Organic Metrology Group, Division of Chemical and Material Metrology, Korea Research Institute of Standards and Science, Yuseong-gu, Daejeon 34113, Republic of Korea; Department of Bio-Analytical Science, University of Science and Technology, 217 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea; Department of Nutrition and Food Science, National Research Centre, Dokki, Cairo 12622, Egypt.
Organic Metrology Group, Division of Chemical and Material Metrology, Korea Research Institute of Standards and Science, Yuseong-gu, Daejeon 34113, Republic of Korea; Department of Bio-Analytical Science, University of Science and Technology, 217 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Jul 15;1242:124215. doi: 10.1016/j.jchromb.2024.124215. Epub 2024 Jun 22.
Dried Blood Spots (DBS) revolutionize therapeutic drug monitoring using LC-MS for the precise quantification of cardiovascular drugs (CDs), enabling personalized treatment adapted to patient-specific pharmacokinetics with minimal invasiveness. This study aims to achieve simultaneous quantification of eight CDs in DBS, overcoming physicochemical challenges. A two-step protein precipitation method was used for simple and precise sample preparation. The drugs were analyzed using LC-MS/MS in ESI positive-ion mode, showing high sensitivity and linearity, with a correlation coefficient (r) exceeding 0.999, after being separated on a reversed-phase chromatography by gradient elution of DW-acetonitrile containing 0.1 % formic acid + 2 mM ammonium formate. The validation results indicate good selectivity, with no observed matrix effect and carry-over. The intra- and inter-day accuracy and precision were within 6 % for most drugs, except for digoxin and deslanoside at low therapeutic levels where the variation was within 20 %. Stability tests confirmed suitable DBS handling and storage conditions, indicating drug stability for at least 30 days at room temperature. The analysis of whole spot has demonstrated remarkable precision and reliability in all target drugs. The analysis of 3 mm internal diameter discs, punched in and out of DBS, presumed to contain 3 µL of blood, showed acceptable accuracy for most drugs, with less polar drugs like digoxin and deslanoside showing lower accuracy, indicating a need for further correction due to non-uniform drug distribution. Consequently, the developed LC-MS/MS method enables the quantification of multiple CDs in a single DBS analysis, while suggesting the potential for accuracy-based analysis.
干血斑 (DBS) 通过 LC-MS 对心血管药物 (CDs) 进行精确定量，实现了治疗药物监测的革命，使个性化治疗适应于患者特定的药代动力学，具有最小的侵入性。本研究旨在实现 DBS 中八种 CDs 的同时定量，克服理化挑战。两步蛋白沉淀法用于简单而精确的样品制备。药物在 ESI 正离子模式下用 LC-MS/MS 分析，在反相色谱梯度洗脱 DW-乙腈中含有 0.1%甲酸+2mM 甲酸铵后，显示出高灵敏度和线性，相关系数 (r) 超过 0.999。验证结果表明具有良好的选择性，无观察到基质效应和交叉污染。大多数药物的日内和日间准确度和精密度在 6%以内，除了地高辛和去乙酰毛花苷在低治疗水平时变化在 20%以内。稳定性测试证实了 DBS 处理和储存条件的适宜性，表明药物在室温下至少稳定 30 天。全斑分析在所有目标药物中均表现出出色的精密度和可靠性。分析 3mm 内径的圆盘，从 DBS 中打孔和打孔外，假设包含 3µL 的血液，表明大多数药物的准确度可接受，而地高辛和去乙酰毛花苷等极性较小的药物准确度较低，表明由于药物分布不均匀，需要进一步校正。因此，所开发的 LC-MS/MS 方法能够在单个 DBS 分析中定量多种 CDs，同时表明了基于准确度的分析的潜力。
