Monitoring the self-assembly of chitosan/glutaraldehyde/cysteamine/Au-colloid and the binding of human serum albumin with hesperidin

A new method for monitoring, in real-time, the self-assembly of chitosan/glutaraldehyde/cysteamine (CGC) on the gold surface and the immobilization of Au-colloid on CGC membrane with piezoelectric quartz crystal impedance (PQCI) are firstly proposed. Cyclic voltammogram and electrochemical impedance spectroscopy were also use to investigate the formation of Au-colloid/CGC. The viscosity-average molecular weight of chitosan used was firstly estimated as 16.8×105 with piezoelectric quartz crystal (PQC) sensor. On the basis of the analysis of the multi-dimensional information provided by PQCI, two stages existed in chitosan drying course: the frequency shift of the first stage was controlled by viscoelastic of the liquid, while the total frequency shift was due to mass change. The cross-link ratio of glutaraldehyde with chitosan was about 0.13, while for glutaraldehyde with cysteamine was about 0.217. PQCI also showed that the Au-colloid immobilization is a first-order reaction, while the HSA immobilization is a sum of two exponential functions, e.g., adsorption and re-arrangement. The association of the immobilized HSA with the purified hesperidin was monitored, and the association constant was estimated as 3.42 ml mg−1 by Scatchard analysis.