Electrochemically-Controlled Rapid Chromatographic Separation of Nuisance Compounds from Natural Product Extracts

摘要 天然产物仍然是药物发现化合物的丰富来源。然而，一个重大挑战是， 针对分子靶标的高通量生物测定筛选是分离生物活性化合物混合物 不含干扰性有害化合物。植物中普遍存在的有害化合物 提取物是单宁-多酚类植物代谢产物。固相萃取（SPE）工艺目前正获得 公认为一种快速分离植物粗提取物的方法， 化合物以及在将分析物引入更昂贵的气体或 液相色谱仪我们的客户发现访谈发现， 天然产物提取过程中去除有害化合物的现有反相材料 包括不适于单次使用、pH（在碱性pH下降解）和温度（在T > 60°C）的限制，以及在去除有害化合物如单宁方面的次优性能。这些 技术问题影响（增加）了生产该产品的运营成本，并降低了利润率。 SBIR第二阶段提案的总体目标是完成一个创新的固相萃取装置， 包含导电的全碳球形微粒（直径40 µM），使用 多壁碳纳米管（MWCNT）作为起始材料。这种材料在外加电压下， 有利于单宁的选择性分离和检测。在第二阶段，将实现两个目标。目标1. 证明全碳微珠合成和固相萃取基质是可扩展的。目的 2.在按比例放大的微珠和底物批次中建立一致的性能，并证明 (a)改进的单宁保留，（B）通过电喷雾质谱法快速检测捕获的单宁， 以及（c）重新使用所述筒。 预期成果包括：（1）最后确定能够大规模实施的设置和条件 全碳固相萃取系统的制造。2)关键合成工艺的确定 影响单宁提取性能再现性的参数（CPP），必须进行监测， 进行控制，以确保该过程产生所需的质量，而没有批次变异性。3)最后确定 科学合理且适当的检测方法，最终可用于批放行确认/验证 试验. II期活动的科学成果将促进该产品的商业推出 以及使其能够应用于去除其它有害化合物。在完成开发后， NanoPak-C将吸引寻求下一代性能能力的客户， 在天然产物提取过程中去除有害化合物的现有技术的能力， 从天然产物的纯化提取物中提前分离活性候选药物。

Abstract Natural products remain a rich source of compounds for drug discovery. However, a significant challenge for high throughput bioassay screening against molecular targets is the isolation of bioactive compound mixture free from interfering nuisance compounds. The well-established ubiquitous nuisance compounds in plant extracts are Tannins - polyphenolic plant metabolites. The solid-phase extraction (SPE) process is now gaining recognition as a method for rapid fractionation of crude plant extracts and employed to remove nuisance compounds as well as purify and concentrate analytes before introducing them into more expensive gas- or liquid- chromatography instrumentation. Our customer discovery interviews found that the challenges with current reverse phase materials in the removal of nuisance compounds during natural product extraction include unsuitable for more than single-use, pH (degrades at basic pH) and temperature (degrades at T > 60°C) restrictions, and suboptimal performance in the removal of nuisance compounds such as tannins. These technical issues affected (increased) the operational costs to manufacture the product and decreased margins. This SBIR Phase II proposal’s overall objective is to finalize an innovative solid phase extraction unit that incorporates electrically-conducting all-carbon spherical microparticles (40 µM diameter), synthesized using multi-walled carbon nanotubes (MWCNTs) as starting material. This material will, under an applied voltage, facilitate selective separation and detection of tannins. During Phase II, two aims will be pursued. Aim 1. Demonstrate that the all-carbon microbead synthesis and solid-phase extraction substrates are scalable. Aim 2. Establish consistent performance across the scaled-up microbead and substrate batches, and demonstrate (a) improved retention of tannins, (b) rapid detection of trapped tannins by electrospray mass spectrometry, and (c) reuse of the cartridge. Expected outcomes include 1) Finalization of the setup and conditions that will enable the large scale manufacturing of all-carbon solid-phase extraction system. 2) Identification of the critical synthesis process parameters (CPP) that affects tannin extraction performance reproducibility and has to be monitored and controlled to ensure the process produces the desired quality without batch variability. 3) Finalization of scientifically sound and appropriate test methods that can eventually be qualified/validated for batch release testing. The scientific outcomes from the Phase II activities will facilitate the commercial launch of this product as well as enable its application for the removal of other nuisance compounds. Upon complete development, NanoPak-C will attract customers seeking next-generation performance capabilities that fall outside the capabilities of current state-of-art for the removal of nuisance compounds during natural product extraction and advance separation of active drug candidates from the purified extracts of natural products.