Digital Microfluidic Sample Preparation of Blood for Mass Spectrometry

描述：质谱是一种用于新生儿筛查和临床实验室中生物分子高通量分析的多重技术。样品制备的复杂性已被公认为成熟的MS方法的主要瓶颈。数字微流体是利用基于微滴的液体处理的快速发展的领域。以来 高水平的自动化和减少的液体体积是该技术的固有特点，数字微流体技术特别适合于高通量MS样品制备。该提案的目的是开发一种新的数字微流体平台，用于在引入质谱仪之前操纵干血斑样品。样品制备所需的所有步骤将在检测盒上进行，包括分析物提取、加入适当的内标物和样品衍生化。将首次开发新的柱上能力，如溶剂蒸发和液-液萃取。具体目标包括：（i）实现提取和衍生化方案所需的新型电润湿能力，（ii）使用现有的8样本盒和开发特定应用的数字微流体盒和仪器来演示放大能力，以及（iii）演示用于从干血点中新生儿筛选氨基酸，酰基肉毒碱和琥珀酰丙酮的样品制备。这项工作将与杜克大学的研究人员合作进行。拟议的平台将减少当前质谱工作流程的复杂性、人为错误、成本和时间，从而提供更有效和精确的生物分析测试。

DESCRIPTION: Mass spectrometry is a multiplex technology for high-throughput analysis of biomolecules in newborn screening and clinical laboratories. Complexity in sample preparation has been recognized as the main bottleneck in well-established MS methods. Digital microfluidics is a rapidly advancing field which utilizes microdroplet-based liquid handling. Since high levels of automation and reduced volumes of liquid are intrinsic to the technology, digital microfluidics is uniquely suitable for high-throughput MS sample preparation. The objective of this proposal is the development of a new digital microfluidic platform for manipulating dried blood spot samples prior to introduction into the mass spectrometer. All steps required for sample preparation will be implemented on cartridge, including analyte extraction, addition of appropriate internal standards, and sample derivatization. Novel on-cartridge capabilities such as solvent evaporation and liquid-liquid extraction will be developed for the first time. The specific aims include (i) implementation of novel electrowetting capabilities necessary for extraction and derivatization protocols, (ii) demonstration of scale up capabilities using an existing 8-sample cartridge and development of an application-specific digital microfluidic cartridge and instrument, and (iii) demonstration of sample preparation for newborn screening of amino acids, acylcarnitines and succinylacetone from dried blood spots. This work will be performed in collaboration with Duke University investigators. The proposed platform will reduce complexity, human error, costs and time of current mass spectrometry workflows, thereby providing more effective and precise bioanalysis tests.