Li Fumin, Zulkoski John, Fast Douglas, Michael Steve
Covance Laboratories, Inc., Bioanalytical Chemistry, 3301 Kinsman Boulevard, Madison, WI 53704, USA.
Bioanalysis. 2011 Oct;3(20):2321-33. doi: 10.4155/bio.11.219.
Dried blood spots (DBS) in their current format encounter challenges in bioanalysis using fixed areas, including but not limited to, waste of DBS samples (only a fraction is used for analysis), the need for sample punching leading to concerns of sample carryover, uncertainty for accurate recovery assessments and hematocrit (HCT) effects. Here we describe a novel concept, namely perforated dried blood spots (PDBS), for accurate microsampling that addresses previous challenges. PDBS discs were prepared from regular filter paper, with a diameter of 6.35 mm and a thickness of 0.83 mm. An accurate amount of blood sample (5-10 µl), was deposited, dried and stored on the PDBS discs. Upon sample analysis, PDBS samples are simply pushed by single-use pipette tips into 96-well plates. The proof-of-concept study was carried out on a PDBS LC-MS/MS assay development and validation under GLP criteria for the quantitation of lansoprazole in human whole-blood (K(3)EDTA). Particularly, the effect of HCT on the accuracy of quantitation was found to be related to recovery from PDBS samples. In all, PDBS was proved to be a viable alternative to conventional DBS, offering additional advantages of complete sample utilization, no requirement for punching, ease of recovery assessments, and elimination of sampling influence due to HCT levels.
当前形式的干血斑（DBS）在使用固定区域进行生物分析时面临挑战，包括但不限于DBS样本的浪费（仅一小部分用于分析）、需要进行样本打孔从而引发对样本残留的担忧、准确回收率评估的不确定性以及血细胞比容（HCT）的影响。在此，我们描述了一种新的概念，即穿孔干血斑（PDBS），用于准确的微量采样，解决了先前的挑战。PDBS圆盘由常规滤纸制备而成，直径为6.35毫米，厚度为0.83毫米。将准确量的血样（5 - 10微升）滴加、干燥并储存在PDBS圆盘上。在进行样本分析时，只需用一次性移液器吸头将PDBS样本推至96孔板中。在GLP标准下，针对人全血（K(3)EDTA）中兰索拉唑的定量，开展了一项关于PDBS液相色谱 - 串联质谱（LC - MS/MS）分析方法开发与验证的概念验证研究。特别地，发现HCT对定量准确性的影响与从PDBS样本中的回收率有关。总体而言，PDBS被证明是传统DBS的可行替代方法，具有样本完全利用、无需打孔、回收率评估简便以及消除HCT水平对采样影响等额外优势。
