Abstract
Aim: Caffeine is a central nervous system stimulant, used to treat apnea of prematurity. A hydrophilic interaction chromatography-tandem mass spectrometry(HILIC-MS/MS) approach was developed to detect caffeine, paraxanthine, theophyllineand theobromine in premature infants. Method: Protein precipitation of plasma samples (10μl) was carried out by treating with acetonitrile containing caffeine-13C3. The separation was carried out on an ACQUITY HPLC® BEH HILIC column. Caffeine and its metabolites were quantified by multiple reaction monitoring modes with positive electrospray ionization. Results: The established method had a good linear relationship in the range of 0.0600-60.0μg/ml for caffeine, 0.0250-7.50μg/ml for theobromine and0.0150-4.50μg/ml for paraxanthine and theophylline. Conclusion: A HILIC-MS/MS method was developed and validated to determine caffeine and its major metabolites in plasma of premature infants.
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