Marcelo Ricardo Romero

Amperometric biosensor for direct blood lactate detection.

An amperometric sensor for lactate quantification is presented. The developed biosensor requires only 0.2 U of lactate oxidase, which is immobilized in a mucin/albumin hydrogel matrix. By protecting the platinum surface with a Nafion membrane, typical interference related to negatively charged species such as ascorbic acid has been minimized to practically undetectable levels. Electrochemical properties associated with the Nafion membrane are assessed as a function of Nafion concentration. In a phosphate buffer solution of pH 7.0, linear dependence of the catalytic current upon lactate bulk concentration was obtained between 2 and approximately 1000 microM. A detection limit of 0.8 microM can be calculated considering 3 times the standard deviation of the blank signal divided by the sensitivity of the sensor. The lactate biosensor presents remarkable operational stability and sensitivity (0.537 +/- 0.007) mA.M(-1), where the error is the standard deviation of the slope calculated from the linear regression of the calibration curve of a fresh biosensor. In this regard, the sensor keeps practically the same sensitivity for 5 months, while the linear range decreases until an upper value of 0.8 mM is reached. Assays performed with whole blood samples spiked with 100 microM lactate gave (89 +/- 6)% of recovery.

Mucin and carbon nanotube-based biosensor for detection of glucose in human plasma

This work reports an amperometric enzyme-electrode prepared with glucose oxidase, which have been immobilized by a cross-linking step with glutaraldehyde in a mixture containing albumin and a novel carbon nanotubes-mucin composite (CNT-muc). The obtained hydrogel matrix was trapped between two polycarbonate membranes and then fixed at the surface of a Pt working electrode. The developed biosensor was optimized by evaluating different compositions and the analytical properties of an enzymatic matrix with CNT-muc. Then, the performance of the resulting enzymatic matrix was evaluated for direct glucose quantification in human blood plasma. The novel CNT-muc composite provided a sensitivity of 0.44 ± 0.01 mA M-1 and a response time of 28 ± 2 s. These values were respectively 20% higher and 40% shorter than those obtained with a sandwich-type biosensor prepared without CNT. Additionally, CNT-muc based biosensor exhibited more than 3 orders of magnitude of linear dynamic calibration range and a detection limit of 3 μM. The short-term and long-term stabilities of the biosensors were also examined and excellent results were obtained through successive experiments performed within the first 60 days from their preparation. Finally, the storage stability was remarkable during the first 300 days.