Characterization of rapid extraction protocols for high‐throughput metabolomics. (25th July 2017)

• Record Type: Journal Article
• Authors: Gehrke, Sarah; Reisz, Julie A.; Nemkov, Travis; Hansen, Kirk C.; D'Alessandro, Angelo
• Is Part Of: Rapid communications in mass spectrometry. Volume 31:Number 17(2017)
• Page Start: 1445
• Page End: 1452
• Publication Date: 2017-07-25
• Subjects: Mass spectrometry -- Periodicals; 543.65
• DOI: 10.1002/rcm.7916
• Languages: English
• ISSNs: 0951-4198

Abstract

Abstract : Rationale: In the last five years, high‐throughput metabolomics has significantly advanced scientific research and holds the potential to promote strides in the fields of clinical metabolomics and personalized medicine. While innovations in the field of flow‐injection mass spectrometry and three‐minute metabolomics methods now allow investigators to process hundreds to thousands of samples per day, time‐sensitive clinical applications, particularly in the emergency department, are limited by a lack of rapid extraction methods. Methods: Here we characterized the efficacy of fast liquid–liquid extractions for characterization of hydrophilic compounds through ultra‐high‐pressure liquid chromatography/mass spectrometry. Internal stable‐isotope‐labeled standards were used to quantitatively characterize markers of energy and oxidative metabolism in human whole blood, plasma and red blood cells – three common matrices of clinical relevance. Results: For all the tested matrices, vortexing time (4–60 min) did not significantly affect extraction yields for the tested hydrophilic metabolites. Coefficients of variations <<20% for all tested compounds, except for the redox‐sensitive metabolite cystine (accumulating over time). Internal standards and second extractions confirmed recoveries >80% for all tested metabolites, except for basic amino acids and polyamines, which showed reproducible yields ranging from 50 to 75%. Global profiling and absolute quantitation of 24 metabolites revealed similarities between the plasma and red blood cell metabolomes. Conclusions: Rapid extraction (~4 min) of hydrophilic compounds is a viable and potentially automatable strategy to perform quantitative analysis of whole blood, plasma and red blood cells for research or clinical applications.