A RAPID PROCEDURE FOR PURINE MEASUREMENT AND ITS USE FOR ESTIMATING NET RUMINAL PROTEIN SYNTHESIS

Abstract

A rapid method for separation and quantitation of purines was applied to ruminal and intestinal digesta for estimating net microbial protein synthesis in the rumen. The procedure combines standard literature methods for hydrolysis of nucleotides by perchloric acid followed by precipitation of free purines with silver nitrate to separate the purines from interfering compounds. Acid resolubilized purines were quantitated spectrophotometrically at 260 nm. Microbial protein was estimated by the ratio of purines to N of isolated bacteria. The procedure is rapid, simple, precise and not costly. Duodenal passage of microbial N estimated by this procedure for steers fed semipurified and purified diets containing no protein was highly correlated (R 2 = 0.98; P < 0.01) with duodenal passage of tungstic acid precipitable N. Results indicate that purines may be useful as a marker for quantitating microbial protein. Key words: Purine, RNA, DNA, microbial protein