Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China; Clinical Pharmacology Laboratory, Department of Pharmacy Practice and Science, University of Nebraska Medical Center, Omaha, NE, 68198, United States.
Clinical Pharmacology Laboratory, Department of Pharmacy Practice and Science, University of Nebraska Medical Center, Omaha, NE, 68198, United States.
J Pharm Biomed Anal. 2020 May 10;183:113147. doi: 10.1016/j.jpba.2020.113147. Epub 2020 Feb 4.
A rapid and sensitive liquid chromatographic-tandem mass spectrometric method was developed and validated to simultaneously quantitate vitamin K (PK) and vitamin K (MK-4) concentrations in human plasma to evaluate nutritional interventions. Charcoal-stripped human plasma was used for standard and calibration preparation with vitamin E-d6 as the internal standard. Patient plasma samples were extracted using n-hexane and analytes separated using an ACE -PFP C18 column (50 × 2.1 mm, 3 μ) equipped with standard C guard. The mobile phase consisted of 0.1 % formic acid in water and 0.1 % formic acid in methanol at a flow rate of 0.5 mL/min. Analyte quantification was achieved by using MS/MS with a positive atmospheric pressure chemical ionization in multiple reaction monitoring (MRM) mode. The lower limit of quantification was 0.01 ng/mL for both PK and MK-4. The assay was linear over the concentration range from 0.01-50 ng/mL for all analytes with a determination coefficient (r) of 0.998 or better. The intra-and inter-day accuracy and precision were within the acceptable limits per FDA guidance. The validated method was successfully applied to a clinical study for quantification of PK and MK-4 in human plasma to assess nutritional status and dietary interventions in patients with neurodegenerative diseases. Baseline plasma concentrations of PK and MK-4 ranged from 0.23 to 1.81 ng/mL and 0.33-0.57 ng/mL, respectively.
建立并验证了一种灵敏、快速的液相色谱-串联质谱法，用于同时定量人血浆中的维生素 K（PK）和维生素 K（MK-4）浓度，以评估营养干预措施。采用维生素 E-d6 作为内标，对经活性炭处理的人血浆进行标准和校准品制备。采用正己烷提取患者血浆样品，使用 ACE-PFP C18 柱（50×2.1mm，3μm）进行分离，标准 C 保护柱。流动相由 0.1%甲酸水溶液和 0.1%甲酸甲醇溶液组成，流速为 0.5mL/min。采用正大气压化学电离多反应监测（MRM）模式的 MS/MS 进行分析物定量。PK 和 MK-4 的定量下限均为 0.01ng/mL。所有分析物的浓度范围为 0.01-50ng/mL 时，该测定方法呈线性，相关系数（r）均大于 0.998。根据 FDA 指南，日内和日间精密度和准确度均在可接受范围内。该验证方法成功应用于一项临床研究，用于定量检测神经退行性疾病患者的人血浆中 PK 和 MK-4，以评估其营养状况和饮食干预措施。PK 和 MK-4 的基线血浆浓度分别为 0.23-1.81ng/mL 和 0.33-0.57ng/mL。
