Vitamin D plays a vital role in skeletal metabolism, calcium homeostasis, and also in the functioning of the immune, cardiovascular, and reproductive systems. Vitamin D deficiency leads to rickets and osteomalacia and is also associated with breast and colorectal cancers, multiple sclerosis, dementia, rheumatoid arthritis, diabetes, Parkinson’s and Alzheimer’s diseases.
Consequently, laboratory testing of serum vitamin D levels has experienced phenomenal growth over the past several years. LC-MS/MS is currently the best technique available for the correct quantification of 25-Hydroxy Vitamin D3 and 25-Hydroxy Vitamin D2.Jasem method provides analysis of 25-Hydroxyvitamin D3, 25-Hydroxyvitamin D2 and 3-epi-25-Hydroxyvitamin D3 in serum samples using simple sample preparation technique without SPE, on-line SPE or evaporation.
25-OH Vitamin D3
25-OH Vitamin D2
Pipet 200 μL serum sample into a centrifuge tube
Add 100 μL of Reagent-1 and 25 μL of IS on sample and vortex for 15 seconds
Add 975 μL of Reagent-2 and vortex for an additional 15 seconds
Centrifuge at 4000 rpm for 5 minutes
Decant the supernatant into HPLC vial and inject into the system