Serum Vitamin D Metabolites by HPLC-MS/MS Combined with Differential Ion Mobility Spectrometry: Aspects of Sample Preparation Without Derivatization

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2023 May 13

PMID 37175816

Authors

Liliia Usoltseva

Vitaliy Ioutsi

Yuriy Panov

Mariya Antsupova

Liudmila Rozhinskaya

Galina Melnichenko

Natalia Mokrysheva

Affiliations

Soon will be listed here.

Abstract

In current clinical practice, a thorough understanding of vitamin D metabolism is in high demand both for patients with various diseases and for healthy individuals. Analytical techniques that provide simultaneous measurement of multiple metabolites are preferred. Herein, the development of an HPLC-DMS-MS/MS method for the quantitation of vitamin D compounds (25(OH)D, 25(OH)D, 1,25(OH)D, 3-epi-25(OH)D, 24,25(OH)D, and D) in serum is described. The selected sample preparation procedure based on the combination of liquid-liquid and solid-phase extraction, which excluded a lengthy derivatization step, was compared with other common approaches. Sensitivity was increased through the implementation of differential ion mobility separation. The proposed assay allowed us to determine the low abundant 1,25(OH)D with the detection limit of 10 pg/mL. The validation study showed good linearity (r > 0.99), a wide analytical range (2.5-75 ng/mL for 25(OH)D), and acceptable precision (<7%) for all metabolites. The recovery ranged from 71% to 93% and the matrix effect from 0.80 to 0.95 depending on the metabolite; accuracy determination was performed using DEQAS controls.

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