Flow electrochemical biosensors based on enzymatic porous reactor and tubular detector of silver solid amalgam

• Flow amperometric enzymatic biosensor was constructed.
• The biosensor is based on a reactor of a novel material – porous silver solid amalgam.
• Tubular amalgam detector was used for determination of decrease of O2 concentration.
• Covalent bonds amalgam−thiol−enzyme contributed to the sensor long-term stability.
• LOD of glucose was 0.01 mmol L−1 with RSD = 1.3% (n = 11).

A flow amperometric enzymatic biosensor for the determination of glucose was constructed. The biosensor consists of a flow reactor based on porous silver solid amalgam (AgSA) and a flow tubular detector based on compact AgSA. The preparation of the sensor and the determination of glucose occurred in three steps. First, a self-assembled monolayer of 11-mercaptoundecanoic acid (MUA) was formed at the porous surface of the reactor. Second, enzyme glucose oxidase (GOx) was covalently immobilized at MUA-layer using N-ethyl-N′-(3-dimethylaminopropyl) carboimide and N-hydroxysuccinimide chemistry. Finally, a decrease of oxygen concentration (directly proportional to the concentration of glucose) during enzymatic reaction was amperometrically measured on the tubular detector under flow injection conditions. The following parameters of glucose determination were optimized with respect to amperometric response: composition of the mobile phase, its concentration, the potential of detection and the flow rate. The calibration curve of glucose was linear in the concentration range of 0.02–0.80 mmol L−1 with detection limit of 0.01 mmol L−1. The content of glucose in the sample of honey was determined as 35.5 ± 1.0 mass % (number of the repeated measurements n = 7; standard deviation SD = 1.2%; relative standard deviation RSD = 3.2%) which corresponds well with the declared values. The tested biosensor proved good long-term stability (77% of the current response of glucose was retained after 35 days).

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