Comparison of
Microflow and Analytical Flow Liquid
Chromatography Coupled to Mass Spectrometry Global Metabolomics Methods
Using a Urea Cycle Disorder Mouse Model
posted on 2021-11-29, 20:16authored bySarah Geller, Harvey Lieberman, Adam J. Belanger, Nelson S. Yew, Alla Kloss, Alexander R. Ivanov
Microscale-based
separations are increasingly being applied in
the field of metabolomics for the analysis of small-molecule metabolites.
These methods have the potential to provide improved sensitivity,
less solvent waste, and reduced sample-size requirements. Ion-pair
free microflow-based global metabolomics methods, which we recently
reported, were further compared to analytical flow ion-pairing reagent
containing methods using a sample set from a urea cycle disorder (UCD)
mouse model. Mouse urine and brain homogenate samples representing
healthy, diseased, and disease-treated animals were analyzed by both
methods. Data processing was performed using univariate and multivariate
techniques followed by analyte trend analysis. The microflow methods
performed comparably to the analytical flow ion-pairing methods with
the ability to separate the three sample groups when analyzed by partial
least-squares analysis. The number of detected metabolic features
present after each data processing step was similar between the microflow-based
methods and the ion-pairing methods in the negative ionization mode.
The observed analyte trend and coverage of known UCD biomarkers were
the same for both evaluated approaches. The 12.5-fold reduction in
sample injection volume required for the microflow-based separations
highlights the potential of this method to support studies with sample-size
limitations.