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

• Published in: Nature protocols Vol. 16; no. 4; pp. 1966 - 1994
• Main Authors: Shanmuganathan, Meera, Kroezen, Zachary, Gill, Biban, Azab, Sandi, de Souza, Russell J., Teo, Koon K., Atkinson, Stephanie, Subbarao, Padmaja, Desai, Dipika, Anand, Sonia S., Britz-McKibbin, Philip
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.04.2021; Nature Publishing Group
• Subjects: 631/1647/320; 639/638/11/296; Analytical Chemistry; Biological Techniques; Biomedical and Life Sciences; Blood Glucose - metabolism; Calibration; Canada; Capillary electrophoresis; Cohort Studies; Computational Biology/Bioinformatics; Control charts; Electrophoresis; Electrophoresis, Capillary - methods; Electrophoresis, Capillary - standards; Epidemiologic methods; Epidemiologic Studies; Epidemiology; Fasting - blood; Female; High-Throughput Screening Assays - methods; Humans; Injection; Ions; Life Sciences; Mass Spectrometry; Mass spectroscopy; Mean; Medical examination; Metabolites; Metabolome; Metabolomics; Methods; Microarrays; Organic Chemistry; Pregnancy; Pregnant women; Preventive maintenance; Principal Component Analysis; Protocol; Quality Control; Reference Standards; Reference Values; Robust control; Scientific imaging; Serum - metabolism; Spectroscopy; Workflow; Canada
• ISSN: 1754-2189; 1750-2799
• DOI: 10.1038/s41596-020-00475-0

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. A standardized protocol and data workflow for high-throughput analysis of the maternal serum metabolome is outlined. It uses multisegment injection–capillary electrophoresis–mass spectrometry and is applied to a multi-ethnic cohort of pregnant women.