Cocoa flavanols and procyanidins (CFs), natural dietary bioactives, have been studied extensively over the past two decades for their potential health benefits. Research on their safety and efficacy is critically dependent upon on the ability to reliably characeterize the research materials that are utilized, and with growing consumer availability of CF-based products, reliable methods for the detection of potential adulteration are of increasing importance. This research focused on the development of a high performance liquid chromatography-tandem mass spectrometry method (HPLC-MS2) using primary standards and13C-labelled procyanidins as internal standards.
Fig: Structure of (−)-epicatechin and (−)-epicatechin oligomers (C4 → C8 B-type procyanidin) with degree of polymerization up to five (DP2–5), red dots represent carbon that are labelled with 13C on synthetic procyanidin standards. (structures drawn with ChemDraw Prime v17.1).
The ability of MS2detection to discriminate A- and B-type procyanidins was demonstrated. Method performances were validated for degrees of polymerization up to four in seven model food matrices. Accuracy ranged from 90.9 to 125.4% and precision was < 10% at lower concentrations. Finally, the method was applied to cocoa-based samples and compared to the AOAC 2020.05 analytical protocol, supporting the use of NIST 8403 as reference material for HPLC-MS2 analysis.
Bussy, U., Olanrewaju, Y., Crozier, A. et al. Development and validation of HPLC-MS2 methodology for the accurate determination of C4–C8 B-type flavanols and procyanidins. Sci Rep 11, 14761 (2021). https://doi.org/10.1038/s41598-021-93993-0