This method achieves highly reproducible recoveries for 1,25 diOH Vitamins D2 and D3. A calibration range between 5 and 500 pg/mL is demonstrated using a starting volume of 0.25 mL of serum. Sensitivity is maximized through the use of a simple PTAD derivatization and formation of a methylamine complex.
The method was automated on the Biotage® Extrahera™, using ISOLUTE SLE+ 400 μL Supported Liquid Extraction plates. Full details of system setting are included in the application note.
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Application Video: Extraction Steroids from Serum with ISOLUTE® SLE+