Volumetric absorptive microsampling as an alternative sampling strategy for the determination of paracetamol in blood and cerebrospinal fluid
In the field of bioanalysis, dried matrix spot sampling is increasingly receiving interest, as this alternative sampling strategy offers many potential benefits over traditional sampling, including matrix volume-sparing properties. By using a microsampling strategy, e.g., volumetric absorptive microsampling (VAMS), the number of samples that can be collected from a patient can be increased, as a result of the limited sample volume that is required per sample. To date, no VAMS-based methods have been developed for the quantification of analytes in cerebrospinal fluid (CSF). The objective of this study was to develop and validate two LC-MS/MS methods for the quantification of paracetamol in dried blood and dried CSF, with both matrices sampled using VAMS. Both methods were fully validated based on internationally accepted guidelines. Paracetamol was chromatographically separated from its glucuronide and sulfate metabolites and no carry-over or unacceptable interferences were detected. The total precision (%RSD) was below 15% for all QC levels and accuracy (%bias) was below 7% (17% for the LLOQ of aqueous VAMS). The influence of the hematocrit on the recovery of blood VAMS samples appeared to be limited within the hematocrit range of 0.21 to 0.62. The blood VAMS samples were stable for 1 week if stored at 50 °C, and for at least 8 months when stored between − 80 °C and room temperature. The aqueous VAMS samples were stable for at least 9 months when stored between − 80 and 4 °C, and for 1 month when stored at room temperature. Application of the methods on external quality control material and analysis of patient samples demonstrated the validity and utility of the methods and provided a proof of concept for the analysis of CSF microsamples obtained via VAMS devices.
KeywordsVolumetric absorptive microsampling Liquid chromatography-tandem mass spectrometry Alternative sampling strategies Cerebrospinal fluid Paracetamol
Analysis of variance
Dried matrix spot
European Medicines Agency
External quality control
US Food and Drug Administration
Lower limit of quantification
Multiple reaction monitoring
Tandem mass spectrometry
Relative standard deviation
Upper limit of quantification
Volumetric absorptive microsampling
The authors wish to acknowledge Prof. Veronique Stove and her team for assistance with blood collection and hematocrit measurements and all volunteers who participated in the study.
This study was supported by the Research Foundation – Flanders (G0E010916N).
Compliance with ethical standards
Approval for the clinical proof-of-concept part of this study was provided by the Ethics Committee of Ghent University Hospital (B670201629325). Informed consent has been obtained from the participants involved or their legal representatives.
Conflict of interest
The authors declare that they have no conflict of interest.
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