Department of Pharmacology & Therapeutics, Institute of Integrative, Systems & Molecular Biology, University of Liverpool, L7 8TX, UK.
Bioanalytical Facility, University of Liverpool, William Henry Duncan Building, Liverpool, L7 8TX, UK.
Bioanalysis. 2023 Jul;15(13):739-756. doi: 10.4155/bio-2023-0057. Epub 2023 Jun 9.
A rapid and sensitive LC-MS method has been developed and validated for the quantification of nucleoside di/triphosphates using a novel plasma separation card (HemaSep). Cards were spotted with whole blood and stored at -80°C. Metabolites were extracted using 70:30 MeOH:20% formic acid, followed by weak anion exchange SPE and eluted using a Biobasic-AX column. Quantification was performed using a triple quadrupole mass spectrometer with a calibration range of 1.25-250 pmol/sample. The recovery of metabolites was high (>93%). Precision and accuracy were acceptable and metabolites remained stable on the card after 29 days (stored at ambient temperature). HemaSep dried blood spots are a useful microsampling tool and offer an alternative to liquid plasma as they maintain stability over time.
已开发并验证了一种使用新型血浆分离卡（HemaSep）定量核苷二/三磷酸的快速灵敏 LC-MS 方法。将卡片点样全血并储存在-80°C。使用 70:30 MeOH:20%甲酸提取代谢物，然后进行弱阴离子交换 SPE 并使用 Biobasic-AX 柱洗脱。使用三重四极杆质谱仪进行定量，校准范围为 1.25-250 pmol/样品。代谢物的回收率高（>93%）。精密度和准确度可接受，并且在 29 天后（储存在环境温度下）卡上的代谢物仍保持稳定。HemaSep 干血斑是一种有用的微量采样工具，它们作为液体血浆的替代品提供了一种选择，因为它们随时间保持稳定。
