Quantitative Analysis and Molecular Species Fingerprinting of Triacylglyceride Molecular Species Directly from Lipid Extracts of Biological Samples by Electrospray Ionization Tandem Mass Spectrometry
Abstract
Herein we describe a rapid, simple, and reliable method for the quantitative analysis and molecular species fingerprinting of triacylglycerides (TAG) directly from chloroform extracts of biological samples. Previous attempts at direct TAG quantitation by positive-ion electrospray ionization mass spectrometry (ESI/MS) were confounded by the presence of overlapping peaks from choline glycerophospholipids requiring chromatographic separation of lipid extracts prior to ESI/MS analyses. By exploiting the rapid loss of phosphocholine from choline glycerophospholipids, in conjunction with neutral-loss scanning for individual fatty acids, overlapping peaks in the ESI mass spectrum were deconvoluted generating a detailed molecular species fingerprint of individual TAG molecular species directly from chloroform extracts of biological samples. This method readily detects as little as 0.1 pmol of each TAG molecular species from chloroform extracts and is linear over a 1000-fold dynamic range. The sensitivity of individual TAG molecular species to ESI/MS/MS analyses correlated with the unsaturation index and inversely correlated with total aliphatic chain length of TAG. An algorithm was developed which identifies sensitivity factors, thereby allowing the rapid quantitation and molecular species fingerprinting of TAG molecular species directly from chloroform extracts of biological samples.
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Fatty acids detected through the TAG parent ion-neutral loss relationship represent esterified fatty acyl chains within TAG rather than free fatty acids.
“Thus, fatty acids detected through this parent ion -neutral loss relationship represent esteri fi ed fatty acyl chains within TAG rather than free fatty acids 78 .”
From APOE2 attenuates APOE4-associated amyloid pathology and related toxicity in a mouse model of Alzheimer’s disease (2026) · cites this paper as [78]
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Triglyceride accumulation protects against fatty acid-induced lipotoxicity
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- Authors
- Han, X., & Gross, R. W.
- Affiliations
- Washington University in St. Louis (US)
- Corresponding author
- Han, Xianlin
- Venue
- Analytical Biochemistry
- Publisher
- Elsevier BV
- Published
- 2001-08
- Volume
- 295
- Issue
- 1
- Pages
- 88-100
- ISSN
- 0003-2697 (ISSN-L)
- OpenAlex
- W1984044242
- Cited by
- 358
- References
- 1
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 7 · 2023: 10 · 2022: 12 · 2021: 13 · 2020: 17 · 2019: 10 · 2018: 14 · 2017: 13 · 2016: 19 · 2015: 19 · 2014: 22 · 2013: 26 · 2012: 32
- FWCI
- 4.36
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Analytical Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01aa011094-02)
- U.S. Public Health Service (JDFI 996003)
- National Institute on Alcohol Abuse and Alcoholism (R01 AA11094)
- National Heart, Lung, and Blood Institute (P01HL057278, R02 HL 41250)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
- Keywords
- triglycerides, choline glycerophospholipids, neutral loss scanning, fatty acid identification, sensitivity coefficients, unsaturation index, aliphatic chain length, lipidomics
- MeSH terms
- Animals, Lipids, Male, Myocardium, Triglycerides, Rats, Sprague-Dawley, Spectrometry, Mass, Electrospray Ionization, Rats
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