of dopamine (DA) release in the cerebral microdialysate is demonstrated with a non-oxidative mechanism based on the distinct reaction properties of DA and the excellent biocatalytic activity of laccase. To make the successful transition of the distinct sequential reaction properties of DA from a conceptual determination protocol to a practical online analytical system, laccase enzyme is immobilized onto magnetite nanoparticles and the nanoparticles are confined into a fused-silica capillary through an external magnetic field to fabricate a magnetic microreactor. The microreactor is placed in the upstream of the thin-layer electrochemical flow cell to efficiently catalyze the oxidation of DA into its quinonoid form and thereby initialize the sequential reactions including deprotonation, intramolecular cyclization, disproportionation and/or oxidation to finally give 5,6-dihydroxyindoline quinone. The electrochemical reduction of the produced 5,6-dihydroxyindoline quinone at bare glassy carbon electrode is used as the readout for the DA measurement. The laccase-immobilized microreactor is also found to catalyze the oxidation of ascorbic acid (AA) and 3,4-dihydroxyphenylacetic acid (DOPAC) into electroinactive species and, as such, to eliminate the great interference from both species. Moreover, the successful transition of the mechanism for DA detection from the conventional oxidative electrochemical approach to the non-oxidative one substantially enables the measurements virtually interference-free from physiological levels of uric acid, 5-hydroxytryptamine, norepinephrine, and epinephrine. The current response is linear with DA concentration within a concentration range from 1 to 20 microM with a sensitivity of 3.97 nA/microM. The detection limit, based on a signal-to-noise ratio of 3, is calculated to be 0.3 microM. The high selectivity and the good linearity as well as the high stability of the online method make it very potential for continuous monitoring of cerebral DA release in physiological and pathological processes.
Lin
Yuqing
Y
Beijing National Laboratory for Molecular Sciences, Laboratory of Life Analytical Chemistry, Institute of Chemistry, The Chinese Academy of Sciences (CAS), Beijing, China.
Zhang
Zipin
Z
Zhao
Lingzhi
L
Wang
Xiang
X
Yu
Ping
P
Su
Lei
L
Mao
Lanqun
L
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D001923:Brain Chemistry; D002384:Catalysis; D003199:Computer Systems; D004298:Dopamine / Q000032:analysis* / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation*; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D042845:Laccase / Q000737:chemistry*; D008297:Male; D009862:Online Systems; D010084:Oxidation-Reduction; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004800:Enzymes, Immobilized; D042845:Laccase; D004298:Dopamine