Development of targeted lipidomics analysis with accurate determination method and its clinical application

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K06507
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
• Research Institution: Fukuoka University
• Principal Investigator: Hayama Tadashi 福岡大学, 薬学部, 教授 (90549693)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ジアシルグリセロール / 液体クロマトグラフィー／質量分析計 / マトリックス効果 / リン脂質

Outline of Final Research Achievements

In this study, we developed an accurate method for determining diacylglycerols (DAGs) in human plasma using a fluorous solvent extraction method followed by LC-MS analysis. The lipid mixture in the plasma was extracted using the Bligh-Dyer method, and then, the resulting solution was subjected to fluorous solvent extraction to remove phospholipids via the specificity of fluorous affinity by forming a complex with a perfluoropolyethercarboxylic acid-lanthanum (Ⅲ) salt. The remaining DAGs in the non-fluorous solvent could be directly injected into the LC-MS/MS system. The removal rate of the phospholipids through the fluorous extraction was more than 99.9%; thus, the matrix effect-eliminating analysis of DAGs in human plasma with LC-MS was enabled. Furthermore, the applicability of this method and the possibility of using DAGs as biomarkers were demonstrated by applying this method to human plasma obtained from the major depressive disorder as a related disease.

Free Research Field

Analytical Chemistry

Academic Significance and Societal Importance of the Research Achievements

ジアシルグリセロール類は、種々の疾患との関連が報告されている。しかしながら、生体内には多種多様の脂質類が存在しており（とくにリン脂質の妨害を受けやすいこと）、必ずしも正確な測定を行うことができるとは限らない。そこで本研究では妨害となるリン脂質を選択的に除去することで、ジアシルグリセロール類のLC-MS定量における正確性を担保することとした。実際に、本法をうつ病患者から得られた臨床試料（血液）に適用したところ、その正確な定量値から健常人の有意差を見出すことが可能であった。今後、このような疾患のバイオマーカー測定法として早期診断や治療効果判定などに大きく貢献できることを期待している。