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

Martin Perkins

13th December 2017

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 (25-OH-D2) and 25-Hydroxyvitamin D3 (25-OH-D3), respectively. These metabolites are routinely measured for diagnostic assessment of Vitamin D related diseases. For some laboratories, a large number of samples require analysis in a very short space of time and the manual procedure is time consuming and repetitive. A fully automated solution was developed featuring protein crash step, solid phase extraction (SPE) and LC-MS/MS analysis. The method fully separates 25-OH-D3/D2 from the inactive form Epi-25-OH-D3/D2. The comprehensive prep ahead capability in Maestro software allows the system to fully process and analyse 96 samples in just under seventeen hours. This offers significant advantages when compared to a standard manual approach requiring sample preparation with subsequent LC-MS analysis.