Quantitative analysis of phenylalanine, tyrosine, tryptophan and kynurenine in rat model for tauopathies by ultra-high performance liquid chromatography with fluorescence and mass spectrometry detection

Journal of Pharmaceutical and Biomedical Analysis
Jaroslav GalbaAndrej Kovac

Abstract

We developed and validated a simple and sensitive ultra-high performance liquid chromatography (UHPLC) method for the analysis of phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp) and kynurenine (Kyn) in rat plasma. Analytes were separated on Acquity UPLC HSS T3 column (2.1 mm×50 mm, 1.8 μm particle size) using a 4 min ammonium acetate (pH 5) gradient and detected by fluorescence and positive ESI mass spectrometry. Sample preparation involved dilution of plasma, deproteinization by trichloroacetic acid and centrifugation. The procedure was validated in compliance with the FDA guideline. The limits of quantification (LOQ) were 0.3 μM for Kyn and from 1.5 to 3 μM for Phe, Tyr, Trp. The method showed excellent linearity with regression coefficients higher than 0.99. The accuracy was within the range of 86-108%. The inter-day precision (n=5 days), expressed as % RSD, was in the range 1-13%. The benefit of using UHPLC is a short analysis period and thus, a very good sample throughput. Using this method, we analyzed plasma samples and detected significant changes of Kyn and Phe in transgenic rat model for tauopathies.

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Citations

Jun 28, 2016·Journal of Pharmaceutical and Biomedical Analysis·Rhishikesh ThakareYazen Alnouti
Jun 8, 2017·Journal of Separation Science·Ilona SadokMagdalena Maria Staniszewska
Aug 28, 2019·Biomedical Chromatography : BMC·Yan-Yan LiZheng-Chao Ji
Jun 6, 2018·Journal of Alzheimer's Disease : JAD·Petr NovakMichal Novak
Nov 9, 2018·Analytical Chemistry·Valentina D'AtriDavy Guillarme
Feb 8, 2020·Journal of the American Society for Mass Spectrometry·Burcu EserAylin Sepici Dinçel

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