An EQCM study on the interaction of heparin with the charge-transfer complex generated during o-tolidine electrooxidation: A biosensing mode with a dynamically renewed surface

The electrooxidation of o-tolidine (oTD) was investigated via the electrochemical quartz crystal microbalance (EQCM) technique. The formation and breakage of the poorly soluble charge-transfer complex (CTC) occurred during the redox switching of oTD, and the CTC precipitation on and its removal from the electrode surface led to a V-shaped frequency response to the cyclic voltammetric switching of oTD. The V-shaped frequency response to the redox switching of the CTC/oTD “couple” and the electrode-collection efficiency of the CTC precipitate were notably enhanced by the introduction of sodium heparin due to the formation of the CTC–heparin adduct as reported here for the first time. FTIR and UV-Vis characterizations also supported the interaction between the CTC and heparin. The molar ratio of the positively charged CTC to negatively charged heparin of the adduct was estimated here to be between 31.5 and 36.5, being close to the anticipated value, 37.5, for the full electrical neutralization in the adduct. An EQCM-based biosensor featured by a dynamically renewed surface of the detection electrode was proposed for heparin assay, with a limit of detection of 18.5 nM (S/N = 3) in pH 6.0 Britton–Robinson buffer solution containing a 10-fold diluted blood serum. This method is convenient in operation and highly free from the interference from coexisting substances including proteins. The new and intriguing biosensing concept based on the labile CTC-“target” adduct is featured by a dynamically renewable and regenerable surface of the detection electrode, and it is highly recommended for wide biosensing and electroanalytical applications.