Opti-nQL: An Optimized, Versatile and Sensitive Nano-LC Method for MS-Based Lipidomics Analysis
Abstract
:1. Introduction
2. Results and Discussion
2.1. Method Development
Lipid Extraction and LC Optimization
2.2. Lipidomics Analysis of Mammalian Cells
2.2.1. Semi-Targeted Lipidomics Analysis
2.2.2. Untargeted Lipidomics Analysis
2.3. Opti-nQL Is Compatible with Alternative Lipid Extraction Methods
3. Materials and Methods
3.1. Materials
3.2. Biological Sample Collection and Cell Lysis
3.3. Proteins Extraction and Quantification
3.4. Lipids Extraction
3.5. Liquid Chromatography
3.6. Mass Spectrometry Analysis
3.7. Data Processing and Analysis
3.8. Statistics
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Class | Subclass | Species | Concentration |
---|---|---|---|
Glycero-Phospholipids | PG | PG 12:0/13:0 | 7.5 pmol |
PI | PI 12:0/13:0 | 54 pmol | |
PE | PE 12:0/13:0 | 52 pmol | |
PS | PS 12:0/13:0 | 43 pmol | |
PC | PC 12:0/13:0, | 40 pmol | |
Sphingolipids | CER | Ceramide d18:1/25:0 | 100 pmol |
GlcCer | GalCer d18:1/12:0 | 50 pmol | |
LacCer | LacCer d18:1/12:0 | 50 pmol | |
Sa | Sphinganine (d17:0) | 50 pmol | |
S | Sphingosine (d17:1) | 50 pmol | |
S1P | Sphingosine-1-P (d17:1) | 100 pmol | |
GalSph | Galactosyl(s) Sphingosine-d5 | 20 pmol | |
Glycerolipids | DAG | D5-DAG ISTD Mix I | 20 pmol |
TAG | D5-TAG ISTD Mix I | 20 pmol | |
Sterol Lipids | Chol | Chol-d7 | 800 pmol |
CE | CE (19:0) | 100 pmol |
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Cattaneo, A.; Martano, G.; Restuccia, U.; Tronci, L.; Bianchi, M.; Bachi, A.; Matafora, V. Opti-nQL: An Optimized, Versatile and Sensitive Nano-LC Method for MS-Based Lipidomics Analysis. Metabolites 2021, 11, 720. https://0-doi-org.brum.beds.ac.uk/10.3390/metabo11110720
Cattaneo A, Martano G, Restuccia U, Tronci L, Bianchi M, Bachi A, Matafora V. Opti-nQL: An Optimized, Versatile and Sensitive Nano-LC Method for MS-Based Lipidomics Analysis. Metabolites. 2021; 11(11):720. https://0-doi-org.brum.beds.ac.uk/10.3390/metabo11110720
Chicago/Turabian StyleCattaneo, Angela, Giuseppe Martano, Umberto Restuccia, Laura Tronci, Michele Bianchi, Angela Bachi, and Vittoria Matafora. 2021. "Opti-nQL: An Optimized, Versatile and Sensitive Nano-LC Method for MS-Based Lipidomics Analysis" Metabolites 11, no. 11: 720. https://0-doi-org.brum.beds.ac.uk/10.3390/metabo11110720