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Hydrophilic Interaction Liquid chromatography/positive Ion Electrospray Ionization Mass Spectrometry Method for the Quantification of Alprazolam and α-hydroxy-alprazolam in Human Plasma

Hydrophilic Interaction Liquid chromatography/positive Ion Electrospray Ionization Mass Spectrometry Method for the Quantification of Alprazolam and α-hydroxy-alprazolam in Human Plasma

Eleni Kalogria, Constantinos Pistos, Irene Panderi

J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Dec 30;942-943:158-64.

PMID: 24269910

Abstract:

A hydrophilic interaction liquid chromatography/positive ion electrospray-mass spectrometry (HILIC-ESI/MS) has been developed and fully validated for the quantification of alprazolam and its main metabolite, α-hydroxy-alprazolam, in human plasma. The assay is based on 50μL plasma samples, following liquid-liquid extraction. All analytes and the internal standard (tiamulin) were separated by hydrophilic interaction liquid chromatography using an X-Bridge-HILIC analytical column (150.0mm×2.1mm i.d., particle size 3.5μm) under isoscratic elution. The mobile phase was composed of a 7% 10mM ammonium formate water solution in acetonitrile and pumped at a flow rate of 0.20mLmin(-1). Running in positive electrospray ionization and selected ion monitoring (SIM) the mass spectrometer was set to analyze the protonated molecules [M+H](+) at m/z 309, 325 and 494 for alprazolam, α-hydroxy-alprazolam and tiamulin (ISTD) respectively. The assay was linear over the concentration range of 2.5-250ngmL(-1) for alprazolam and 2.5-50ngmL(-1) for α-hydroxy alprazolam. Intermediate precision was less than 4.1% over the tested concentration ranges. The method is the first reported application of HILIC in the analysis benzodiazepines in human plasma. With a small sample size (50μL human plasma) and a run time less than 10.0min for each sample the method can be used to support a wide range of clinical studies concerning alprazolam quantification.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP37115438-A	alpha-Hydroxyalprazolam solution		37115-43-8	Price

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