Improved biosensor for glucose based on glucose oxidase-immobilized silk fibroin membrane

Based on glucose oxidase-immobilized silk fibroin membrane and oxygen electrode, the authors have developed an amperometric glucose sensor in flow-injection analysis. After the sensor was improved by the configuration of oxygen electrode and a temperature control system was added to the electrode body, its sensitivity, analytical precision, and stability were enhanced greatly. The authors first introduced a tailing inhibitor-ion pair reagent into a buffer system in the biosensor so as to eliminate all interference from hemacyte, macromolecules, and small mol wt charged species besides electroactive specie ascorbate in complex matrices. A considerably serious tailing of the biosamples, such as whole blood, plasma, serum, or urine on the sensor, based on enzyme electrode, entirely disappeared, their response times were shortened, and base lines became more smooth and stable. The glucose sensor has a broad range of linear response for glucose (up to 25.0 mmol/L) and a good correlation (γ = 0.999) under conditions of control temperature 32.0°C and 1.6 mL/min 0.02 mol/L phosphate buffer containing 0.5% tailing inhibitor (v/v). Recoveries of glucose in these biosamples are within the range of 93.71–105.88%, and its repeatabilities for determining glucose, repeated 100 times, human blood dilution 125 times, and serum 128 times, are 1.81,2.48, and 2.91% (RSD), respectively. The correlation analysis for 200 serum samples showed that the correlation (γ) is 0.9934 between the glucose sensor and Worthington method for determining serum glucose used conventionally in a hospital laboratory. Moreover, the enzyme membrane used in the biosensor can be stored for a long time (over 2 yr) and measured repeatedly over 1000 times for biosamples. The glucose sensor is capable of detecting over 60 biosamples/hr.