Clinical validation of a DBS-based LC-MS/MS method for 25-hydroxyvitamin D: from lab sampling to home sampling
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Liesl Heughebaert
Abstract
Objectives
As (a lack of) vitamin D has been linked to a wide variety of chronic diseases, there is growing interest in generating robust epidemiological data. Consequently, the need for large-scale biological sample collection has increased. In this context, dried blood spot (DBS) microsampling offers a minimally invasive alternative to venous sampling. However, to allow interpretation of DBS-based 25-hydroxyvitamin D (25-(OH)D) results, the set-up of a clinical validation study is essential to assess the agreement with the reference matrix, plasma.
Methods
Venous plasma and whole blood, venous DBS (vDBS) and capillary DBS (cDBS) were collected from 44 healthy volunteers to evaluate the agreement between the different sample types for 25-(OH)D quantification. To transform cDBS-based results to plasma concentrations, a hematocrit (Hct)-dependent conversion factor was applied and evaluated using four different Hct determination approaches. All samples were analysed using previously described validated LC-MS/MS methods.
Results
No clinically relevant methodological (vDBS vs. whole blood) or sampling-site related (cDBS vs. vDBS) issues were observed. Following Hct-dependent conversion, good agreement between the cDBS-derived and actually measured plasma results was obtained, as 90 % of the results lay within 20 % of the plasma result, independent of the Hct approach used. Additionally, weighted Cohen’s kappa values of 0.83–0.85 were obtained across the different Hct approaches, indicating substantial to almost perfect agreement in vitamin D status classification.
Conclusions
Following Hct-dependent conversion, cDBS can be used as a reliable and practical alternative matrix to plasma for large-scale monitoring of vitamin D status in epidemiological and public health contexts.
Acknowledgments
The authors would like to acknowledge the team of prof. dr. Veronique Stove of the clinical laboratory from Ghent University Hospital for their help with the hematocrit measurements and the team of Heidi Lammens for their assistance with the venous blood draws. The authors also wish to thank all volunteers who participated in the study. Acquisition of the Waters Acquity UPLC® I-class plus coupled to a Waters XEVO TQ-XS was within the Ghent University MS-Small consortium, of which CS is part, and was made possible via financial support by the Ghent University Special Research Fund.
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Research ethics: Document ID: ONZ-2023-0284. Date grant: 2023-08-09.
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Informed consent: Informed consent was obtained from all individuals included in this study.
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Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission. Liesl Heughebaert: Writing – review and editing, Writing – original draft, Visualization, Validation, Investigation, Conceptualization. Rosalie Ghesquière: Writing – review and editing, Writing – original draft, Visualization, Validation, Investigation, Conceptualization. Christophe P. Stove: Writing – review and editing, Writing – original draft, Supervision, Project administration, Funding acquisition, Conceptualization.
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Use of Large Language Models, AI and Machine Learning Tools: During the preparation of this work the authors used ChatGPT in order to enhance the clarity and linguistic quality of the text. Following usage, the authors reviewed and edited the content as needed and take full responsibility for the content of the published article.
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Conflict of interest: The authors state no conflict of interest.
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Research funding: Liesl Heughebaert would like to thank the Special Research Fund (BOF) for granting her a PhD fellowship (01D05220). Rosalie Ghesquière would like to thank the Research Foundation Flanders (FWO) for granting her a PhD fellowship (1S88926N).
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Data availability: Data will be made available on request.
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