A Simultaneous Extraction/Derivatization Strategy for Quantitation of Vitamin D in Dried Blood Spots Using LC–MS/MS: Application to Biomarker Study in Subjects Tested for SARS-CoV-2

Vitamin D plays a critical role in bone development and maintenance, and in other physiological functions. The quantitation of endogenous levels of individual vitamin D and its metabolites is crucial for assessing several disease state conditions. With cases of severe acute respiratory syndrome coro...

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Veröffentlicht in:International journal of molecular sciences Jg. 24; H. 6; S. 5489
Hauptverfasser: Chhonker, Yashpal S., Ahmed, Nusrat, Johnston, Christine M., Barnabas, Ruanne V., Murry, Daryl J.
Format: Journal Article
Sprache:Englisch
Veröffentlicht: Switzerland MDPI AG 13.03.2023
MDPI
Schlagworte:
Alfacalcidol
Biomarkers
Blood
Calcifediol
Chromatography
Chromatography, Liquid - methods
Coronaviruses
COVID-19
Diagnosis
Disease
Formic acid
Health aspects
Humans
Infections
Ions
Medical research
Medicine, Experimental
Metabolites
Organic acids
Pandemics
Physiological aspects
Physiology
Plasma
Reproducibility of Results
Retention
SARS-CoV-2
Severe acute respiratory syndrome
Severe acute respiratory syndrome coronavirus 2
Tandem Mass Spectrometry - methods
Vitamin D
Vitamins
Massachusetts
biomarker
LC–MS/MS
SARS-CoV-2(+)
vitamin D
ISSN:1422-0067, 1661-6596, 1422-0067
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Zusammenfassung:Vitamin D plays a critical role in bone development and maintenance, and in other physiological functions. The quantitation of endogenous levels of individual vitamin D and its metabolites is crucial for assessing several disease state conditions. With cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) leading to the coronavirus disease 2019 (COVID-19) pandemic, there are several studies that have associated lower levels of serum vitamin D with severity of infection in COVID-19 patients. In this context, we have developed and validated a robust LC–MS/MS method for simultaneous quantitation of vitamin D and its metabolites in human dried blood spot (DBS) obtained from participants tested for COVID-19. The chromatographic separation for vitamin D and metabolites was performed using an ACE Excel C18 PFP column protected with a C18 guard column (Phenomenex, Torrance, CA, USA). The mobile phase consisted of formic acid in water (0.1% v/v) as mobile phase A and formic acid in methanol (0.1% v/v) as mobile phase B, operated at a flow rate of 0.5 mL/min. Analysis was performed utilizing the LC–MS/MS technique. The method was sensitive with a limit of quantification of 0.78 ng/mL for all analytes, and had a large dynamic range (200 ng/mL) with a total run time of 11 min. The inter- and intraday accuracy and precision values met the acceptance criteria per the US Food and Drug Administration guidelines. Blood concentrations of 25(OH)D3, vitamin D3, 25(OH)D2, and vitamin D2 over a range of 2–195.6, 0.5–121.5, 0.6–54.9, and 0.5–23.9 ng/mL, respectively, were quantified in 909 DBS samples. In summary, our developed LC−MS/MS method may be used for quantification of vitamin D and its metabolites in DBS, and may be applied to investigations of the emerging role of these compounds in various physiological processes.
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These authors contributed equally to this work.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms24065489