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Automatic optosensing device based on photo-induced fluorescence for determination of piceid in cocoa-containing products

Molina-García, Lucía, Ruiz-Medina, Antonio, Fernández-de Córdova, Maria Luisa
Analytical and bioanalytical chemistry 2011 v.399 no.2 pp. 965-972
chemoprevention, cocoa powder, cyclohexanes, ethanol, fluorescence, hydrodynamics, ionic strength, irradiation, lipoproteins, methanol, milk chocolate, neoplasms, resveratrol, solid phase extraction, toluene, ultraviolet radiation
Piceid (3,4′,5-trihydroxystilbene-3-β-d-glucoside) is a stilbene which occurs naturally in various families of plants and has been shown to protect lipoproteins from oxidative damage and to have cancer chemopreventive activity. This paper deals with the determination of piceid in cocoa-containing products by using photo-induced fluorescence and the aid of a multicommutated continuous-flow assembly which was provided with an on-line photoreactor. A strongly fluorescent photoproduct is generated from piceid when it is irradiated under UV light for 30 s, which is retained on Sephadex QAE A-25 and directly monitored on this active solid support at 257/382 nm (λ exc/λ em, respectively). The pre-concentration of the photoproduct of piceid on the solid support greatly improves both sensitivity and selectivity. The influence of different experimental parameters, both chemical (pH, ionic strength) and hydrodynamic (irradiation time, flow rate, photoreactor length, sampling time), was tested. The sample pre-treatment included delipidation with toluene and cyclohexane, stilbene extraction with ethanol/water (80:20, v/v) and clean-up by solid-phase extraction on C₁₈ cartridges and methanol/water (40:20, v/v) as eluting solution. This procedure allowed the elimination of the aglycon of piceid, resveratrol and other potential interfering species and a recovery of about a 90% piceid. The method was applied to the analysis of piceid in cocoa powder, dark chocolate and milk chocolate. The quantification limits were 1.4, 1.1 and 0.09 mg kg⁻¹, respectively. Relative standard deviations ranged from 1.8% to 3.1%. This is the first reported non-chromatographic method for determination of piceid in these foods.