As determined by high performance liquid chromatography, (-)-epicatechin concentrations among freshly harvested beans of verified genetic origin ranged from 21.89-43.27 mg/g of dry defatted samples. Fermented beans showed much lower concentrations (2-10 mg). During fermentation, a trend towards decrease in (-)-epicatechin content was observed. Commercial beans from areas with reputations for shipping well-fermented products contained lower levels of (-)-epicatechin than beans from regions where fermentation is less extensive.

In order to isolate, identify and characterize the microflora of cacao beans before, during and after fermentation and locate possible sources contributing to microbial contamination, cacao beans from the Centeno and San Louis Estates in Trinidad were investigated. Prior to fermentation, the interior and exterior of the pods, hands of employees, utensils, dried pulp material of the sweatboxes and finally fruitflies (Drosophila melanogaster) were studied microbiologically. At Centeno Estate, beans were sampled at 5, 45 and 90 cm depths at 8‐hr intervals for the first 72 hr and every 12 hr thereafter for 7 days. Sampling at San Louis Estate was carried out at 24 hr intervals for the same period. The changes in microbial population of the beans sampled at Centeno Estate ranged from 1.48 × 105/g at 0 hr to 4.1 × 105/g at the completion of the fermentation, whereas, at San Louis Estate they ranged from 6.8 × 105/g to 9.2 × 105/g during the same period. Taxonomical studies of isolates obtained during the fermentation period revealed the identification of 44 microorganisms at both Estates. Yeasts Zymomonas mobilis and several species of lactic acid organisms dominated the flora during the early stages of fermentation. As the fermentation progressed, these and other isolates were taken over by several species of genus Bacillus. Microbiological examination of dried and polished beans resulted in the identification of 22 organisms at Centeno Estate and 15 organisms at San Louis Estate

Glc of the pyrazine fraction from roasted cocoa beans yielded nine well resolved peaks which could be quantitated. When beans from several producing countries were roasted under identical conditions, pyrazines generated varied between 142 µg/ 100 g of beans and 698 µg/100 g. The potential for generating pyrazines was greatest in samples from countries where beans are traditionally fermented.

A sensitive high performance liquid chromatographic method for analysis of (-)epicatechin (3,3',4',5,7-pentahydroxyflavan) in cocoa beans is described. Bean samples were extracted in 80% acetone with subsequent sample clean-up on a Waters Associates Cta SEP-PAK. Separation of (-)-epicatechin was accomplished on a PBondapak Cta column using a mobile phase of water:methanol:acetic acid (87:8:5). (-)-Epicatechin was detected at 280 nm and quantified by comparing peak height of sample to those of standards. The method demonstrated excellent reproducibility and recoveries of added (-)epicatechin averaged over 90%. UV scans and mass spectrometric anlaysis of collected eluate from the chromatograph provided positive identification of the compound in cocoa bean extracts.

Glc of the silylated sugars fraction of cocoa beans revealed fructose, sorbose, glucose, sucrose, inositol, mannitol, a pentitol, and traces of two unidentified sugars. The same sugars were present in all samples, irrespective of geographic origin. Relative concentrations were, however, quite different, even among lots of the same type of bean. Most of these differences were attributed to harvesting, fermenting, and drying variables. Unfermented Sanchez beans were 1 % by wt sucrose, but fermented Bahia and Ghana beans averaged only 0.05 and 0.12% sucrose, respectively. Fermented varieties contained 2 to 16 times more fructose than glucose. The preferential consumption of the glucose moiety of sucrose during fermentation has implications important to the development of chocolate flavor during roasting. Results suggest that absorbed and occluded water-soluble constituents from the pulp contribute to the reducing sugars content of cocoa beans.A lthough cocoa beans contain only small amounts of L\ sugars, nonenzymatic browning reactions are, never--®theless, essential to the development of the typical aroma of chocolate. This is evidenced by the 30 pyrazines, 10 pyrroles, and 15 furans identified in the aroma fraction (van Elzakker and van Zutphen, 1961;Bailey et al., 1962;