UHPLC : Perspectives, Performance, Practices, and Potential Issues

Ultra-high-pressure liquid chromatography (UHPLC) is a modern high-pressure liquid chromatography platform offering higher performance in speed, resolution, and precision. It is useful in method development and suitable for regulatory analysis. This chapter provides an overview of UHPLC systems and applications including historical perspectives, fundamental concepts, benefits, potential issues, and best practice. It first provides a brief review of chromatography concepts that are essential for a better understanding of UHPLC and its applications. The chapter then summarizes the benefits of UHPLC including fast separations, high-resolution analysis, rapid method development, flexibility for customizing resolution, and other performance enhancements. Each benefit is described with case studies. The use of UHPLC under high operating pressures with low-dispersion equipment can present potential issues for many new users. Earlier issues have been mitigated with improved instrumentation and the introduction of second-generation UHPLC equipment. Each potential issue is described here along with its mitigation strategies.