1. A sensitive, expandable AQC-based LC-MS/MS method to measure amino metabolites and sphingolipids in cell and serum samples.
- Author
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Li X, Tian S, Riezman I, Qin Y, Riezman H, and Feng S
- Subjects
- Humans, Reproducibility of Results, Chromatography, High Pressure Liquid methods, Carbamates blood, Carbamates chemistry, Aminoquinolines chemistry, Aminoquinolines blood, Diabetes Mellitus, Type 2 blood, Diabetes Mellitus, Type 2 metabolism, Limit of Detection, Linear Models, Sensitivity and Specificity, Liquid Chromatography-Mass Spectrometry, Tandem Mass Spectrometry methods, Sphingolipids blood, Sphingolipids chemistry, Amino Acids blood, Amino Acids chemistry
- Abstract
Sphingolipids are a major lipid species found in all eukaryotes. Among structurally complex and diversified lipids, sphingoid bases have been heavily linked to various metabolic diseases. However, most current LC-MS-based methods lack the sensitivity to detect low-abundant sphingoid bases. The 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatization reagent, which efficiently forms covalent bonds with amino groups, has been widely used for amino acid detection. Nevertheless, the commonly used reverse-phase HPLC method for amino acid analysis is not suitable for amphipathic sphingolipids. To address this issue, we report a robust reverse-phase HPLC-MS/MS method capable of separating and detecting hydrophilic amino acids and sphingoid bases in a single run with high sensitivity. This method is also inclusive of other amino metabolites with an expandable target list. We tested this method under various conditions and samples, demonstrating its high reproducibility and sensitivity. Using this approach, we systematically analyzed human serum samples from healthy individuals, dyslipidemia, and type II diabetes mellitus (T2DM) patients, respectively. Two sphingolipids and five amino acids were identified with significant differences between the control and T2DM groups, highlighting the potential of this method in clinical studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Published
- 2024
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