Vitamin D metabolites and analytical challenges

Naik, Megha; Kamath, Saritha U.; Shubhashree, U; Adiga, Satish Kumar; Patil, Ajeetkumar

doi:10.1039/d2ay01692c

Cited by 2 publications

(1 citation statement)

References 138 publications

(180 reference statements)

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“…As a promising alternative, LC-MS-based methods can reliably measure D2 and D3 metabolites because of their high selectivity with a molecular-level resolution. Nevertheless, achieving highly sensitive and accurate quantification of VitD metabolites while maintaining high throughput and robustness remains challenging for LC-MS-based methods 21 , 23 , 24 . Although derivatization is largely employed to enhance the ionization efficiency of VitD metabolites, enabling more sensitive LC-MS analysis 25 , the improvement in sensitivity may not be sufficient for measuring low-abundance VitD metabolites, especially in individuals with specific diseases where serum levels of these metabolites are anticipated to be lower than in healthy subjects 26 .…”

Section: Introductionmentioning

confidence: 99%

An ultra-sensitive and high-throughput trapping-micro-LC-MS method for quantification of circulating vitamin D metabolites and application in multiple sclerosis patients

Qu,

Zhang,

Weinstock-Guttman

et al. 2024

Sci Rep

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Quantitative analysis of the biologically-active metabolites of vitamin D (VitD), which are crucial in regulating various physiological and pathological processes, is important for clinical investigations. Liquid chromatography-tandem mass spectrometry (LC-MS) has been widely used for this purpose but existing LC-MS methods face challenges in achieving highly sensitive and accurate quantification of low-abundance VitD metabolites while maintaining high throughput and robustness. Here we developed a novel pipeline that combines a trapping-micro-LC-(T-µLC) with narrow-window-isolation selected-reaction monitoring MS(NWI-SRM) for ultra-sensitive, robust and high-throughput quantification of VitD metabolites in serum samples after derivatization. The selective-trapping and delivery approach efficiently removes matrix components, enabling high-capacity sample loading and enhancing sensitivity, throughput, and robustness. The NWI-SRM further improves the sensitivity by providing high selectivity. The lower limits of quantification (LOQs) achieved were markedly lower than any existing LC-MS methods: 1.0 pg/mL for 1,25(OH)2D3, 5.0 pg/mL for 24,25(OH)2D3, 30 pg/mL for both 25(OH)D2 and 25(OH)D3, all within a 9-min cycle. The method is applied to quantify VitD metabolites from 218 patients with multiple sclerosis. This study revealed negative correlations(r=− 0.44 to − 0.51) between the levels of 25(OH)D2 and all the three D3 metabolites in multiple sclerosis patients.

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