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

Abstract

A stable, sensitive and simple biosensor based on laccase-nafion was used for monitoring the total polyphenolic content from two in vitro cultivated plants. The biosensor response was free of electrochemical interferences and of possible interferences from growth media constituents, demonstrating a high sensitivity for rosmarinic acid determination in cell culture suspensions.

Antioxidant Capacity Determination in Plants and Plant‐Derived Products: A Review

“…The content of phenolics in the analysed samples was assessed through interpolation of the peak area using the calibration curve developed per reference to the rosmarinic acid peak and retention time. The results obtained, as rosmarinic acid equivalent content, ranged from 103 ± 2  µ g/g fresh material for S. maxima to 174 ± 2  µ g/g fresh material for Salvia verde , with a limit of detection of 3.4 × 10 −7  mol L −1 [110]. …”

“…The results, as rosmarinic acid equivalent content (chosen as standard, as it has been identified as the major phenolic in the samples analysed, by chromatography and MS screening), ranged from 97.8 ± 8.2  µ g/g fresh material for Salvia maxima to 162.2 ± 11.3  µ g/g fresh material for Salvia verde , with a limit of detection of 4.2 × 10 −7  M. The biosensor retains more than 85% of its initial analytical response up to 90 days. The Michaelis-Menten kinetic apparent constant of 8.3 × 10 −6  M revealed the enzyme affinity for the substrate [110]. …”

Recent Prospects of Carbonaceous Nanomaterials-Based Laccase Biosensor for Electrochemical Detection of Phenolic Compounds

“…Rosmarinic acid is the chief phenol present on both herbs [ 178 ]. Eremia et al determined rosmarinic acid from in vitro saliva culture with Lac–Nafion-based biosensors [ 179 ].…”

Electrochemical Methods and (Bio) Sensors for Rosmarinic Acid Investigation

“…RA has multiple biological and pharmaceutical activities: antioxidant, anti-inflammatory, anti-mutagenic, antibacterial, anti-tumor, hepato-protective and anti-hepatitis, cardio-protective, antiviral, anti-carcinogenic, anti-convulsant, immunomodulatory, anti-metastatic, anti-angiogenic, astringent and anti-allergic [16,17,[29][30][31][32]. It also helps the fertilization capacity increase [13] and the cognitive efficiency enhancement (neuroprotective activity) [19].…”

“…Electrochemical methods are eco-friendly because: (i) they need a reduced number of reagents (mainly non-toxic supporting electrolytes, e.g., phosphate buffer [7,21,[29][30][31]33], McIlvaine buffer [8], acetate buffer solution [16,31], etc.) and small sample volumes (e.g., approximately 10 mL in conventional electrochemical cells [16,[29][30][31], 1 mL using one compartment cells [17] or µL/few drops when using screen printed electrodes-SPE [7,32]), generating thus insignificant waste amounts; (ii) many analytes (e.g., polyphenols and especially, in the present discussion, RA) are electroactive and they can be thus detected directly, without previous derivatization, and (iii) most often, even for mixtures or complex matrices, the sample preparation involves only basic operations, such as filtration (e.g., RA determination in Rosmarinus officinalis L. and Melissa officinalis herbs, where the samples were mixed with ultrapure water, heated at 50 • C, filtered and aliquots of the supernatant were voltammetrically analyzed without any dilution [13]), extraction (with water:ethanol mixtures from sage [8], from Prunella vulgaris using ethanol [33] and with methanol from spices [43]) and dilution, without the requirement of tedious, time and reagents consuming steps.…”