Protein determination by high-performance gel-permeation chromatography: applications to human pancreatic juice, human bile and tissue homogenate

A high-performance gel-permeation chromatography (HPGPC) method to determin e the proteins of human pancreatic juice, bile, and tissue homogenate has b een developed. A diol-type silica gel column (35X8 mm I.D., 5 nm average po re diameter) was used under a column temperature of 8 degreesC. The eluent was acidic phosphate buffer with a high concentration of sodium chloride, n onionic detergent of polyoxyethylene (20) cetyl ether (Brij 58), glycerol a nd 2-propanol. The UV wavelength used for the protein detection was 210 nm. Analytical time was within 3.5 min. Good correlation coefficients were obt ained with this HPLC method at a column temperature of 8 degreesC and a spe ctrophotometric bicinchoninic acid (BCA) method. A photometric pyrogallol-r ed molybdate complex method was found to correlate well with this HPLC meth od and with the BCA method only for tissue homogenate. Since this HPGPC pro tein assay method is simple, convenient, rapid, reproducible, and reliable, it is expected to be generally applicable to clinical and also to biochemi cal research. (C) 2001 Elsevier Science B.V. All rights reserved.