High-performance liquid chromatography-atmospheric pressure chemical ionisation-mass spectrometry determination of zaleplon in human plasma

A sensitive and specific liquid chromatography-atmospheric pressure chemical ionisation-mass spectrometry (LC-APCI-MS) method has been developed and validated for the identification and quantification of zaleplon in human plasma using estazolam as an internal standard (IS). After the addition of estazolam and 2.0 M sodium hydroxide solution. plasma samples were extracted with ethyl acetate and then the organic layer was evaporated to dryness. The reconstituted solution of the residue was injected onto a prepacked Shim-pack VP-ODS C-18 (250 mm x 2.0 mm i.d.) column and chromatographed with a mobile phase comprised of methanol: water (70:30) at a flow-rate of 0.2 ml/min. Detection was performed on a single quadrupole mass spectrometer by selected ion monitoring (SIM) mode via atmospheric pressure chemical ionization (APCI) source. The mean standard curve was linear (r = 0.9991) over the concentration range of 0.2-100 ng/ml and had good back-calculated accuracy and precision. The intra-day and inter-day precisions were within 10% relative standard deviation and accuracy ranged from 85% to 115%. The limit of detection was 0.1 ng/ml. The validated LC-APCI-MS method has been used successfully to study zaleplon pharmacokinetic, bioavailability and bioequivalence in 18 adult volunteers. (c) 2005 Elsevier B.V. All rights reserved.