Automated technology for purification of active ingredients in natural products

描述（由申请人提供）：在涉及天然产物和其他复杂生物源样品的生物医学研究中，复杂天然混合物的分析是一项重大的科学挑战，特别是在需要分离次要成分时。逆流色谱（CCC）提供高负载能力，完整的样品回收，和高纯度的分数。它利用了一种温和的液体固定相，防止不稳定样品的降解，并且已经是天然产物研究中的一种有价值的工具。CCC分离消耗相对较少的溶剂，不需要使用昂贵的固体支撑，使其在环境和财政上都非常经济。耗材的节约使用使得CCC比其他竞争技术更加“绿色”。划分系数（k值）的测定是CCC提供的一项独特的分析能力。然而，用现代CCC仪器精确测量分析物分配系数要求样品负载远低于系统容量。在目前的技术条件下，当确定K值的条件优化时，CCC的独特实用优势基本上就丧失了。此外，目前成功使用CCC需要专门的培训和大量的操作时间。自动化，就像它对许多其他技术一样，可以有效地减轻学术负担，即吸收散布在数百篇期刊文章中数十年的关键信息，以及操作复杂的多组件系统的实际负担。液态CCC柱的动态特性所带来的挑战阻碍了自动化CCC的发展，直到SBC在SBIR项目的第一阶段取得了突破性的概念验证。该项目的长期目标，以及最终的市场产品，将是一个功能齐全的自动化CCC控制器，它将为最低限度的培训用户提供CCC的分离能力，并增强当前的能力，并为有经验的用户提供新的功能。自动化CCC将为美国国立卫生研究院（NIH）的目标做出重大贡献，使生物医学研究人员能够分析复杂的生物活性样品并减轻人类疾病。该项目的成功将通过将其应用于通常食用的植物产品中存在的生物活性天然产物的纯化和研究来证明，其中一种已在防治结核病方面显示出希望。

DESCRIPTION (provided by applicant): The analysis of complex natural mixtures presents a significant scientific challenge in biomedical studies involving natural products and other complex biogenic samples, particularly when the need for isolation of minor constituents arises. Countercurrent chromatography (CCC) offers high loading capacity, complete sample recovery, and high purity fractions. It makes use of a gentle liquid stationary phase that prevents degradation of labile samples, and is already a valuable tool in natural products research. CCC separations consume relatively little solvent and don't require the use of expensive solid supports, making them highly economical, both environmentally and fiscally. The frugal use of consumables makes CCC much more "green" than other competing technologies. Determination of partition coefficients (K-values) is a unique analytical capability offered by CCC. However, accurate measurement of analyte partition coefficients with modern CCC instruments requires that sample loading be constrained far below system capacity. With current technology, the unique practical advantages of CCC are essentially lost when the conditions are optimized to determine K- values. In addition, successful use of CCC currently requires specialized training and considerable operator time. Automation, as it has for many other technologies, could effectively mitigate both the academic burden of assimilating decades of key information spread over hundreds of journal articles, and the practical burden of operating a complex multi-component system. Challenges imposed by the dynamic nature of the liquid CCC column had prevented the development of automated CCC until the SBC achieved a breakthrough proof of concept during Phase I of this SBIR project. The long-term objective of this project, and the ultimately marketable product, will be a fully functional automated CCC controller that will provide the separation power of CCC to minimally trained users, as well as enhance current capabilities and provide new functionality to experienced users. Automated CCC will provide a significant contribution to the goals of the National Institutes of Health (NIH), empowering biomedical researchers in their quest to analyze complex bioactive samples and alleviate human disease. The success of this project will be demonstrated by its application to the purification and study of bioactive natural products present in commonly consumed botanical products, one of which has shown promise in the fight against tuberculosis. PUBLIC HEALTH RELEVANCE: The analysis of highly complex chemical mixtures presents a significant scientific challenge, particularly in biomedical studies involving natural products and other complex biogenic samples. Countercurrent chromatography (CCC) is a technique, originally developed at NIH, which is able to solve many challenges in this area. This SBIR research proposal provides the key piece of technology required to bring the immense potential of CCC's analytical capabilities to realization, empowering biomedical researchers with a much-needed new tool in the arsenal to combat disease through biochemical discovery.