Quantitative analysis of proanthocyanidins in cocoa using cysteamine-induced thiolysis and reversed-phase UPLC.

The thiolysis of B-type proanthocyanidins in cocoa by cysteamine was evaluated and optimized for its application in cocoa proanthocyanidin quantification. Four thiolysis products consisting of epicatechin, catechin, and their thioethers formed with cysteamine were separated and characterized by reversed-phase UPLC with photo diode array (PDA) detection and high-resolution mass spectrometry (HRMS). A thiolysis time of 20 min under 60 °C temperature was determined as the optimal condition for cocoa proanthocyanidin depolymerization. The optimized thiolysis condition was applied to four cocoa bean samples for proanthocyanidin quantification, using commercially available procyanidin B2 dimer as a reference standard. Satisfactory linearity and quantification and detection limits were achieved for the calibration curves, and proanthocyanidin contents determined by thiolysis were found to be higher than those determined by a published method based on normal-phase HPLC with fluorescence detection. Results in this study suggest promising application potential of cysteamine as an odorless thiolysis agent in routine quantitative analysis of B-type proanthocyanidins. Graphical abstract.