建立并验证一种同时测定 25(OH)D3、epi-25(OH)D3、25(OH)D2、维生素 A、α-生育酚和 γ-生育酚的液相色谱串联质谱法。
Development and Validation of a Liquid Chromatography Mass Spectrometry Method for Simultaneous Measurement of 25(OH)D3, epi-25(OH)D3, 25(OH)D2, Vitamin A, α-Tocopherol, and γ-Tocopherol.
机构信息
Department of Pathology and Laboratory Medicine, Children's Hospital of Los Angeles, Los Angeles, CA, USA.
Department of Clinical Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
出版信息
Am J Clin Pathol. 2022 Jan 6;157(1):15-22. doi: 10.1093/ajcp/aqab088.
OBJECTIVES
Fat-soluble vitamins are measured to identify deficiencies that may lead to rickets, osteomalacia, night blindness, and reversible motor and sensory neuropathies. We present a rapid liquid chromatography-mass spectrometry (LC-MS/MS) method that simultaneously measures 25-hydroxyvitamin D3 (25[OH]D3), epi-25(OH)D3, 25(OH)D2, vitamin A, α-tocopherol, and γ-tocopherol.
METHODS
We mixed 100 µL serum with internal standard and extracted it by using supported liquid extraction plates. Reconstituted specimens were analyzed by LC-MS/MS with a 10-minute gradient.
RESULTS
The method was linear, covering physiological levels with r2 > 0.99, and the total precision was less than 15% at all quality control levels. The lower limit of the measuring intervals for 25(OH)D3, epi-25(OH)D3, 25(OH)D2, vitamin A, α-tocopherol, and γ-tocopherol were 4 ng/mL, 4 ng/mL, 4 ng/mL, 1 µg/dL, 0.2 µg/mL, and 0.2 µg/mL, respectively, with coefficient of variation of 20% or less. Recoveries were between 92% and 111% for National Institute of Standards and Technology reference materials and 81% and 122% for spike-recovery studies. Comparison studies for vitamin D total, vitamin A, and α-tocopherol demonstrated slopes between 1.04 and 1.11 and r2 between 0.94 and 0.96. Minimal matrix effect was observed for all analytes.
CONCLUSIONS
We developed and validated a rapid LC-MS/MS method for the simultaneous measurement of 25(OH)D3, epi-25(OH)D3, 25(OH)D2, vitamin A, α-tocopherol, and γ-tocopherol.
目的
检测脂溶性维生素，以发现可能导致佝偻病、骨软化症、夜盲症和可逆性运动及感觉神经病的维生素缺乏症。我们介绍一种快速液相色谱-质谱联用法（LC-MS/MS），该方法可同时测定 25-羟维生素 D3（25[OH]D3）、表-25（OH）D3、25（OH）D2、维生素 A、α-生育酚和 γ-生育酚。
方法
我们将 100 μL 血清与内标混合，并用固相萃取板进行提取。再用 10 分钟梯度洗脱，对经重溶的标本进行 LC-MS/MS 分析。
结果
该方法呈线性，r2＞0.99，涵盖了生理水平，所有质控水平的总精密度均＜15%。25（OH）D3、表-25（OH）D3、25（OH）D2、维生素 A、α-生育酚和 γ-生育酚的检测下限分别为 4ng/mL、4ng/mL、4ng/mL、1μg/dL、0.2μg/mL 和 0.2μg/mL，变异系数均＜20%。国家标准化与技术研究院参考物质的回收率在 92%～111%之间，加标回收率研究的回收率在 81%～122%之间。维生素 D 总量、维生素 A 和 α-生育酚的比较研究显示，斜率在 1.04～1.11 之间，r2 在 0.94～0.96 之间。所有分析物的基质效应均较小。
结论
我们建立并验证了一种快速 LC-MS/MS 方法，用于同时测定 25（OH）D3、表-25（OH）D3、25（OH）D2、维生素 A、α-生育酚和 γ-生育酚。
Zotero 插件