Abstract
Objectives
To update traditional “wet” matrices to dried blood spot (DBS) sampling, based on the liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) technique, and develop a method for simultaneous analyzing caffeine and its three primary metabolites (theobromine, paraxanthine, and theophylline), supporting routine therapeutic drug monitoring (TDM) for preterm infants.
Methods
DBS samples were prepared by a two-step quantitative sampling method, i.e., volumetric sampling of a quantitative 10 μL volume of peripheral blood and an 8 mm diameter whole punch extraction by a methanol/water (80/20, v/v) mixture containing 125 mM formic acid. Four paired stable isotope labeled internal standards and a collision energy defect strategy were applied for the method optimization. The method was fully validated following international guidelines and industrial recommendations on DBS analysis. Cross validation with previously developed plasma method was also proceeded. The validated method was then implemented on the TDM for preterm infants.
Results
The two-step quantitative sampling strategy and a high recovery extraction method were developed and optimized. The method validation results were all within the acceptable criteria. Satisfactory parallelism, concordance, and correlation were observed between DBS and plasma concentrations of the four analytes. The method was applied to provide routine TDM services to 20 preterm infants.
Conclusions
A versatile LC-MS/MS platform for simultaneous monitoring caffeine and its three primary metabolites was developed, fully validated, and successfully applied into the routine clinical TDM practices. Sampling method switching from “wet” matrices to “dry” DBS will facilitate and support the precision dosing of caffeine for preterm infants.
Funding source: Specially Appointed Medical Expert Project of the Jiangsu Commission of Health
Award Identifier / Grant number: 2019
Funding source: Scientific Research Support Foundation for Top Young Scholars at the Children’s Hospital of Nanjing Medical University
Award Identifier / Grant number: 2020
Funding source: Special Fund for Clinical Research of the Wu Jieping Medical Foundation
Award Identifier / Grant number: 320.6750.2020-04-07
Acknowledgments
The authors would like to thank the two anonymous reviewers for their constructive critical remarks and the editors of Clinical Chemistry and Laboratory Medicine for their kind suggestions and assistance along with all of the authors of the previous works that we referenced. These contributions helped significantly to refine and improve the content and the presentation of the paper.
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Research funding: This work was supported by the Specially Appointed Medical Expert Project of the Jiangsu Commission of Health (2019) and Special Fund for Clinical Research of the Wu Jieping Medical Foundation (320.6750.2020-04-07). This study was also supported by the Scientific Research Support Foundation for Top Young Scholars at the Children’s Hospital of Nanjing Medical University (2020).
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Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
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Competing interests: Authors state no conflict of interest.
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Informed consent: Informed consent was not required to participate in this study in accordance with the national legislation and the institutional requirements.
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Ethical approval: Research involving human subjects complied with all relevant national regulations, institutional policies and is in accordance with the tenets of the Helsinki Declaration (as revised in 2013), and has been approved by the authors’ Institutional Review Board (202012146-1).
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Supplementary Material
This article contains supplementary material (https://doi.org/10.1515/cclm-2023-0310).
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