Department of Drug Metabolism and Pharmacokinetics, Merck Research Laboratories, Summit, NJ 07901, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Nov 15;879(30):3626-38. doi: 10.1016/j.jchromb.2011.10.008. Epub 2011 Oct 11.
A rugged and robust liquid chromatographic tandem mass spectrometric (LC-MS/MS) method utilizing dried blood spots (DBS) was developed and validated for the analysis of posaconazole in human whole blood. Posaconazole fortified blood samples were spotted (15 μL) onto Ahlstrom Alh-226 DBS cards and dried for at least 2h. Punched spots were then extracted by using a mixture of acetonitrile and water containing stable labeled internal standard (IS). Posaconazole and its IS were separated from endogenous matrix components on a Kinetex™ C18 column under gradient conditions with a mobile phase A consisting of 0.1% formic acid and a mobile phase B consisting of 0.1% formic acid in acetonitrile/methanol (70/30, v/v). The analyte and IS were detected using a Sciex API 4000 triple quadrupole LC-MS/MS system equipped with a TurboIonSpray™ source operated in the positive ion mode. The assay was linear over the concentration range of 5-5000 ng/mL. The inter-run accuracy and precision of the assay were -1.8% to 0.8% and 4.0% to 10.4%, respectively. Additional assessments unique to DBS were investigated including sample spot homogeneity, spot volume, and hematocrit. Blood spot homogeneity was maintained and accurate and precise quantitation results were obtained when using a blood spot volume of between 15 and 35 μL. Human blood samples with hematocrit values ranging between 25% and 41% gave acceptable quantitation results. The validation results indicate that the method is accurate, precise, sensitive, selective and reproducible.
建立并验证了一种利用干血斑（DBS）的耐用、稳健的液相色谱-串联质谱（LC-MS/MS）方法，用于人全血中泊沙康唑的分析。将含有泊沙康唑的血样（15 μL）点样至 Ahlstrom Alh-226 DBS 卡上，干燥至少 2h。然后使用含稳定标记内标（IS）的乙腈和水的混合物提取点样。泊沙康唑及其 IS 在梯度条件下，于 Kinetex™ C18 柱上分离，流动相 A 为 0.1%甲酸，流动相 B 为乙腈/甲醇（70/30，v/v）中的 0.1%甲酸。采用 Sciex API 4000 三重四极杆 LC-MS/MS 系统，配有 TurboIonSpray™源，在正离子模式下进行检测。该分析物在 5-5000ng/mL 浓度范围内呈线性。分析的批内准确度和精密度分别为-1.8%至 0.8%和 4.0%至 10.4%。还对 DBS 特有的一些评估进行了研究，包括样本斑点均匀性、斑点体积和红细胞压积。当使用 15-35 μL 的血斑体积时，斑点均匀性得以维持，且可获得准确和精密度良好的定量结果。红细胞压积在 25%-41%之间的人血样本可获得可接受的定量结果。验证结果表明，该方法准确、精密、灵敏、选择性好且重现性佳。
