Comparing derivatization reagents for quantitative LC–MS/MS analysis of a variety of vitamin D metabolites

作者：Αναστασία Αλεξανδρίδου, Pascal Schorr, Dietrich A. Volmer · 发表于：Analytical and Bioanalytical Chemistry · 年份：2023 · DOI：10.1007/s00216-023-04753-0 · 被引用次数：28 · 研究领域：Vitamin D Research Studies、Tryptophan and brain disorders、Effects and risks of endocrine disrupting chemicals

Abstract The present study systematically compares the sensitivity and selectivity of the analysis of multiple vitamin D metabolites after chemical derivatization using different reagents for liquid chromatography-tandem mass spectrometry (LC–MS/MS). Generally, chemical derivatization is applied to vitamin D metabolites to increase the ionization efficiency, which is particularly important for very low abundant metabolites. Derivatization can also improve the selectivity of the LC separation. A wide variety of derivatization reagents has been reported in recent years, but information on their relative performance and applicability to different vitamin D metabolites is, unfortunately, not available in the literature. To fill this gap, we investigated vitamin D 3 , 3β-25-hydroxyvitamin D 3 (3β-25(OH)D 3 ), 3α-25-hydroxyvitamin D 3 (3α-25(OH)D 3 ), 1,25-dihydroxyvitamin D 3 (1,25(OH) 2 D 3 ), and 24,25-dihydroxyvitamin D 3 (24,25(OH) 2 D 3 ) and compared response factors and selectivity after derivatizing with several important reagents, including four dienophile reagents (4-phenyl-1,2,4-triazoline-3,5-dione (PTAD), 4-[2-(6,7-dimethoxy-4-methyl-3-oxo-3,4-dihydroquinoxalinyl)ethyl]-1,2,4-triazoline-3,5-dione (DMEQ-TAD), Amplifex, 2-nitrosopyridine (PyrNO)) as well as two reagents targeting hydroxyl groups: isonicotinoyl chloride (INC) and 2-fluoro-1-methylpyridinium-p-toluenesulfonate (FMP-TS). In addition, a combination of dienophiles and hydroxyl group reagents was examined. Fo...