Development of an advanced liquid chromatography–tandem mass spectrometry measurement system for simultaneous sphingolipid analysis

• Published in: Scientific Reports Vol. 14; no. 1; pp. 5699 - 13
• Main Authors: Uranbileg, Baasanjav, Sakai, Eri, Kubota, Masayuki, Isago, Hideaki, Sumitani, Masahiko, Yatomi, Yutaka, Kurano, Makoto
• Format: Journal Article
• Language: English
• Published: London Springer Science and Business Media LLC 08.03.2024; Nature Publishing Group UK; Nature Publishing Group; Nature Portfolio
• Subjects: 631/45; 631/61/320; Biological samples; Ceramide; Ceramide1-phosphate (Cer1P); Ceramides; Chromatography; Chromatography, Liquid; Deoxy-ceramide; dh-ceramide; Hexosylceramide (HexCer); Humanities and Social Sciences; Ionization; Laboratory tests; Lactosylceramide (LacCer); Liquid chromatography; Lysates; Mass spectrometry; Mass spectroscopy; Medicine; multidisciplinary; Q; R; Science; Science (multidisciplinary); Scientific imaging; Sphingolipid; Sphingolipids; Sphingolipids - metabolism; Sphingomyelin; Sphingomyelins; Sphingosine; Sphingosine 1-phosphate; Tandem Mass Spectrometry - methods; Hexosylceramide (HexCer); Deoxy-ceramide; Deoxy-dh-ceramide; Sphingolipid; dh-ceramide; Ceramide1-phosphate (Cer1P); Lactosylceramide (LacCer)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-56321-w

Mass spectrometry-based lipidomics approaches offer valuable tools for the detection and quantification of various lipid species, including sphingolipids. The present study aimed to develop a new method to simultaneously detect various sphingolipid species that applies to diverse biological samples. We developed and validated a measurement system by employing a single-column liquid chromatography-mass spectrometry system utilizing a normal-phase separation mode with positive ionization. The measurement system provided precision with a coefficient of variant below 20% for sphingolipids in all types of samples, and we observed good linearity in diluted serum samples. This system can measure the following sphingolipids: sphingosine 1-phosphate (S1P), sphingosine (Sph), dihydroS1P (dhS1P), dihydroSph (dhSph), ceramide 1-phosphate (Cer1P), hexosylceramide (HexCer), lactosylceramide (LacCer), dh-ceramide, deoxy-ceramide, deoxy-dh-ceramide, and sphingomyelin (SM). By measuring these sphingolipids in cell lysates where S1P lyase expression level was modulated, we could observe significant and dynamic modulations of sphingolipids in a comprehensive manner. Our newly established and validated measurement system can simultaneously measure many kinds of sphingolipids in biological samples. It holds great promise as a valuable tool for laboratory testing applications to detect overall modulations of sphingolipids, which have been proposed to be involved in pathogenesis processes in a series of elegant basic research studies.