Influence of Aerobic and Anaerobic Moist Incubation on Selected Nonvolatile Constituents─Comparison to Traditionally Fermented Cocoa Beans.

Recently, moist incubation has been proposed as an alternative postharvest processing method for cocoa beans. During this treatment, unfermented and dried cocoa nibs are rehydrated with a lactic acid solution containing ethanol and subsequently incubated for 72 h at 45 °C before drying. Previous studies focused on the aroma formation during this treatment and the further processing of chocolate. The current study focused on the influence of aerobic and anaerobic moist incubation on selected nonvolatile components in comparison with the unfermented raw material and traditionally fermented cocoa. Total phenolic content and total flavan-3-ol content, contents of (+)-catechin, (-)-epicatechin, procyanidins B2 and C1, cinnamtannin A2, methylxanthines (theobromine and caffeine), contents of sugars (sucrose, d-glucose, and d-fructose) and free amino acids (17 proteinogenic amino acids) were determined. The fermentation index was also evaluated. The aerobically incubated and fermented cocoa showed low levels of phenolic compounds in comparison to the unfermented cocoa and the anaerobically incubated cocoa. The level of methylxanthines was unaffected by all treatments. The contents of reducing sugars were more than 2-fold higher after both incubation treatments compared to fermentation. The level of free amino acids liberated was highest after anaerobic incubation followed by fermentation and aerobic incubation. The aerobically incubated cocoa showed the highest FI, while the anaerobically incubated cocoa may be considered under-fermented (FI <1.0). Statistical analysis (ANOVA) showed significant differences between all treatments, which was verified by principal component analysis.

Comparison of the Aroma Composition and Sensory Properties of Dark Chocolates Made with Moist Incubated and Fermented Cocoa Beans.

In a previous investigation, "moist incubation" was described as a novel postharvest treatment for cocoa and the aroma composition of the resulting cocoa nibs was compared to unfermented and fermented cocoa nibs. For this treatment, unfermented and dried nibs are rehydrated with an aqueous solution containing lactic acid and ethanol to adjust the pH value and are subsequently incubated at 45 °C under aerobic conditions for 72 h before drying. The aim of the present study was to investigate the sensory properties and aroma composition of dark chocolates made of these materials after roasting. Therefore, gas chromatography-olfactometry (GC-O) in combination with aroma extract dilution analysis (AEDA), quantitation with isotopically labeled standards, odor activity value (OAV) determination, and sensory analysis were performed. The three different chocolates had distinct sensory and OAV profiles. The sensory profiles showed a higher intensity of fruity aroma notes and a lower intensity of bitterness and astringency in the chocolate made with the moist incubated cocoa, while the chocolate made of fermented cocoa reached higher scores in the roasty aroma notes. Furthermore, higher OAVs were determined for the Strecker aldehydes in the chocolate made of the moist incubated cocoa, whereas higher OAVs for the pyrazines and the acids were detected in the chocolate made of fermented cocoa. In contrast, the chocolate produced with the unfermented cocoa showed low cocoa specific aroma notes and high levels of astringency and bitterness. The detected differences reveal interesting insights into the influence of different postharvest treatments on the resulting aroma composition in the final chocolate. Furthermore, the alternative postharvest treatment was demonstrated to result in chocolates with a pleasant sensory profile.

Novel Time- and Location-Independent Postharvest Treatment of Cocoa Beans: Investigations on the Aroma Formation during "Moist Incubation" of Unfermented and Dried Cocoa Nibs and Comparison to Traditional Fermentation.

The aroma properties of cocoa nibs obtained by applying a novel postharvest treatment were investigated using methods of the molecular sensory science approach, i.e., solvent extraction and solvent-assisted flavor evaporation, aroma extract dilution analysis (AEDA), stable isotope dilution analysis, calculation of odor activity values (OAVs), and orthonasal sensory evaluation; those properties were then compared to the unfermented and dried raw material and a traditionally fermented sample of the same harvest. For the treatment, unfermented and dried cocoa nibs were, first, rehydrated with lactic acid and ethanol solution to adjust the pH value to 5.1 and, second, incubated under aerobic conditions for 72 h at 45 °C and subsequently dried. This treatment was used to induce enzymatic reactions within the cotyledon matrix, which also occur inside the bean during microbial fermentation of the surrounding fruit pulp. The results of the AEDA showed that many of the key aroma compounds found in fermented and dried cocoa increased during the incubation treatment. Especially some "fruity" esters were found with an equal or even higher flavor dilution (FD) factor in the incubated sample compared to the fermented sample, whereas the fermented sample showed high FD factors for "pungent, sour" and "sweaty" acids, such as acetic acid and 2- and 3-methylbutanoic acids. The quantitative data and calculated OAVs for the samples supported the findings of the AEDA, underlining the potential of this approach as a controllable and reproducible alternative postharvest treatment.

Investigations on the Aroma of Cocoa Pulp ( Theobroma cacao L.) and Its Influence on the Odor of Fermented Cocoa Beans.

The odor-active constituents of cocoa pulp have been analyzed by aroma extract dilution analysis (AEDA) for the first time. Pulps of three different cocoa varieties have been investigated. The variety CCN51 showed low flavor intensities, in terms of flavor dilution (FD) factors, in comparison to varieties FSV41 and UF564, for which floral and fruity notes were detected in higher intensities. To gain first insights on a molecular level of how the cocoa pulp odorants affected the odor quality of cocoa beans during fermentation, quantitative measurements of selected aroma compounds were conducted in pulp and bean at different time points of the fermentation. The results showed significantly higher concentrations of 2-phenylethanol and 3-methylbutyl acetate in pulp than in the bean during the different time steps of the fermentation, whereas the reverse could be observed for the odorants linalool and 2-methoxyphenol. The findings of this study constitute a basis for further investigations on the aroma formation of cocoa during fermentation.