Unsaturated vitamin b(12) binding capacity in human and ruminant blood serum - a comparison of techniques including a new technique by high performance liquid chromatography

A new technique by High Performance Liquid Chromatography (HPLC-gel permeation) shows promise as a tool to separate and quantitate the Unsaturated Vitamin B(12) Binding Capacity (UBSC) of the individual Vitamin B(12) binders in blood serum. This method, although not as rapid as protein-coated charcoal or cellulose separation techniques, is more applicable for use with large numbers of samples than gel filtration. The use of a radioactivity detector to monitor the eluant from the column permitted automation of the method. Comparable results for UBBC and for the UBBC of individual binders were obtained when samples were analyzed by gel filtration and HPLC. The HPLC method proved suitably precise and the recovery of added cyanocobalamin was acceptable. It is proposed that HPLC be the method of choice for measurement of the USBC of binders of Vitamin B(12) in blood serum.