DBS-LC-MS/MS assay for caffeine: validation and neonatal application.

Area of Science:

• Pharmacology
• Analytical Chemistry
• Neonatal Medicine

Background:

• Therapeutic drug monitoring (TDM) of caffeine is crucial for preterm infants.
• Dried blood spot (DBS) microsampling offers advantages for infant sampling.
• Challenges exist in applying DBS for TDM, necessitating method validation.

Purpose of the Study:

• To describe and validate a DBS-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for caffeine analysis.
• To assess the suitability of DBS microsampling for caffeine TDM in infants.
• To investigate potential limitations, such as hematocrit dependence, in DBS-based caffeine measurements.

Main Methods:

• Development and validation of a DBS-LC-MS/MS assay for caffeine.
• Evaluation of method performance characteristics, including accuracy and precision.
• Analysis of 96 paired infant plasma and DBS clinical samples.

Main Results:

• The validated DBS-LC-MS/MS method demonstrated hematocrit dependence.
• Caffeine concentrations measured in DBS samples were significantly lower than in concurrently collected plasma samples.
• The observed differences in caffeine levels between DBS and plasma were independent of hematocrit variations.

Conclusions:

• Extensive validation with real-life clinical samples is essential for DBS-based TDM methods.
• DBS-LC-MS/MS is a viable alternative to traditional methods for caffeine TDM and pharmacokinetic studies in preterm infants.
• The method's accuracy and reliability in diverse infant populations require further investigation.