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Methylmalonic Acid: Evaluation of Sample Preparation and Simplicity of Method Implementation

Using Automated Sample Preparation Prior to LC-MS/MS Analysis

Introduction

The screening for elevated levels of methylmalonic acid (MMA) in serum is commonly used as a clinical diagnostic indicator of Cobalamin (Vitamin B12) deficiency in humans. MMA is commonly analyzed using LC-MS/MS with or without prior derivatization. This poster summarizes various sample preparation strategies for the extraction of MMA from serum without the necessity for derivatization, prior to LC-MS/MS analysis. A range of extraction techniques of varying complexity were evaluated: protein precipitation, phospholipid depletion, supported liquid extraction and solid phase extraction using both silica and polymer-based mixed-mode anion exchange chemistries.

Method performance was evaluated for evaporative effects, assay recovery, ion suppression, phospholipid removal and simplicity of transfer to an automated sample preparation platform.

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