We have developed a high-performance liquid chromatography (HPLC) method for assay of hippurate in plasma of patients with renal failure. Hippurate accounts, in part, for the impaired binding of drugs and metabolites to albumin and may cause other disorders in azotemic patients. The method is precise, accurate and reproducible. Among 25 patients with acute and chronic renal failure having serum creatinine in the range of 2.9-43 mg/dl (256-3,801 μmol/l), plasma hippurate ranged from 0.11 to 16.2 mg/dl (6.1-904 μmol/l). Hippurate concentration correlated moderately closely with plasma creatinine, urea and anion gap. Its curvilinear relation to the reciprocal of serum creatinine indicated a proportional decline of GFR and tubular function or the accumulation of inhibitors of the proximal tubular anion secretory pathway. The method should be useful for further studies of abnormal albumin binding as well as other disorders in azotemic patients.