Aldehyde functionalized ionic liquid on electrochemically reduced graphene oxide as a versatile platform for covalent immobilization of biomolecules and biosensing

An aldehyde functionalized ionic liquid, (3-(3-formyl-4-hydroxybenzyl)-3-methylimidazolium hexafluorophosphate) (CHO-IL) has been employed herein as a multiple host platform for the covalent immobilization of mediator as well as enzyme. The CHO-IL was immobilized on electrochemically reduced graphene oxide (ERGO) through the pi-pi stacking of hydroxybenzyl and imidazolium groups with ERGO and subjected to further covalent attachment of Azure A (Azu-A) mediator or glucose oxidase (GOx) enzyme. Electroactive, water soluble organic dye Azu-A was effectively immobilized to the host IL through simple Schiff base reaction. Azu-A was rendered leak-free in the electrode setup and also responded well for the amperometric determination of H2O2 over a linear range of 0.03-1 mM with a detection limit and sensitivity of 11.5 mu M and 133.2 mu A mM(-1) cm(-2), respectively. Further, attempts were made to explore the CHO-IL platform for the covalent immobilization of GOx enzyme which served well in retaining the enzyme nativity, reactivity and stability. Under optimized conditions, mediatorless GOx biosensor developed based on direct electrochemistry has exhibited an impressive analytical signal towards glucose detection in the linear range of 0.05-2.4 mM with a detection limit and sensitivity of 17 mu M and 17.7 mu A mM(-1) cm(-2), respectively. The reliability of the proposed Azu-A based chemical sensor and GOx based biosensor towards the determination of H2O2 and glucose in the real samples have been demonstrated. The remarkable analytical parameters and long term stability of both the sensors could be envisioned as a result of facile immobilization platform and immobilization strategy.