Development of a method for the separation of corrinoids in ovine tissues by HPLC

Biomed Chromatogr. 2005 May;19(4):329-33. doi: 10.1002/bmc.444.

Abstract

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.

MeSH terms

  • Animals
  • Body Fluids / chemistry
  • Chromatography, High Pressure Liquid / methods*
  • Cobalt Radioisotopes
  • Cobamides / isolation & purification
  • Corrinoids / blood
  • Corrinoids / isolation & purification*
  • Female
  • Intestines / chemistry
  • Liver / chemistry
  • Milk / chemistry
  • Radioisotope Dilution Technique
  • Reproducibility of Results
  • Sheep
  • Vitamin B 12 / analogs & derivatives*
  • Vitamin B 12 / isolation & purification

Substances

  • Cobalt Radioisotopes
  • Cobamides
  • Corrinoids
  • mecobalamin
  • cobamamide
  • Vitamin B 12