Liquid chromatographic cleanup and determination of low levels of vitamin D3 in sheep plasma.
A liquid chromatographic (LC) method is described for the determination of vitamin D3 in sheep plasma. Samples are extracted by one of 2 different methods, depending on the concentration of vitamin D3. The samples are purified by using either a Sep-Pak silica cartridge or a small alumina column, followed by additional cleanup on a Metalsorb LC column. Final analysis was carried out on a 5 micron C18 column using a radial compression separation system with an acetonitrile-methanol solvent system. Vitamin D3 was completely resolved from any interfering compounds in the plasma; total run time was less than 15 min, using a variable wavelength detector set at 264 nm. The method was successfully applied to samples at levels of 1-10 ng added vitamin D3 mL sheep plasma, with recoveries in the range 90-97%.
D000818:Animals; D002762:Cholecalciferol / Q000097:blood* / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods; D012756:Sheep
D002762:Cholecalciferol