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HPTLC determination of epicatechin and procyanidin B2 in chocolate samples after matrix solid-phase dispersion

Flavonoids are natural polyphenolic antioxidants present in a wide range of plants, fruits and beverages. These compounds are of great interest because of their possible protective effects on human health in reducing the risk of chronic diseases such as cardiovascular diseases and cancers. Cocoa and chocolate are particularly rich in flavonoids with one class of those compounds being the flavan-3-ols, which included catechin and epicatechin and their oligomeric forms called procyanidins or proanthocyanidins. The extraction of these phenolic compounds from plants and other solid samples has been traditionally performed using solvent extraction or steam distillation techniques. Traditional methods of extraction are labour-intensive, time consuming and require large volumes of solvents. In the last years, several new methods of extraction such as solid-phase micro-extraction (SPME), supercritical fluid extraction (SFE), pressurized fluid extraction (PFE) and matrix solid-phase dispersion (MSPD) have been developed to overcome disadvantages of traditional methods. The aim of this work was to optimise matrix solid-phase dispersion (MSPD) of flavonoids epicatechin (EC) and procyanidin B2 (pB2) from chocolate samples. Extraction efficiency was investigated on C18 and Florisil sorbents with methanol, acetone, water, oxalic acid or their mixture as elution solvent. The goal of optimisation process was to find the sorbent and composition of elution solvent that gave the best recovery. A sample/sorbent ratio was 1:4. Extraction efficiency was determined by thin-layer chromatography using Milli Q water as mobile phase on HPTLC cellulose chromatographic plates (Merck, Germany) followed by videodensitometry in visible light after the derivatisation by spraying with DMACA (acidified 4-dimethylaminocinnamaldehyde in ethanol) reagent.

chocolate samples; matrix solid-phase dispersion; procyanidin B2; epicatechin

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