Esmolol is a very short-acting beta-blocker commonly used in critically ill patients. To study the pharmacokinetics of esmolol in pediatric patients, a liquid chromatography-electrospray mass spectrometry (LC-MS) method to quantitate esmolol concentration in plasma was developed. Following methylene chloride extraction from 200 microl aliquots of plasma containing internal standard and reconstitution in 0.05% formic acid, 10 microl are injected onto column, eluted using a methanol/formic acid gradient over 15 min, and monitored with selected ion recording at 296.2 and 282.2. The assay is linear between 2 and 1000 ng/ml. The intra-day and inter-day coefficients of variation are less than 8 and 10%, respectively.