Laccase-Nafion Based Biosensor for the Determination of Polyphenolic Secondary Metabolites

Abstract

A laccase-based biosensor was developed by specific enzyme adsorption on screen-printed working electrodes of DROPSENS cells, and stabilized with Nafion 0.1% membrane. The electrode was characterized with respect to response time, sensitivity, linear range, detection limit, pH dependence, interferences, and long-term stability. The tested substrates were catechol, rosmarinic acid, caffeic acid, chlorogenic acid, and gallic acid. The optimized biosensor proved the following characteristic performances: the apparent Michaelis Menten calculated considering rosmarinic acid substrate 8.3 × 10-6 mol L-1 (r = 0.995, n = 6); the dynamic range of biosensor response for rosmarinic acid 7 × 10-7 - 1.5 × 10-6 mol L-1; the detection limit for rosmarinic acid 1.19 × 10-7 mol L-1 (RSD = 1.08%, n = 3). It was noticed that the biosensor reaches systematically 90% to 94.3% from the response obtained by LC-DAD-ESI-MS for real samples

Laccase immobilization on the electrode surface to design a biosensor for the detection of phenolic compound such as catechol

“…From Table 4, it is clear that sensitivity of our proposed biosensor for catechol, 706.7 mA L mol −1 , appeared higher than other offered biosensors [46][47][48][49][50][51][52]. Also detection limit is better (lower) in compare to some cases [48,51,52].…”

Monitoring of Rosmarinic Acid Accumulation in Sage Cell Cultures using Laccase Biosensor

“…Polyphenols from plants are either wild species or ‘ in vitro ’ synthesised secondary metabolites. The increase in the production yield may occur via chemical elicitation or as result of oxidative stress applied to ‘ in vitro ’ cultivated plants (Litescu et al ., 2010). Rosmarinic acid, for example, is rapidly accumulated during growth phase of sage plants subjected to oxidative light stress (Ruffoni et al ., ).…”

“…The necessity of simpler and user‐friendly techniques has led to these techniques being replaced by bioanalytical tools. During recent years many polyphenol oxidase‐based sensors have been reported in the literature for the determination of phenolic or polyphenolic compounds because of good selectivity, low cost and proven potential for miniaturisation and automation (Di Fusco et al ., ; Diaconu et al ., ; Litescu et al ., 2010).…”

“…In this context we report here the use of a previously developed simple laccase–nafion‐based biosensor developed by enzyme immobilising by adsorption on a working electrode surface and stabilisation with a nafion membrane (Litescu et al ., 2010). In the present work the focus is to provide information for very low concentrations of secondary polyphenolic metabolites and to monitor their accumulation during in vitro plant growth.…”

Voltammetric determination of polyphenolic content as rosmarinic acid equivalent in tea samples using pencil graphite electrodes

“…On the other hand, electrochemical methods of analysis are simpler, cheaper, more rapid and can be used either in turbid or colored solutions. Some biosensors using redox enzymes like peroxidase and laccase or/and tyrosinase have been developed for the evaluation of the TPC total polyphenolic content expressed as rosmarinic acid equivalents (Brondani et al 2011;Diaconu et al 2011;Litescu et al 2010;Eremia et al 2013) in different samples such as pharmaceuticals, olive oil, tea or plant extracts. RA was preferred as reference in these determinations, being redox recycled between the redox enzyme (enzymatic oxidation) and the electrode (electrochemical reduction).…”