Scholarlink

<p>Fat-soluble vitamins (A, D, and E) are essential micronutrients whose absorption critically depends on bile acid–mediated solubilization. However, studies associating bile acids and these vitamins remain limited, partly due to methodological challenges. Therefore, we developed a novel and rapid LC-MS/MS method for the simultaneous quantification of these vitamins in serum, based on 2-nitrosopyridine (PyrNo) click derivatization. This strategy employs Diels-Alder cycloaddition for vitamin A/D and acid-catalyzed oxidative dehydrogenation for vitamin E, enabling one-step protein precipitation followed by a 15-min room-temperature reaction. The method significantly enhanced sensitivity (&gt;350-fold for vitamin D), allowing analysis within 6&#xa0;min, and demonstrated excellent performance: linearity (<i>R</i> &gt; 0.99), accuracy (96.1–110.2%), precision (RSD ≤ 4.9%), with lower limits of quantification of 0.1&#xa0;µmol/L for retinol, 2&#xa0;nmol/L for 25-hydroxyvitamin D₂ and D₃, and 2&#xa0;µmol/L for α-tocopherol. Application of this method to a clinical cohort of 277 patients, covering a broad spectrum of total bile acid (TBA) concentrations, revealed a previously unrecognized biphasic relationship between TBA and vitamin status. Within the normal TBA group (≤10&#xa0;µmol/L), vitamin A deficiency was associated with lower median TBA, and a TBA cutoff of 3.7&#xa0;µmol/L showed predictive ability for combined vitamin A/E deficiency (AUC = 0.6057). Importantly, when patients with elevated TBA (&gt;10&#xa0;µmol/L) were included, vitamin A/D levels peaked in the intermediate TBA range (3.7–10&#xa0;µmol/L) and were lower in both the low (&lt;3.7&#xa0;µmol/L) and high (&gt;10&#xa0;µmol/L) TBA groups. This work provides both a major analytical advancement and novel clinical insights into bile acid–mediated vitamin homeostasis.</p> Graphical abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

A rapid LC-MS/MS method with 2-nitrosopyridine derivatization to determine fat-soluble vitamins, revealing a biphasic bile acids association in clinical cohorts

  • 期刊论文

  • 发表日期：

    2026年5月29日

查看全文
  • Fengli Jiang,
  • Yuting Gu,
  • Fan Yu,
  • Yan Liang,
  • Shuangshuang Yang,
  • Meiwei Zhang,
  • Can Xiao,
  • Meixian Ou,
  • Yang Xu,
  • Gang Li,
  • Chen Yu,
  • Gangyi Liu,
  • Jingying Jia,
  • Youli Lu

摘要

Fat-soluble vitamins (A, D, and E) are essential micronutrients whose absorption critically depends on bile acid–mediated solubilization. However, studies associating bile acids and these vitamins remain limited, partly due to methodological challenges. Therefore, we developed a novel and rapid LC-MS/MS method for the simultaneous quantification of these vitamins in serum, based on 2-nitrosopyridine (PyrNo) click derivatization. This strategy employs Diels-Alder cycloaddition for vitamin A/D and acid-catalyzed oxidative dehydrogenation for vitamin E, enabling one-step protein precipitation followed by a 15-min room-temperature reaction. The method significantly enhanced sensitivity (>350-fold for vitamin D), allowing analysis within 6 min, and demonstrated excellent performance: linearity (R > 0.99), accuracy (96.1–110.2%), precision (RSD ≤ 4.9%), with lower limits of quantification of 0.1 µmol/L for retinol, 2 nmol/L for 25-hydroxyvitamin D₂ and D₃, and 2 µmol/L for α-tocopherol. Application of this method to a clinical cohort of 277 patients, covering a broad spectrum of total bile acid (TBA) concentrations, revealed a previously unrecognized biphasic relationship between TBA and vitamin status. Within the normal TBA group (≤10 µmol/L), vitamin A deficiency was associated with lower median TBA, and a TBA cutoff of 3.7 µmol/L showed predictive ability for combined vitamin A/E deficiency (AUC = 0.6057). Importantly, when patients with elevated TBA (>10 µmol/L) were included, vitamin A/D levels peaked in the intermediate TBA range (3.7–10 µmol/L) and were lower in both the low (<3.7 µmol/L) and high (>10 µmol/L) TBA groups. This work provides both a major analytical advancement and novel clinical insights into bile acid–mediated vitamin homeostasis.

Graphical abstract