Institute of Analytical Chemistry of the Czech Academy of Sciences, Veveří 97, CZ-60200 Brno, Czech Republic.
Faculty of Science, Department of Chemistry, Masaryk University, Kamenice 5, CZ-62500 Brno, Czech Republic.
Anal Chem. 2023 Aug 8;95(31):11823-11830. doi: 10.1021/acs.analchem.3c02474. Epub 2023 Jul 28.
A novel concept for highly versatile automated analyses of dried blood spot (DBS) samples by commercial capillary electrophoresis (CE) is presented. Two interchangeable CE cartridges with different fused-silica capillaries were used for the DBS elutions and the DBS eluate analyses, respectively. The application of one CE cartridge with a wide-bore capillary reduced DBS processing times to a minimum (1-2 min per sample) while fitting the other CE cartridge with a narrow-bore capillary served for highly efficient CE analyses. A comprehensive investigation of major variables affecting liquid handling in CE (capillary length, internal diameter, and temperature) was carried out with the aim of optimizing both procedures and enabling their maximum flexibility. The application of two CE cartridges also enabled the employment of CE detectors with different instrumental set-ups and/or principles as was demonstrated by the optical detection of nonsteroidal anti-inflammatory drugs (NSAIDs) and the conductivity detection of amino acids (AAs). The presented methods were optimized for the automated CE analyses of 36 DBS samples formed by a volumetric collection of 5 μL of capillary blood onto Whatman 903 discs and processed by direct in-vial elution using the CE instrument. The precision of liquid transfers for the automated DBS elutions was better than 0.9% and the precision of CE analyses did not exceed 5.1 and 12.3% for the determination of NSAIDs and AAs, respectively. Both methods were linear ( ≥ 0.996) over the therapeutic (NSAIDs) and the endogenous (AAs) concentration ranges, had limits of quantification below the typical analyte concentrations in human blood, and achieved sample throughputs of more than 6 DBSs per hour.
提出了一种新颖的概念,用于通过商业毛细管电泳(CE)对干血斑(DBS)样品进行高度通用的自动化分析。使用两个可互换的 CE 卡盒,分别用于 DBS 洗脱和 DBS 洗脱液分析,其中一个 CE 卡盒带有宽口径毛细管,将 DBS 处理时间缩短至最短(每个样品 1-2 分钟),而另一个 CE 卡盒带有细口径毛细管,可进行高效 CE 分析。对影响 CE 中液体处理的主要变量(毛细管长度、内径和温度)进行了全面研究,目的是优化这两个过程并使其具有最大的灵活性。使用两个 CE 卡盒还可以使用具有不同仪器设置和/或原理的 CE 检测器,如非甾体抗炎药(NSAIDs)的光学检测和氨基酸(AA)的电导率检测。所提出的方法针对通过将 5μL 毛细管血体积收集到 Whatman 903 盘上形成的 36 个 DBS 样品的自动化 CE 分析进行了优化,并通过 CE 仪器直接在管内洗脱进行处理。自动 DBS 洗脱的液体转移精度优于 0.9%,CE 分析的精度对于 NSAIDs 和 AAs 的测定分别不超过 5.1%和 12.3%。两种方法均具有线性(≥0.996),治疗(NSAIDs)和内源性(AA)浓度范围,定量下限低于人血中典型分析物浓度,并且每小时可处理超过 6 个 DBS 样品。