The maternal serum metabolome by multisegment injection-capillary electrophoresis-mass spectrometry: a high-throughput platform and standardized data workflow for large-scale epidemiological studies

Nat Protoc. 2021 Apr;16(4):1966-1994. doi: 10.1038/s41596-020-00475-0. Epub 2021 Mar 5.


A standardized data workflow is described for large-scale serum metabolomic studies using multisegment injection-capillary electrophoresis-mass spectrometry. Multiplexed separations increase throughput (<4 min/sample) for quantitative determination of 66 polar/ionic metabolites in serum filtrates consistently detected (coefficient of variance (CV) <30%) with high frequency (>75%) from a multi-ethnic cohort of pregnant women (n = 1,004). We outline a validated protocol implemented in four batches over a 7-month period that includes details on preventive maintenance, sample workup, data preprocessing and metabolite authentication. We achieve stringent quality control (QC) and robust batch correction of long-term signal drift with good mutual agreement for a wide range of metabolites, including serum glucose as compared to a clinical chemistry analyzer (mean bias = 11%, n = 668). Control charts for a recovery standard (mean CV = 12%, n = 2,412) and serum metabolites in QC samples (median CV = 13%, n = 202) demonstrate acceptable intermediate precision with a median intraclass coefficient of 0.87. We also report reference intervals for 53 serum metabolites from a diverse population of women in their second trimester of pregnancy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Glucose / metabolism
  • Calibration
  • Canada
  • Cohort Studies
  • Electrophoresis, Capillary / methods*
  • Electrophoresis, Capillary / standards*
  • Epidemiologic Studies*
  • Fasting / blood
  • Female
  • High-Throughput Screening Assays / methods*
  • Humans
  • Ions
  • Mass Spectrometry
  • Metabolome*
  • Metabolomics
  • Pregnancy
  • Principal Component Analysis
  • Quality Control
  • Reference Standards
  • Reference Values
  • Serum / metabolism*


  • Blood Glucose
  • Ions

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