Separation of guinea-pig pancreatic juice proteins by reversed-phase high-performance liquid chromatography

Using a Protesil 300 octyl reversed-phase column with a multistage water-acetonitrile solvent gradient system, it was possible to obtain a well-resolved separation of the nine major proteins present in guinea-pig pancreatic juice. The protein present in each peak of the pancreatic juice chromatogram could only be identified by molecular weight analysis as the acetonitrile denaturated the enzymes and altered their isoelectric points. However, using sodium dodecyl sulphate gel electrophoresis the protein fractions obtained by high-performance liquid chromatography were characterised. Preliminary work has indicated that this system may be capable of separating other complex biological protein mixtures, i.e., saliva.