Life sciences research and other critical bioanalytical applications would strongly benefit from faster and higher resolution liquid chromatographic (HPLC) separations for both small molecules and larger molecules such as proteins, peptides, glycopeptides, and glycans. Many HPLC separations of biomedical interest, such as proteomic or glycomic profiling, require hours or days for resolution of a single sample. Due to the complexity of biological samples, there are many examples in the current research literature that combine multiple dimensions of HPLC separations, often with online mass spectroscopy (MS). Although very time consuming, this approach has yielded useful information on protein expression and post-translational modifications, the biological processes underlying protein expression and modifications, and the effects thereon of development, disease state, and various environmental and genetic factors. In various complex bioanalytical workflows, high efficiency LC separations are a fundamental part of the analytical systems, and the time to achieve high resolution separations of complex biological samples is a great bottleneck. This proposal describes an approach to improve the separation efficiency of HPLC by extending our proprietary Fused-Core silica platform technology to smaller particles, providing significantly faster and higher resolution separations. Our objectives are to create 2.3 and 1.8 micron diameter superficially porous Fused-Core silica particles, and to load these materials efficiently into HPLC column formats. The goals of the proposed work would yield very high performance chromatographic products, greater than those currently available, to be applied broadly in analysis of complex biological samples, pharmaceutical and biopharmaceutical applications, in fact in any current application that uses HPLC methods. The separations technology described will directly lead to useful products for which there is a significant technical and market demand.

Public Health Relevance

High pressure liquid chromatography is the most widely used analytical method to separate mixtures of molecules, allowing measurement of quantities and identities of materials in a mixture. This method is broadly used in biomedical research, as well as in the creation, manufacture and control of therapeutic interventions. The current proposal is to use new knowledge in materials science and chemistry to enable faster and more efficient separations by liquid chromatography, saving time and money, as well as enabling new uses of the method to understand the structure and function of biological molecules.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Small Business Innovation Research Grants (SBIR) - Phase I (R43)
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Special Emphasis Panel (ZRG1-IMST-G (10))
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Edmonds, Charles G
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Advanced Materials Technology, Inc.
United States
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DeStefano, Joseph J; Boyes, Barry E; Schuster, Stephanie A et al. (2014) Are sub-2 ?m particles best for separating small molecules? An alternative. J Chromatogr A 1368:163-72
Destefano, Joseph J; Schuster, Stephanie A; Lawhorn, Jason M et al. (2012) Performance characteristics of new superficially porous particles. J Chromatogr A 1258:76-83
Wagner, Brian M; Schuster, Stephanie A; Boyes, Barry E et al. (2012) Superficially porous silica particles with wide pores for biomacromolecular separations. J Chromatogr A 1264:22-30