Beyond the limit of assignment of metabolites using minimal serum samples and 1H NMR spectroscopy with cross-validation by mass spectrometry

Ashish Gupta, Deepak Kumar

Research output: Contribution to journalArticle

Abstract

Identification of NMR-based metabolic indexes is limited by the deleterious effects of copious proteins and lipoproteins in the serum that accentuate the need for advance and high-throughput method. We tried to explore the use of a novel filtration (2KDa molecular weight cut-off) approach to remove the proteins from serum following use of less sample volume (only 150 μL of filtered serum), combining an array of 1D/2D NMR experiments (at 800 MHz spectrometer), spiking experiments with standard compounds, and validated by mass spectrometry. This novel method enabled the identification of a large number (n = 73) of metabolites and their percentage of abundance in the present study cohort. Mass spectrometry further validates and confirms the presence of all these 73 metabolites using same filtered serum. This study reveals seven new metabolites (citrulline, inosine, taurine, trimethyl amine, methylmalonate, uracil, methanol) in filtered serum using 1D/2D NMR spectroscopy that were not observed in earlier available literature using protein precipitation approach. This novel method delineates volatile metabolites, nitrogenous bases and nucleosides that may provide a milestone for the identification of inborn error of metabolism, pathogenicity at molecular level, disease identification and prognosis, and forensic studies using minimal volume of filtered serum samples and NMR spectroscopy.

Original languageEnglish (US)
Pages (from-to)356-364
Number of pages9
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume151
DOIs
StatePublished - Mar 20 2018
Externally publishedYes

Fingerprint

Metabolites
Nuclear magnetic resonance spectroscopy
Mass spectrometry
Mass Spectrometry
Magnetic Resonance Spectroscopy
Serum
Nuclear magnetic resonance
Inosine
Citrulline
Uracil
Taurine
Inborn Errors Metabolism
Nucleosides
Metabolism
Lipoproteins
Amines
Methanol
Spectrometers
Blood Proteins
Proteins

Keywords

  • Filtered serum
  • Mass spectrometer
  • Metabolic profiling
  • Metabolomics
  • NMR spectroscopy

ASJC Scopus subject areas

  • Analytical Chemistry
  • Pharmaceutical Science
  • Drug Discovery
  • Spectroscopy
  • Clinical Biochemistry

Cite this

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