Laboratory of Toxicology, Department of Bioanalysis, Faculty of Pharmaceutical Sciences, Ghent University, Ottergemsesteenweg 460, 9000, Ghent, Belgium.
Department of Pediatric Intensive Care, Ghent University Hospital, De Pintelaan 185, 9000, Ghent, Belgium.
Anal Bioanal Chem. 2019 Jan;411(1):181-191. doi: 10.1007/s00216-018-1427-6. Epub 2018 Oct 23.
In the field of bioanalysis, dried matrix spot sampling is increasingly receiving interest, as this alternative sampling strategy offers many potential benefits over traditional sampling, including matrix volume-sparing properties. By using a microsampling strategy, e.g., volumetric absorptive microsampling (VAMS), the number of samples that can be collected from a patient can be increased, as a result of the limited sample volume that is required per sample. To date, no VAMS-based methods have been developed for the quantification of analytes in cerebrospinal fluid (CSF). The objective of this study was to develop and validate two LC-MS/MS methods for the quantification of paracetamol in dried blood and dried CSF, with both matrices sampled using VAMS. Both methods were fully validated based on internationally accepted guidelines. Paracetamol was chromatographically separated from its glucuronide and sulfate metabolites and no carry-over or unacceptable interferences were detected. The total precision (%RSD) was below 15% for all QC levels and accuracy (%bias) was below 7% (17% for the LLOQ of aqueous VAMS). The influence of the hematocrit on the recovery of blood VAMS samples appeared to be limited within the hematocrit range of 0.21 to 0.62. The blood VAMS samples were stable for 1 week if stored at 50 °C, and for at least 8 months when stored between - 80 °C and room temperature. The aqueous VAMS samples were stable for at least 9 months when stored between - 80 and 4 °C, and for 1 month when stored at room temperature. Application of the methods on external quality control material and analysis of patient samples demonstrated the validity and utility of the methods and provided a proof of concept for the analysis of CSF microsamples obtained via VAMS devices. Graphical abstract ᅟ.
在生物分析领域，干燥基质点采样越来越受到关注，因为这种替代采样策略相对于传统采样具有许多潜在的优势，包括节省基质体积的特性。通过使用微采样策略，例如体积吸收微采样（VAMS），可以增加从患者中采集的样本数量，因为每个样本所需的样本量有限。迄今为止，尚未开发基于 VAMS 的方法来定量分析脑脊液（CSF）中的分析物。本研究的目的是开发和验证两种 LC-MS/MS 方法，用于定量测定干燥血液和干燥 CSF 中的对乙酰氨基酚，这两种基质均采用 VAMS 采样。两种方法均根据国际公认的指南进行了全面验证。对乙酰氨基酚与葡萄糖醛酸和硫酸盐代谢物在色谱上分离，未检测到交叉污染或不可接受的干扰。所有 QC 水平的总精密度（%RSD）均低于 15%，准确度（%偏倚）均低于 7%（水性 VAMS 的LLOQ 为 17%）。在 0.21 至 0.62 的血细胞比容范围内，血细胞比容对血液 VAMS 样本回收率的影响似乎有限。如果在 50°C 下储存，血液 VAMS 样本可稳定 1 周，如果在-80°C 至室温之间储存，则可稳定至少 8 个月。如果在-80 和 4°C 之间储存，水性 VAMS 样本可稳定至少 9 个月，如果在室温下储存，则可稳定 1 个月。方法在外部质量控制材料上的应用和对患者样本的分析证明了方法的有效性和实用性，并为通过 VAMS 装置获得的 CSF 微样本分析提供了概念验证。
