HPLC-MS-based design of experiments approach on cocoa roasting.

Modern statistical methods, such as the design of experiments and response surface methodology, are widely used to describe changes in multiparameter processes during the processing of food in both science and technology contexts. However, these approaches are described to a lesser degree in the case of cocoa roasting than other foods and processes. Our study aimed to use the design of experiments to establish a model of cocoa roasting for relevant flavor-related constituents. We have used HPLC-MS techniques to link standard process parameters with chemical compounds changing in concentration during cocoa roasting. Influence of time, temperature, the addition of water, acid, and base, on relative concentrations of procyanidin monomers, dimers, and trimers, an Amadori compound, and a peptide, was shown. High-quality models for each compound were established and validated, displaying good prediction accuracy. Such an approach could be used to optimize processing conditions for cocoa roasting in order to influence the concentration of certain chemical compounds, and in turn, improving the flavor of chocolate products.

Novel Amadori and Heyns compounds derived from short peptides found in dried cocoa beans.

Chemical transformations of Amadori compounds are responsible for the formation of aroma volatiles at the end of the Maillard reaction cascade, which in turn contributes to unique organoleptic characteristics of chocolate. A large amount of short peptides reported in fermented cocoa suggests the existence of a much larger variety of these flavor precursors than previously suspected. An HPLC-MS-MS study was performed on dried Malaysian cocoa beans to identify novel Amadori and Heyns compounds. In total, 34 species were found, including 26 previously unknown derived from di- and tripeptides. We illustrate how the structures were elucidated via tandem MS experiments, as well as present a comparative study on their relative quantities in samples coming from 11 countries of origin. There were significant differences between them, and discrimination was possible by principal component analysis based on Amadori content alone. However, the PCA separation could be a result of various post-harvest practices exerted among said countries.

Thermally-induced formation of taste-active 2,5-diketopiperazines from short-chain peptide precursors in cocoa.

2,5-diketopiperazines (DKPs) are cyclic dipeptides responsible for the specific bitter taste of cocoa formed during roasting. The 2,5-diketopiperazine and peptide composition of four different cocoa bean samples from different origins were studied using LC-MS techniques. 34 diketopiperazines were identified, of which 10 are newly reported in cocoa. Their formation was followed during two different roasting time-series using a zero-order and an alternative Prout-Tompkins solid-state kinetic models. The activation energies of diketopiperazine formation showed a distribution close to normal with individual values depending on the nature of the substituents. The relative concentrations of the DKPs were correlated with their putative peptide precursors in unroasted cocoa bean samples. The results showed a significant positive correlation, indicating that oligopeptides formed in cocoa bean fermentation are taste-precursors for bitter tasting diketopiperazines. Unexpectedly, for most diketopiperazines, a single major peptide precursor could be suggested.