Digital Microfluidic Sample Preparation of Blood for Mass Spectrometry

用于质谱分析的血液数字微流体样品制备

• 批准号: 8522972
• 负责人: Nick C Trotta
• 金额: $ 18.1万
• 依托单位: ADVANCED LIQUID LOGIC
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2014-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8522972
• 关键词: Adoption; Air; Amino Acids; Automation; Biochemical Genetics; Biological; Blood; Clinical; Clinical Research; Collaborations; Complex; Computer software; Corrosives; Development; Devices; Electronics; Goals; Human; Laboratories; Liquid substance; Logic; Maps; Mass Spectrum Analysis; Methods; Microfluidics; Neonatal Screening; Performance; Preparation; Price; Process; Protocols documentation; Reagent; Research; Research Personnel; Robotics; Sampling; Solvents; Spottings; System; Technology; Testing; Time; Universities; Work; acylcarnitine; base; cost; cost effective; design; digital; evaporation; flexibility; high throughput analysis; improved; innovation; instrument; interest; mass spectrometer; multiplex detection; novel; operation; programs; public health relevance; reconstitution; scale up; small molecule; succinylacetone

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.

描述：质谱仪是一种用于新生儿筛查和临床实验室的高通量生物分子分析的多路技术。在成熟的MS方法中，样品制备的复杂性被认为是主要的瓶颈。数字微流控是一个快速发展的领域，它利用基于微滴的液体处理。自.以来 高水平的自动化和更少的液体体积是该技术固有的，数字微流体是唯一适合高通量MS样品制备的产品。这项提议的目标是开发一种新的数字微流控平台，用于在引入质谱仪之前处理干血斑样本。样品制备所需的所有步骤都将在色谱柱上进行，包括分析物提取、添加适当的内标和样品衍生化。将首次开发新的盒上功能，如溶剂挥发和液-液提取。具体目标包括(I)实施提取和衍生化协议所需的新型电润湿能力，(Ii)使用现有的8个样品盒展示放大能力，并开发特定用途的数字微流控盒和仪器，以及(Iii)展示新生儿从干血斑中筛选氨基酸、酰肉碱和琥珀酸丙酮的样品制备。这项工作将与杜克大学的调查人员合作进行。拟议的平台将减少当前质谱学工作流程的复杂性、人为错误、成本和时间，从而提供更有效和更精确的生物分析测试。