High performance liquid chromatography (HPLC) uses high pressure to force solvents through a variety of columns which have extremely small particles that allow for extremely high resolution separation. HPLC is a valuable resource in many settings such as: pharmaceutical, food, and cosmetics. The use of HPLC in food chromatography over the past fifty years has risen tremendously. Learning HPLC in an undergraduate program is beneficial for students as it offers them experience for industry. Other undergraduate students have developed labs using HPLC, such as to analyze Vitamin C in food. Recently, Lock Haven University received two Waters Alliance 2695 HPLC’s with photo diode array (PDA) detectors, with a temperature controller for the column, and Empower 3 software. Methods need to be developed to facilitate the use of these advanced instruments in the undergraduate teaching laboratory. Methods are needed for analysis and automated report generation so that large groups of samples can be analyzed automatically using the instruments auto sampler and report-generating software. In addition, methods for advanced analysis techniques such as analysis of 3D plots and wavelength selection need to be developed for research an other advance work. A document was developed detailing sample analysis and report generation for others to be able to use the instrument. Initially, a trial and error technique was used with the software to learn some of the capabilities. Through research, some Waters documentation and chromatography forums were found to assist in the understanding of the materials. Further work can be done on the instrument and software to be able to do more as the Empower 3 software has many abilities in analyzing and processing data.

Development of high performance liquid chromatography methods for a Waters Alliance 2695 running Empower 3 software

253rd National Meeting (2017)

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