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<p>Accurate quantification of bioactive compounds in plant-derived extracts is essential for their characterization and quality control. In this study, an LC–MS/MS method was developed and applied for the determination of coumarin in <i>Melilotus albus</i> extract. The method demonstrated excellent linearity over the concentration range of 15–1000 ng/mL (R² = 0.996), with high sensitivity and selectivity under multiple reaction monitoring (MRM) conditions. Coumarin was successfully identified and quantified in <i>M. albus</i> extract at a concentration of 2.14 ± 0.18&#xa0;mg/g extract, confirming its suitability as a marker compound for extract characterization. In addition, the biological relevance of the characterized extract was evaluated in an in vitro model of diabetes-associated muscle atrophy using differentiated C2C12 myotubes exposed to hyperglycemic (33 mM) and glucocorticoid stress (dexamethasone, 10 µM). Treatment with <i>M. albus</i> extract improved cell viability and preserved myotube morphology. Mechanistically, the extract suppressed the upregulation of Forkhead box O3a (FoxO3a) and muscle RING finger protein-1 (MuRF-1) while restoring myogenic regulatory factors, including MyoD and myogenin. Overall, this study demonstrates that the developed LC–MS/MS method provides a reliable and sensitive approach for the quantification of coumarin in <i>M. albus</i> extract and supports its application in the analytical characterization of plant-derived matrices.</p>

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LC–MS/MS-based quantification of coumarin in Melilotus albus extract and its protective effects against diabetes-associated myotube atrophy

  • 期刊论文

  • 发表日期：

    2026年6月18日

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  • Eun-Chae Cho,
  • Seongeon Lee,
  • Yean Jung Choi

摘要

Accurate quantification of bioactive compounds in plant-derived extracts is essential for their characterization and quality control. In this study, an LC–MS/MS method was developed and applied for the determination of coumarin in Melilotus albus extract. The method demonstrated excellent linearity over the concentration range of 15–1000 ng/mL (R² = 0.996), with high sensitivity and selectivity under multiple reaction monitoring (MRM) conditions. Coumarin was successfully identified and quantified in M. albus extract at a concentration of 2.14 ± 0.18 mg/g extract, confirming its suitability as a marker compound for extract characterization. In addition, the biological relevance of the characterized extract was evaluated in an in vitro model of diabetes-associated muscle atrophy using differentiated C2C12 myotubes exposed to hyperglycemic (33 mM) and glucocorticoid stress (dexamethasone, 10 µM). Treatment with M. albus extract improved cell viability and preserved myotube morphology. Mechanistically, the extract suppressed the upregulation of Forkhead box O3a (FoxO3a) and muscle RING finger protein-1 (MuRF-1) while restoring myogenic regulatory factors, including MyoD and myogenin. Overall, this study demonstrates that the developed LC–MS/MS method provides a reliable and sensitive approach for the quantification of coumarin in M. albus extract and supports its application in the analytical characterization of plant-derived matrices.