Development of a method for the separation of corrinoids in ovine tissues by HPLC.
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.
Kelly
R J
RJ
Keratec Limited, 1375 Springs Road, Lincoln, Canterbury 8152, Private Bag 4749, Christchurch, New Zealand.
Gruner
T M
TM
Sykes
A R
AR
D000818:Animals; D001826:Body Fluids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003037:Cobalt Radioisotopes; D003038:Cobamides / Q000302:isolation & purification; D045728:Corrinoids / Q000097:blood / Q000302:isolation & purification*; D005260:Female; D007422:Intestines / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008892:Milk / Q000737:chemistry; D011865:Radioisotope Dilution Technique; D015203:Reproducibility of Results; D012756:Sheep; D014805:Vitamin B 12 / Q000031:analogs & derivatives* / Q000302:isolation & purification
D003037:Cobalt Radioisotopes; D003038:Cobamides; D045728:Corrinoids; C019476:mecobalamin; C000913:cobamamide; D014805:Vitamin B 12