A sensitive, expandable AQC-based LC-MS/MS method to measure amino metabolites and sphingolipids in cell and serum samples

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1245; p. 124256
• Main Authors: Li, Xiaotian, Tian, Shuwei, Riezman, Isabelle, Qin, Yujiao, Riezman, Howard, Feng, Suihan
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.09.2024
• Subjects: amino acids; Amino metabolites; AQC; blood serum; derivatization; diabetes mellitus; Dyslipidemia; eukaryotic cells; humans; hydrophilicity; hyperlipidemia; LC-MS/MS; metabolites; reversed-phase high performance liquid chromatography; species; sphingolipids; surfactants; T2DM; LC-MS/MS; T2DM; AQC; Amino metabolites; sphingolipids; Dyslipidemia
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2024.124256

[Display omitted] •A robust reverse-phase HPLC-MS/MS method was proposed based on AQC derivatization.•This method is capable of separating and detecting hydrophilic amino acids and sphingoid bases in a single run with high sensitivity.•The method was applied to human serum samples for the screening of biomarkers.•Two sphingolipids and five amino acids were identified with significant differences between the control and T2DM groups. 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.