title: Sample Preparation Strategies for the Effective Quantitation of Hydrophilic Metabolites in Serum by Multi-Targeted HILIC-MS/MS.
abstract: The effect of endogenous interferences of serum in multi-targeted metabolite profiling HILIC-MS/MS analysis was investigated by studying different sample preparation procedures. A modified QuEChERS dispersive SPE protocol, a HybridSPE protocol, and a combination of liquid extraction with protein precipitation were compared to a simple protein precipitation. Evaluation of extraction efficiency and sample clean-up was performed for all methods. SPE sorbent materials tested were found to retain hydrophilic analytes together with endogenous interferences, thus additional elution steps were needed. Liquid extraction was not shown to minimise matrix effects. In general, it was observed that a balance should be reached in terms of recovery, efficient clean-up, and sample treatment time when a wide range of metabolites are analysed. A quick step for removing phospholipids prior to the determination of hydrophilic endogenous metabolites is required, however, based on the results from the applied methods, further studies are needed to achieve high recoveries for all metabolites.
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece. tsakeliz@hotmail.com.
authors.forename: Elisavet
authors.initials: E
authors.lastname: Tsakelidou
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece. cr_virgi@hotmail.com.
authors.forename: Christina
authors.initials: C
authors.lastname: Virgiliou
authors.affiliation: Laboratory of Forensic Medicine & Toxicology, Department of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece. lianavalianou@yahoo.gr.
authors.forename: Lemonia
authors.initials: L
authors.lastname: Valianou
authors.affiliation: Laboratory of Forensic Medicine & Toxicology, Department of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece. gkikae@auth.gr.
authors.forename: Helen G
authors.initials: HG
authors.lastname: Gika
authors.affiliation: Laboratory of Forensic Medicine & Toxicology, Department of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece. raikos@auth.gr.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Raikos
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece. gtheodor@chem.auth.gr.
authors.forename: Georgios
authors.initials: G
authors.lastname: Theodoridis
keywords: endogenous metabolites; sample preparation; serum; targeted metabolomics