Rapid determination of phenylalanine and its related compounds in rumen fluid by high-performance liquid chromatography.

Abstract

A rapid method for the determination of phenylalanine (Phe), tyrosine (Tyr), benzoic acid (BZA), phenylacetic acid (PAA), phenyllactic acid (PLA), phenylpyruvic acid (PPY), phenylpropionic acid (PPR), and cinnamic acid (CNM) in goat rumen fluid was established by high-performance liquid chromatography (HPLC). The mobile phase used for isocratic elution was methanol-sodium acetate buffer (pH 6.5) (8:92, v/v). The compounds were monitored at 220 nm with a UV detector. A 5-microliters portion of the filtrated rumen fluid was analyzed and the analysis was completed within 20 min. The minimum detectable limits (microM) of these compounds were: 12 for Phe, 3 for Tyr, 3 for BZA, 9 for PAA, 12 for PLA, 15 for PPY, 20 for PPR, and 8 for CNM. The average contents of Phe, BZA, PAA, PLA, and PPR in the rumen fluid of three goats were 15.4, 73.7, 615.9, 51.1, and 39.9 microM before morning feeding, 17.0, 123.7, 650.4, 208.2, and 502.4 microM at 3 h after feeding, and 18.4, 124.2, 510.0, 129.9, and 178.5 microM at 6 h after feeding, respectively. Of these compounds PAA was present at the highest concentration both before and after feeding. The content of PPR extremely increased especially at 3 h after feeding. The other three compounds, i.e. Tyr, PPY, and CNM, were not detected in goat rumen fluid.