Waveform Optimization for the In Vitro Detection of Caffeic Acid by Fast-Scan Cyclic Voltammetry.

Abstract

Caffeic acid is a polyphenol of critical importance in plants, involved in a variety of physiological processes including lignin formation, cellular growth, stress response, and external signaling. This small molecule also acts as a powerful antioxidant and thus has therapeutic potential for a variety of health conditions. Traditional methods of detecting caffeic acid lack appropriate temporal resolution to monitor real time concentration changes on a subsecond time scale with nM detection limits. Here we report on the first usage of fast-scan cyclic voltammetry with carbon fiber microelectrodes for the detection of caffeic acid. Through the use of flow injection analysis, the optimal waveform for its detection under acidic conditions at a scan rate of 400 V/s was determined to be sawtooth-shaped, from 0 to 1.4 to -0.4 to 0 V. Signal was linear with concentration up to 1 μM with a sensitivity of 44.8 ± 1.3 nA/μM and a detection limit of 2.3 ± 0.2 nM. The stability of its detection was exceptional, with an average of 0.96% relative standard deviation across 32 consecutive injections. This waveform was also successful in detecting other catechol-based plant antioxidants including 5-chlorogenic acid, oleuropein, rosmarinic acid, chicoric acid, and caffeic acid phenethyl ester. Finally, we show the successful use of fast-scan cyclic voltammetry in monitoring the degradation of caffeic acid by polyphenol oxidase on a subsecond time scale via a novel modification of a Ramsson cell. This work demonstrates that fast-scan cyclic voltammetry can be used to successfully monitor real-time dynamic changes in the concentrations of catechol-containing plant polyphenols.