Analysis of 25 underivatized amino acids in human plasma using ion-pairing reversed-phase liquid chromatography/time-of-flight mass spectrometry

Amino acids in biological fluids have previously been shown to be detectable using liquid chromatography/electrospray ionization mass spectrometry (LC/ESI‐MS) with perfluorinated acids as ion‐pairing agents. To date, these studies have used precursor mass, retention time and tandem mass spectrometry...

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Bibliographic Details
Published in:Rapid communications in mass spectrometry 2007-08, Vol.21 (16), p.2717-2726
Main Authors: Armstrong, Michael, Jonscher, Karen, Reisdorph, Nichole A.
Format: Article
Language:English
Subjects:
Amino Acids - blood
Amino Acids - chemistry
Blood Chemical Analysis - methods
Chromatography, High Pressure Liquid - methods
Chromatography, Ion Exchange - methods
Humans
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization - methods
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Summary:Amino acids in biological fluids have previously been shown to be detectable using liquid chromatography/electrospray ionization mass spectrometry (LC/ESI‐MS) with perfluorinated acids as ion‐pairing agents. To date, these studies have used precursor mass, retention time and tandem mass spectrometry (MS/MS) to identify and quantify amino acids. While this is a potentially powerful technique, we sought to adapt the method to time‐of‐flight (TOF)MS. A new application of a recently described liquid chromatographic separation method was coupled with TOFMS to employ accurate mass for qualitative identification; resulting in additional qualitative data not available with standard single quadrupole data. In the current study, we evaluated 25 physiological amino acids and one dipeptide that are routinely quantified in human plasma. Accuracy and precision of the method was evaluated by spiking human plasma with a mix of the 25 amino acids; in addition, the inclusion of a cation‐exchange cleanup step was evaluated. The calibration curves were linear over a range from 1.56 to 400 µM. The dynamic range was found to be within physiological levels for all amino acids analyzed. Accuracy and precision for most of the amino acids was between 80–120% spike recovery and
ISSN:0951-4198
1097-0231
DOI:10.1002/rcm.3124