FSCN 4613 Lecture Notes - Lecture 5: Solid Phase Extraction, Cholecalciferol, Ethyl Acetate
23 Mar 2020
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25-hydroxyvitamin d3 (calcidiol) 1, 25-dihydroxyvitamin (calcitriol) By enzyme 1-hydroxylase (located in the kidney) Form that most accurately reflects vitamin d3 status. Concentrations of 1,25-dioh-d3 usually do not change until a deficiency is severe, due to the tight regulation of its synthesis. You will extract your lipids using solid-phase extraction (spe) Therefore, there is a mobile phase and a stationary phase. Used to separate compounds based on their relative affinities for the solid phase. Used for sample purification and concentration, not for quantification and identification. Extraction of 25-oh-d3 from your serum by solid-phase extraction. Add a precipitation reagent (acetonitrile) to precipitate proteins. Centrifuge to precipitate proteins, then transfer supernatant to a clean tube. Add distilled water to your sample, then wrap w/ aluminum foil to prevent photodegradation. Process your sample through the solid phase extraction column to partially purify your sample. To the dried tube, add exactly 150 microliters of ethyl acetate: acetonitrile, vortex well, add 100 microliters water, vortex again.
