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Ultra-Sensitive Method for Determining Vitamin D2 and D3 Metabolites in Serum

14 March 2016

A new application from Biotage Labs describes a protocol for the extraction of 1,25 di-OH Vitamin D2 and 1,25 di-OH Vitamin D3 metabolites from serum using supported liquid extraction prior to LC-MS/MS detection.


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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Download the application note below.