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Development of a method for the separation of corrinoids in ovine tissues by HPLC
Author(s) -
Kelly R. J.,
Gruner T. M.,
Sykes A. R.
Publication year - 2005
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.444
Subject(s) - methylcobalamin , chemistry , hydroxocobalamin , chromatography , high performance liquid chromatography , adenosylcobalamin , sodium acetate , cobalamin , cyanocobalamin , biochemistry , vitamin b12
A method has been developed using a combination of high‐performance liquid chromatography (HPLC) and a radioisotope dilution assay (RIDA) to routinely estimate the distribution of corrinoids (the cobalamins hydroxocobalamin, methylcobalamin and 5′‐deoxyadenosylcobalamin, and cobalamin analogues) in liver, plasma, milk, intestinal fluid and faeces. Corrinoids were extracted with a sodium acetate buffer, separated by HPLC and quantified by RIDA. Recoveries of corrinoids were 29% for hydroxocobalamin, 50% for 5′‐deoxyadenosylcobalamin and 64% for methylcobalamin. The method allows the routine analysis of many samples and maintains good standards of precision. Copyright © 2005 John Wiley & Sons, Ltd.