A validated UPLC-MS/MS method for the quantitation of an unstable peptide, monocyte locomotion inhibitory factor (MLIF) in human plasma and its application to a pharmacokinetic study

Journal of Pharmaceutical and Biomedical Analysis
Xuemei LiuJi Jiang

Abstract

Monocyte locomotion inhibitory factor (MLIF, Met-Gln-Cys-Asn-Ser), a pentapeptide with anti-inflammatory activity, was developed for neural protection in acute ischemic stroke. Determination of MLIF in human plasma samples is of great importance for pharmacokinetic evaluation in clinical studies. A reliable and sensitive method based on ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) was established for the measurement of MLIF in human plasma. Instability of peptide in matrix was the primary challenge in method development, which was properly resolved by addition of acidification reagents like sulfuric acid. Samples were prepared by protein precipitation and then analyzed using a gradient chromatographic separation over an ACQUITY UPLC HSS T3 column. The mobile phase consisted of acetonitrile containing 0.2% formic acid and water containing 0.2% formic acid and gradient elution was performed at a flow rate of 0.4 mL/min. Detection was carried out on a Xevo TQ-S tandem mass spectrometer and positive electrospray ionization was employed in the multiple reaction monitoring (MRM) mode. This method was fully validated over the concentration range of 0.5-40 ng/mL with a lower limit of quanti...Continue Reading

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