Extraction of monohydroxy and dihydroxy Vitamin D metabolites from serum

Extraction of 1,25 di-OH Vitamin D2, 1,25 di-OH Vitamin D3, 25 OH Vitamin D2 and 25 OH Vitamin D3 from serum Using ISOLUTE® SLE+ prior to LC-MS/MS Analysis

This application note describes a Supported Liquid Extraction (SLE) protocol for the extraction of both forms of 1,25 diOH Vitamin D metabolite and both forms of 25 OH metabolite in serum using ISOLUTE®SLE+ plates with LC-MS/MS detection.

Introduction

The method described in this application note achieves high recoveries of the 1,25 diOH and 25 OH metabolites of vitamins D2 and D3 from serum. The method is versatile enough to measure both the commonly measured 25 hydroxy metabolites and sensitive enough to also measure the better predicting 1,25 dihydroxy metabolites. The method involves SLE extraction followed by a simple PTAD derivatization.

ISOLUTE^® SLE+ products provide clean, rapid, robust and efficient extraction solutions for a wide range of analytes.