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Simultaneous determination of vitamin D metabolites 25(OH)D~3~ and 1α,25(OH)~2~D~3~ in human plasma using liquid chromatography tandem mass spectrometry by Shan Xu; Rui Ni; Lihong Lv; Rui Chen; Yao Chen; Fengjiao Huang; Zhiru Xu is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Background: Although measurement of 25(OH)D 3 is a routine analytical method to determine plasma vitamin D status, 1α,25(OH) 2 D 3 is the biologically active form. Hence, simultaneous measurement of 25(OH)D 3 and 1α,25 (OH) 2 D 3 could provide better insight into vitamin D status and pharmacokinetics. However, 1α,25(OH) 2 D 3 has a low plasma concentration, making its quantification challenging for most analytical techniques. Here, we demonstrate use of liquid chromatography tandem mass spectrometry (LC-MSMS) for the development of a simple and rapid method for the simultaneous quantification of 25(OH)D 3 and 1α,25(OH) 2 D 3 . Methods: Samples were purified from 250 μL human plasma. Chromatography was performed on an analytical column, under gradient conditions using a mobile phase consisting of methanol-lithium acetate. The mass detector was operated in positive multiple reaction monitoring mode. The established method was validated according to the guidance issued by ICH and FDA. Furthermore, a clinical study was performed using this method to detect the plasma concentrations of 1α,25(OH) 2 D 3 after oral administration of calcitriol. ## Results and conclusion: The method was acc

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Author

Shan Xu; Rui Ni; Lihong Lv; Rui Chen; Yao Chen; Fengjiao Huang; Zhiru Xu

Publisher

Elsevier BV

Published

2022

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)