DETERMINATION OF HIPPURIC ACID BY RP-HPLC USING TWO DIFFERENT ANALYTICAL COLUMNS – A SHORT REPORT
 
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Pol. J. Food Nutr. Sci. 2007;57(4):447–450
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ABSTRACT
Reversed-phase HPLC of liberated hippuric acid (HA) from an ACE assay in the presence of ACE inhibitory peptides derived from a crackling hydrolysate was conducted. The efficacies of two different analytical HPLC columns using identical mobile phases with an isocratic system were tested. Chromatograms revealed that the shorter C8 column (4.6 x 150 mm, 5 μm) was just as efficient as the longer C18 column (4.6 x 250 mm, 5 μm) in resolving liberated HA, but achieved this in a much shorter time (i.e., 3.67 cf 12.52 min). The presence of the crackling hydrolysate exhibited ACE-inhibiting activity by retarding the liberation of HA from the substrate hippuryl-L-histydyl-L-leucine (HHL) in a dose-dependent manner, and did not interfere with the chromatography. Hence, a quick reliable analytical methodology is at hand for the in vitro examination of various “bioactive peptide concoctions” for possible use in the development of functional foods.
ISSN:1230-0322