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PubMed · 11439277

Chocolate.

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2001. Chocolate.. https://pubmed.ncbi.nlm.nih.gov/11439277/

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Improved high-performance liquid chromatography method to determine theobromine and caffeine in cocoa and cocoa products.

At present, the commonly used HPLC method for the analysis of caffeine and theobromine contents in aqueous cocoa extracts employs direct application of the extracts on the column. This practice gradually reduces the efficiency of the column and shortens its life. Also, this method gives inflated values due to interfering substances and difficulty in achieving baseline resolution. In the improved method, the interfering cocoa pigments are effectively removed by passing the aqueous extract through a Sep-pak C(18) cartridge. Subsequent injection on a C(18) reverse-phase column employing acetonitrile and water (20:80) as the mobile phase reduces the analysis time without affecting either resolution of the peak or the accuracy of caffeine and theobromine determination or achieving baseline resolution. Therefore, this method is ideally suited for rapid routine analysis of cocoa and its products.

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Dark chocolate tablets were manufactured using 100% crystalline sucrose, 50% crystalline and 50% amorphous sucrose, and 100% amorphous sucrose. The physical state of sucrose was determined by differential scanning calorimetry (DSC) and X-ray diffraction. DSC scans of dark chocolate samples containing amorphous sucrose were characterized by a glass transition at 63 degrees C, a sucrose crystallization peak at 105 degrees C, and a melting endotherm at 188 degrees C. Independent of the amount of amorphous or crystalline sucrose used for the preparation of dark chocolate, all final chocolate products provided a single melting endotherm at 188 degrees C and a crystalline X-ray diffraction pattern. These results indicated that sucrose crystallized during production of dark chocolate and that no amorphous sucrose was present in the final chocolate products.

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