Urinary Protein and Albumin Determinations by High-Performance Gel-Permeation Chromatography

Abstract Sensitive high-performance gel-permeation chromatographic protein assay method was developed by using non-ionic detergent of Brij-58 which was UV transparent as compared to Nonidet P-40. A column (70 × 8.0 mm I.D.) packed with Develosil 100 Diol-5 (pore size 10 nm) was used for the protein determination. A commercially available column (300 × 8.0 mm I.D.) packed with Develosil 300 Diol-5 (pore size 30 nm) was used for the urinary albumin determination. Eluent used was a 0.1 M sodium phosphate buffer (pH 5.6) containing 0.3 M sodium chloride, 1% (v/v) Brij-58, 50% (v/v) glycerol. Flow-rates for protein assay and albumin assay were 1.0 and 0.5 ml/min, repectively. Proteins were eluted at the position of the exclusion limit in the case of protein assay (100 Diol-5). Albumin was eluted from 300 Diol-5 column as a symmetric peak. Bovine serum albumin (BSA) was used as an external standard for both protein and albumin assays. Analysis times were within four min for protein assay, and 32 min for albumin...