Abstract
Benzoic acid (BA) of natural origin in fermented milk is known to be derived from hippuric acid (HA) in raw milk. Since it is forbidden to fortify fermented milk with BA, BA in fermented and HA in raw milk should be precisely determined in order to assess the source of BA.Our method of simultaneous analysis for both organic acids consists of extracting with acetone, making the extract alkaline with sodium bicarbonate solution, concentrating the mixture, and injecting a sample into a high performance liquid chromatography (HPLC) apparatus. HPLC was performed on a Zorbax ODS column, with 2% acetic acid-acetonitrile (78:22) as the mobile phase and o-methylhippuric acid as an internal standard. The detection limits of BA and HA were each 1ng, and the average recoveries of BA and HA added to milk at levels of 5-40ppm were not less than 98%.
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