Dried Blood Spots as a Sample Collection Technique for the Determination of Pharmacokinetics in Clinical Studies: Considerations for the Validation of a Quantitative Bioanalytical Method
Dried Blood Spots as a Sample Collection Technique for the Determination of Pharmacokinetics in Clinical Studies: Considerations for the Validation of a Quantitative Bioanalytical Method
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DOI：
10.1021/ac8022839
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发表时间：
2009-02-15
影响因子：
7.4
通讯作者：
Barfield, Matt
中科院分区：
文献类型：
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作者：
Spooner, Neil;Lad, Rakesh;Barfield, Matt
在临床研究中，利用纸上的干血点（DBS）而不是传统的血浆样本，开发了一种新的方法来定量测定循环药物浓度。使用对乙酰氨基酚作为工具化合物（范围为25至5000 ng/mL人血），验证了需要小血容量（15 μ L）的定量生物分析hplc -MS/MS分析。该方法采用简单的溶剂萃取DBS样品，然后采用反相高效液相色谱分离，结合选择性反应监测质谱检测。除了进行常规实验以建立符合国际公认标准（精密度、准确度、线性度、灵敏度、选择性）的分析方法的有效性外，还进行了一些实验来具体证明使用这种新型样品格式生成的定量数据的质量，即分析物和代谢物在全人血液和DBS样品中的稳定性；斑点血的量、用来斑点血的装置或斑点血的温度的影响。经验证的DBS方法已成功应用于临床研究（单次口服500mg或1g对乙酰氨基酚）。
A novel approach has been developed for the quantitative determination of circulating drug concentrations in clinical studies using dried blood spots (DBS) on paper, rather than conventional plasma samples. A quantitative bioanalytical RPLC-MS/MS assay requiring small blood volumes (15 mu L) has been validated using acetaminophen as a tool compound (range 25 to 5000 ng/mL human blood). The assay employed simple solvent extraction of a punch taken from the DBS sample, followed by reversed phase HPLC separation, combined with selected reaction monitoring mass spectrometric detection. In addition to performing routine experiments to establish the validity of the assay to internationally accepted criteria (precision, accuracy, linearity, sensitivity, selectivity), a number of experiments were performed to specifically demonstrate the quality of the quantitative data generated using this novel sample format, namely, stability of the analyte and metabolites in whole human blood and in DBS samples; effect of the volume of blood spotted, the device used to spot the blood, or the temperature of blood spotted. The validated DBS approach was successfully applied to a clinical study (single oral dose of 500 mg or 1 g acetaminophen).