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Updated: Apr 26, 2026
Multi-step Preparation Technique to Recover Multiple Metabolite Compound Classes for In-depth and Informative Metabolomic Analysis
Published on: July 11, 2014
A novel method for the measurement of plasma metanephrines using online solid phase extraction-liquid chromatography
Joanne E Adaway1, Mirko Peitzsch2, Brian G Keevil3
1Department of Clinical Biochemistry, University Hospital South Manchester NHS Foundation Trust, Manchester, UK Manchester Academic Health Science Centre, University Hospital South Manchester, The University of Manchester, Manchester, UK Jo.adaway@uhsm.nhs.uk.
Background:
Measurement of plasma metanephrine, normetanephrine and 3-methoxytyramine is useful in the diagnosis of phaeochromocytomas, but many assays require a large volume of plasma due to poor assay sensitivity, and often require lengthy sample preparation. Our aim was to develop a method for measurement of plasma metanephrines using a small sample volume with minimal hands-on preparation.
Methods:
Samples were deproteinised using 10 K spin filters prior to online solid phase extraction using a Waters Acquity UPLC Online SPE Manager (Waters, Manchester, UK) coupled to a Waters Xevo TQ-S mass spectrometer (Waters, Manchester, UK). The assay was validated and results compared to a previously published method.
Results:
We achieved a limit of quantification of 37.5 pmol/L for metanephrine and 3-methoxytyramine and 75 pmol/L for normetanephrine using only 150 µL of sample. The assay was linear up to 30,000 pmol/L for all analytes and in a method comparison study results showed good agreement with a previously published LC-MS/MS assay.
Conclusions:
We have developed a simple method for measurement of plasma metanephrine, normetanephrine and 3-methoxytyramine using only 150 µL of sample. There is minimal hands-on sample preparation required and the assay is suitable for routine use in a clinical laboratory.
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