Simultaneous Measurements of Creatinine and Purine Derivatives in Ruminant's Urine Using Ion‐Pair HPLC

Abstract

A new and sensitive ion‐pair RP‐HPLC method was developed for concomitant measurement of the creatinine and purine derivatives (PD): allantoin, uric acid, and hypoxanthine using allopurinol as internal standard. Urine was chromatographed on a Symmetry C₁₈ column, using UV detection. Elution with a gradient of methanol in potassium phosphate buffer, pH = 4, containing 3 mM 1‐octane‐sulfonic acid, sodium salt yielded sharp, well‐resolved peaks within 25 min. Application of this method permits the precise determination of these compounds in sheep urine.

Keywords:

• Creatinine
• Purine derivatives