Tyrosinase-Based Biosensor-A New Tool for Chlorogenic Acid Detection in Nutraceutical Formulations

The purpose of our research was to develop a new enzymatic biosensor, GPH-MnPc-Tyr/SPE, using as a support screen-printed carbon electrode (SPE) modified with graphene, manganese phthalocyanine, and tyrosinase, with the aim of developing sensitive detection of chlorogenic acid (CGA). To immobilise tyrosinase on the sensor surface, crosslinking with the glutaraldehyde technique was used, thus increasing the enzyme bioactivity on this electrode. The modified electrode has a great catalytic effect on the electrochemical redox of chlorogenic acid, compared to the simple, unmodified SPE. The peak current response of the biosensor for CGA was linear in the range of 0.1-10.48 mu M, obtaining a calibration curve using cyclic voltammetry (CV) and square-wave voltammetry (SWV). Subsequently, the detection limit (LOD) and the quantification limit (LOQ) were determined, obtaining low values, i.e., LOD = 1.40 x 10(-6) M; LOQ = 4.69 x 10(-6) M by cyclic voltammetry and LOD = 2.32 x 10(-7) M; LOQ = 7.74 x 10(-7) M, by square-wave voltammetry (SWV). These results demonstrate that the method is suitable for the detection of CGA in nutraceutical formulations. Therefore, GPH-MnPc-Tyr/SPE was used for the quantitative determination of CGA in three products, by means of cyclic voltammetry. The Folin-Ciocalteu spectrophotometric assay was used for the validation of the results, obtaining a good correlation between the voltammetric method and the spectrophotometric one, at a confidence level of 95%. Moreover, by means of the DPPH method, the antioxidant activity of the compound was determined, thus demonstrating the antioxidant effect of CGA in all nutraceuticals studied.

Automated summary

The purpose of this research was to develop a new enzymatic biosensor for sensitive detection of chlorogenic acid (CGA). The modified electrode showed a great catalytic effect on the electrochemical redox of CGA, leading to a linear peak current response. The detection limit and quantification limit were determined, and the method proved suitable for the detection of CGA in nutraceutical formulations. The correlation between the voltammetric method and the spectrophotometric one was good, and the antioxidant activity of CGA was demonstrated.