Automated Sample Preparation of 25-OH-Vitamin D3/D2 from Serum with the AC Extraction Plate and the Tecan Freedom EVO® for the quantification by LC-MS/MS

The current methods for sample preparation include protein precipitation (‘protein crash’), solvent extraction such as liquid-liquid extraction (LLE) and solid-phase extraction (SPE). 

However, all these methods require at least one labor intensive and time-consuming step such as filtration, centrifugation or solvent evaporation. In addition, these steps do not allow straightforward automation of the sample preparation process which is highly desirable in order to cope with the increasing Vitamin D sample load experienced by clinical laboratories. 

The Tecan AC Extraction Plate simplifies the sample preparation step in such a way that no centrifugation, filtration or solvent evaporation steps are needed. In fact the sample preparation procedure is reduced to simple pipetting and shaking. These pipetting and shaking steps were automated on a Tecan Freedom EVO liquid handling system. 

In this application note an approach is described which overcomes the difficulties described above.