Ion-pair ultra-high performance liquid chromatographic analysis of monoamines: peak-splitting at high flow rates
Jolien Van Schoors1, Hendrik-Jan Brouwer, Katrien Maes
1Department of Pharmaceutical Chemistry and Drug Analysis, Center for Neurosciences, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium.
Abstract:
The use of ion-pair ultra-high performance liquid chromatography (UHPLC) coupled with electrochemical detection (ECD) is of great interest for the fast and sensitive determination of the monoamine neurotransmitters dopamine, noradrenaline and serotonin in microdialysis samples. However, when applying high flow rates in ion-pair UHPLC, other peaks than the initial compound peaks appear on the chromatogram. This peak-splitting phenomenon is caused by disturbed ion-pair retention mechanisms. The influence of several chromatographic parameters is investigated. Peak-splitting is delayed to higher flow rates when increasing the concentration of ion-pair reagent or buffering agent in the mobile phase, when decreasing the percentage of organic modifier in the mobile phase, when applying a stationary phase with a smaller amount of packing material or when increasing the separation temperature. One or a combination of these conditions can be applied to analyze the monoamine neurotransmitters using ion-pair UHPLC-ECD at high flow rates.
More Related Videos
09:34Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
Published on: September 20, 2016
10:17High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
Published on: April 23, 2019
Related Concept Videos
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
High-Performance Liquid Chromatography: Elution Process
High-Performance Liquid Chromatography: Types of Detectors
High-Performance Liquid Chromatography: Instrumentation
Mass Spectrometry: Amine Fragmentation
In amines, the number of nitrogen atoms affects the mass of the molecular ion, which is described by the nitrogen rule of mass spectrometry. This rule states that a compound containing...
Mass Spectrometry: Complex Analysis
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
