Propranolol is now a preferred treatment for infantile
hemangioma. However, there are no published papers on the metabolism and concentrations of
propranolol in the plasma of infants with
hemangioma. In the present study, a sensitive, simple and reliable method was developed and validated for the simultaneous quantification of
propranolol and its metabolites
4-hydroxypropranolol (M1) and
N-desisopropylpropranolol (M2) in infants' plasma for the first time by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A volume of 100 μL plasma was prepared by one-step
protein precipitation with
acetonitrile (300 μL), followed by its separation on an Hypersil
GOLD C18 column maintained at 40 °C with gradient mobile phase consisting of 0.1%
formic acid aqueous
solution and
acetonitrile at a flow rate of 0.3 mL min-1. The quantification was performed via multiple reaction monitoring (MRM) by a triple quadrupole mass spectrometer under positive electrospray ionization (ESI) mode.
Bisoprolol was chosen as the internal standard. The method was validated to demonstrate its selectivity, linearity, accuracy, precision, recovery, matrix effect and stability. The matrix-matched calibration curves for
propranolol ranging from 1 to 500 ng mL-1, for M1 ranging from 0.2 to 100 ng mL-1 and for M2 ranging from 0.2 to 100 ng mL-1 were all linear, with correlation coefficients calculated using weighted (1/x 2) least square linear regression analysis. The lower limits of quantification (LLOQs) were 1 ng mL-1, 0.2 ng mL-1 and 0.2 ng mL-1 for
propranolol, M1 and M2, respectively. The intra-day and inter-day precisions were less than 7.1% and relative errors were all less than 9.8%. This validated method was successfully applied to quantify the concentrations of
propranolol and its metabolites
4-hydroxypropranolol (M1) and
N-desisopropylpropranolol (M2) in the plasma of infants with
hemangioma after
oral administration of different doses of
propranolol for the first time.