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Quantifying precision loss in targeted metabolomics based on mass spectrometry and non-matching internal standards

Quantifying precision loss in targeted metabolomics based on mass spectrometry and non-matching internal standards
Quantifying precision loss in targeted metabolomics based on mass spectrometry and non-matching internal standards

In mass spectrometry, reliable quantification requires correction for variations in ionization efficiency between samples. The preferred method is the addition of a stable isotope-labeled internal standard (SIL-IS). In targeted metabolomics, a dedicated SIL-IS for each metabolite of interest may not always be realized due to high cost or limited availability. We recently completed the analysis of more than 70 biomarkers, each with a matching SIL-IS, across four mass spectrometry-based platforms (one GC-MS/MS and three LC-MS/MS). Using data from calibrator and quality control samples added to 60 96-well trays (analytical runs), we calculated analytical precision (CV) retrospectively. The use of integrated peak areas for all metabolites and internal standards allowed us to calculate precision for all matching analyte (A)/SIL-IS (IS) pairs as well as for all nonmatching A/IS pairs within each platform (total n = 1442). The median between-run precision for matching A/IS across the four platforms was 2.7-5.9%. The median CV for nonmatching A/IS (corresponding to pairing analytes with a non-SIL-IS) was 2.9-10.7 percentage points higher. Across all platforms, CVs for nonmatching A/IS increased with increasing difference in retention time (Spearman's rho of 0.17-0.93). The CV difference for nonmatching vs matching A/IS was often, but not always, smaller when analytes and internal standards were close structural analogs.

0910-6340
7616-7624
Ulvik, Arve
ad30af8c-4e72-4d2f-948c-547ad499c52a
McCann, Adrian
c4e41867-1522-4563-b4c7-4cc98a5c4696
Midttun, Oivind
35a547e1-094a-4073-be2d-38b28a4f583a
Meyer, Klaus
1ca5d2ca-1812-4dee-a02e-edb827194eb3
Godfrey, Keith
0931701e-fe2c-44b5-8f0d-ec5c7477a6fd
Ueland, Per M
43a3b904-232c-4c6d-9550-c8b3b55dd472
Ulvik, Arve
ad30af8c-4e72-4d2f-948c-547ad499c52a
McCann, Adrian
c4e41867-1522-4563-b4c7-4cc98a5c4696
Midttun, Oivind
35a547e1-094a-4073-be2d-38b28a4f583a
Meyer, Klaus
1ca5d2ca-1812-4dee-a02e-edb827194eb3
Godfrey, Keith
0931701e-fe2c-44b5-8f0d-ec5c7477a6fd
Ueland, Per M
43a3b904-232c-4c6d-9550-c8b3b55dd472

Ulvik, Arve, McCann, Adrian, Midttun, Oivind, Meyer, Klaus, Godfrey, Keith and Ueland, Per M (2021) Quantifying precision loss in targeted metabolomics based on mass spectrometry and non-matching internal standards. Analytical Sciences, 93 (21), 7616-7624. (doi:10.1021/acs.analchem.1c00119).

Record type: Article

Abstract

In mass spectrometry, reliable quantification requires correction for variations in ionization efficiency between samples. The preferred method is the addition of a stable isotope-labeled internal standard (SIL-IS). In targeted metabolomics, a dedicated SIL-IS for each metabolite of interest may not always be realized due to high cost or limited availability. We recently completed the analysis of more than 70 biomarkers, each with a matching SIL-IS, across four mass spectrometry-based platforms (one GC-MS/MS and three LC-MS/MS). Using data from calibrator and quality control samples added to 60 96-well trays (analytical runs), we calculated analytical precision (CV) retrospectively. The use of integrated peak areas for all metabolites and internal standards allowed us to calculate precision for all matching analyte (A)/SIL-IS (IS) pairs as well as for all nonmatching A/IS pairs within each platform (total n = 1442). The median between-run precision for matching A/IS across the four platforms was 2.7-5.9%. The median CV for nonmatching A/IS (corresponding to pairing analytes with a non-SIL-IS) was 2.9-10.7 percentage points higher. Across all platforms, CVs for nonmatching A/IS increased with increasing difference in retention time (Spearman's rho of 0.17-0.93). The CV difference for nonmatching vs matching A/IS was often, but not always, smaller when analytes and internal standards were close structural analogs.

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Accepted/In Press date: 28 April 2021
e-pub ahead of print date: 20 May 2021
Published date: 1 June 2021
Additional Information: Publisher Copyright: © 2021 American Chemical Society. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

Identifiers

Local EPrints ID: 448973
URI: http://eprints.soton.ac.uk/id/eprint/448973
ISSN: 0910-6340
PURE UUID: 0fbf3cfb-fff6-4a3d-b069-8f31abb9fd2c
ORCID for Keith Godfrey: ORCID iD orcid.org/0000-0002-4643-0618

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Date deposited: 12 May 2021 16:32
Last modified: 26 Nov 2021 02:35

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Contributors

Author: Arve Ulvik
Author: Adrian McCann
Author: Oivind Midttun
Author: Klaus Meyer
Author: Keith Godfrey ORCID iD
Author: Per M Ueland

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