SEPARATION OF POTENTIAL FLAVORING COMPOUNDS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

High-performance liquid chromatography separated successively and quantitatively the food flavoring agents pyrimidines, purines and nucleosides, followed by nucleotides, then by polyphenols and finally by pyrazines with a reversed-phase octadecylsilica (μBondapak C18) column and various proportions of methanol, water, acetic acid and tetrabutylammonium phosphate (PIC A). The polar solvent (solvent A) was water-acetic acid-PIC A (97.5:1.5:1.0) and the relatively non-polar solvent (solvent B) was methanol-acetic acid-PIC A (97.5:1.5:1.0). Purines, pyrimidines, and nucleosides were eluted with solvent A. Nucleotides were eluted with a mixture of solvents A and B (9:1). Polyphenols were separated with a gradient starting at 10% solvent B and finishing at 25% solvent B, and finally the pyrazines were removed successively from the column with a gradient starting at 25% solvent B and finishing at 45% solvent B. The resolution and reproducibility were excellent for more than 50 compounds. By this method beverages could be analyzed directly, without solvent extraction, for flavoring compounds. © 1979.