Determination of caffeine, paraxanthine, theophylline and theobromine in premature infants by HILIC-MS/MS

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, theophylline and theobromine in premature infants. Method: Protein precipitation of plasma samples (10 mu l) was carried out by treating with acetonitrile containing caffeine-C-13(3). The separation was carried out on an ACQUITY HPLC (R) 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 0.0600-60.0 mu g/ml for caffeine, 0.0250-7.50 mu g/ml for theobromine and 0.0150-4.50 mu g/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.

Automated summary

A HILIC-MS/MS method was developed and validated to determine the levels of caffeine and its major metabolites in the plasma of premature infants. This method showed good linearity and accuracy within the specified concentration ranges.