Vitamin D, along with calcium, promotes bone growth in children and aids prevention of osteoporosis in adults. It is present in two forms, Vitamin D3 and Vitamin D2, which are both metabolised in the liver to form 25-Hydroxyvitamin D2 (25OH-D2) and 25-hydroxyvitamin D3 (25OH-D3), respectively. These metabolites are routinely measured for diagnostic assessment of Vitamin D related diseases. A fully automated solution was developed featuring protein crash step, solid phase extraction (SPE) and LC-MS-MS analysis. The method fully separates 25OH-D3/D2 from the inactive form Epi-25OHD3/D2. The comprehensive prep ahead capability in Maestro software allows the system to process 96 samples in just under seventeen hours.
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Fully Automated Sample Preparation and Analysis of 25-OH-Vitamin D3/D2 and 3-Epi-25-OH Vitamin D3/D2 in Serum using LC-MS:MS