Development of nanoelectrospray high resolution isotope dilution mass spectrometry for targeted quantitative analysis of urinary metabolites: application to population profiling and clinical studies

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Title: Development of nanoelectrospray high resolution isotope dilution mass spectrometry for targeted quantitative analysis of urinary metabolites: application to population profiling and clinical studies
Authors: Chekmeneva, E
Correia, G
Denes, J
Gomez-Romero, M
Wijeyesekera, A
Perenyi, DR
Koot, Y
Boomsma, C
Want, EJ
Dixon, PH
Macklon, NS
Chan, Q
Takats, Z
Nicholson, JK
Holmes, E
Item Type: Journal Article
Abstract: An automated chip-based electrospray platform was used to develop a high-throughput nanoelectrospray high resolution mass spectrometry (nESI-HRMS) method for multiplexed parallel untargeted and targeted quantitative metabolic analysis of urine samples. The method was demonstrated to be suitable for metabolic analysis of large sample numbers and can be applied to large-scale epidemiological and stratified medicine studies. The method requires a small amount of sample (5 μL of injectable volume containing 250 nL of original sample), and the analysis time for each sample is three minutes per sample to acquire data in both negative and positive ion modes. Identification of metabolites was based on the high resolution accurate mass and tandem mass spectrometry using authentic standards. The method was validated for 8 targeted metabolites and was shown to be precise and accurate. The mean accuracy of individual measurements being 106% and the intra- and inter-day precision (expressed as relative standard deviations) were 9% and 14%, respectively. Selected metabolites were quantified by standard addition calibration using the stable isotope labelled internal standards in a pooled urine sample, to account for any matrix effect. The multiple point standard addition calibration curves yielded correlation coefficients greater than 0.99, and the linear dynamic range was more than three orders of magnitude. As a proof-of-concept the developed method was applied for targeted quantitative analysis of a set of 101 urine samples obtained from female participants with different pregnancy outcomes. In addition to the specifically targeted metabolites, several other metabolites were quantified relative to the internal standards. Based on the calculated concentrations, some metabolites showed significant differences according to different pregnancy outcomes. The acquired high resolution full-scan data were used for further untargeted fingerprinting and improved the differentiation of urine samples based on pregnancy outcome.
Issue Date: 28-May-2015
Date of Acceptance: 15-May-2015
ISSN: 1759-9679
Publisher: Royal Society of Chemistry
Start Page: 5122
End Page: 5133
Journal / Book Title: Analytical Methods
Volume: 7
Issue: 12
Copyright Statement: © 2015 The Royal Society of Chemistry.
Sponsor/Funder: National Institutes of Health
Medical Research Council (MRC)
Imperial College Healthcare NHS Trust- BRC Funding
Imperial College Healthcare NHS Trust- BRC Funding
Funder's Grant Number: 0600 370 D330 1362
RDB03 79560
RDB03 79560
Keywords: Science & Technology
Physical Sciences
Life Sciences & Biomedicine
Chemistry, Analytical
Food Science & Technology
0301 Analytical Chemistry
0399 Other Chemical Sciences
Publication Status: Published
Appears in Collections:Division of Surgery
Epidemiology, Public Health and Primary Care

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