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Spotlight on Endocannabinoids in Healthy Individuals Using Volumetric Absorptive Microsampling Combined with LC-MS/MS

Lounes Haroune1,2, Sabrina Saibi1,2, Jean-Philippe Bellenger3

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Volumetric absorptive microsampling (VAMS) offers a viable method for analyzing endocannabinoids (eCBs) in blood. However, sample matrix and drying time significantly impact eCB levels, necessitating standardized protocols for accurate clinical research.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Pharmacology

Background:

  • Endocannabinoids (eCBs) are vital lipid signaling molecules regulating numerous physiological processes.
  • Accurate quantification of eCBs is essential for understanding their role in health and disease.
  • Current analytical methods may face challenges with sample stability and matrix effects.

Purpose of the Study:

  • To evaluate the applicability of volumetric absorptive microsampling (VAMS) for analyzing endocannabinoids (eCBs) in blood.
  • To develop and validate a sensitive LC-MS/MS method for quantifying key eCBs and related lipid mediators.
  • To compare eCB levels in whole blood versus plasma samples and assess VAMS performance under various conditions.

Main Methods:

  • Development and validation of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for eCBs and eCB-like mediators.
  • Evaluation of VAMS performance, including stability testing at different temperatures, humidity, and hematocrit levels.
  • Systematic comparison of eCB concentrations in whole blood and plasma samples.

Main Results:

  • The LC-MS/MS method demonstrated high accuracy (72.5–98.9%) and precision (<8%) with minimal matrix effects (-15.0% to +9.0%).
  • Most eCBs were stable under various storage conditions, but drying time affected 2-AG and 2-OG levels, suggesting potential ex vivo biosynthesis.
  • Significant differences in eCB levels were observed between whole blood and plasma, with whole blood showing substantially higher 2-AG concentrations.

Conclusions:

  • VAMS is a promising technique for eCB analysis, but careful consideration of sample matrix and drying protocols is crucial.
  • Substantial distribution differences of eCBs between whole blood and plasma highlight the importance of cellular components.
  • Standardized sampling and analytical methods are imperative for reliable and reproducible eCB measurements in clinical research.