Abstract：
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 (r2) 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.
摘要：
干血斑（DBS）彻底改变了使用LC-MS的治疗药物监测，用于精确定量心血管药物（CD），能够以最小的侵入性适应患者特异性药代动力学的个性化治疗。本研究旨在实现DBS中8种CD的同时定量。克服物理化学挑战。采用两步蛋白沉淀法进行简单、精确的样品制备。使用LC-MS/MS在ESI正离子模式下分析药物，显示高灵敏度和线性，相关系数（r2）超过0.999，在反相色谱上通过梯度洗脱含有0.1％甲酸2mM甲酸铵的DW-乙腈进行分离后。验证结果表明选择性好,没有观察到的基质效应和结转。大多数药物的日内和日间准确度和精确度在6%以内，除了低治疗水平的地高辛和地羊角苷，差异在20％以内。稳定性试验证实了合适的DBS处理和储存条件,表明药物在室温下至少30天的稳定性。对整个斑点的分析在所有目标药物中都证明了显着的准确性和可靠性。3毫米内径圆盘的分析，进出星展银行，推测含有3微升血液,对大多数药物显示出可接受的准确性，地高辛和地兰苷等极性较低的药物显示出较低的准确性，表明由于药物分布不均匀，需要进一步校正。因此,开发的LC-MS/MS方法可以在单个DBS分析中定量多个CD，同时暗示了基于准确性分析的潜力。
