, Volume 10, Issue 2, pp 312–323

Validation of a dual LC–HRMS platform for clinical metabolic diagnosis in serum, bridging quantitative analysis and untargeted metabolomics

  • Ilya Gertsman
  • Jon A. Gangoiti
  • Bruce A. Barshop
Original Article

DOI: 10.1007/s11306-013-0582-1

Cite this article as:
Gertsman, I., Gangoiti, J.A. & Barshop, B.A. Metabolomics (2014) 10: 312. doi:10.1007/s11306-013-0582-1


Mass spectrometry-based metabolomics is a rapidly growing field in both research and diagnosis. Generally, the methodologies and types of instruments used for clinical and other absolute quantification experiments are different from those used for biomarkers discovery and untargeted analysis, as the former requires optimal sensitivity and dynamic range, while the latter requires high resolution and high mass accuracy. We used a Q-TOF mass spectrometer with two different types of pentafluorophenyl (PFP) stationary phases, employing both positive and negative ionization, to develop and validate a hybrid quantification and discovery platform using LC–HRMS. This dual-PFP LC–MS platform quantifies over 50 clinically relevant metabolites in serum (using both MS and MS/MS acquisitions) while simultaneously collecting high resolution and high mass accuracy full scans to monitor all other co-eluting non-targeted analytes. We demonstrate that the linearity, accuracy, and precision results for the quantification of a number of metabolites, including amino acids, organic acids, acylcarnitines and purines/pyrimidines, meets or exceeds normal bioanalytical standards over their respective physiological ranges. The chromatography resolved highly polar as well as hydrophobic analytes under reverse-phase conditions, enabling analysis of a wide range of chemicals, necessary for untargeted metabolomics experiments. Though previous LC–HRMS methods have demonstrated quantification capabilities for various drug and small molecule compounds, the present study provides an HRMS quant/qual platform tailored to metabolic disease; and covers a multitude of different metabolites including compounds normally quantified by a combination of separate instrumentation.


Untargeted metabolomics Targeted metabolomics Bioanalytical validation Mass spectrometry Q-TOF LC–HRMS Comprehensive metabolite profiling 



Clinical laboratory improvement act


College for American pathologists


Good laboratory practices


Hepatocellular carcinoma


Cirrhotic patient


Not determined


Time of flight


(Lower) limit of quantification


Quality control


Coefficient of variation


High resolution mass spectrometry


Maple syrup urine disease


Medium chain acyl-CoA dehydrogenase deficiency



Supplementary material

11306_2013_582_MOESM1_ESM.docx (120 kb)
Supplementary material 1 (DOCX 119 kb)
11306_2013_582_MOESM2_ESM.jpg (92 kb)
Supplementary material 2 (JPEG 92 kb)
11306_2013_582_MOESM3_ESM.pdf (2.9 mb)
Supplementary material 3 (PDF 3002 kb)

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Ilya Gertsman
    • 1
  • Jon A. Gangoiti
    • 1
  • Bruce A. Barshop
    • 1
  1. 1.Biochemical Genetics LabUniversity of California, San DiegoLa JollaUSA

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