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Lab #9: Basics of High Performance Liquid Chromatography (HPLC): Determination of Toluene and Anthracene in an Unknown Solution Tiffany Lam, Sam Kim Department of Chemistry, Binghamton University, Binghamton, NY 13902
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Abstract The objective of the lab was to determine the elution order of individual components in mixture, understand the methods of quantization in chromatography, and measure the fundamental parameters of chromatography. The HPLC technique was applied to measure the amounts of toluene and anthracene in unknown solution. This was accomplished by setting up five standard samples containing different amounts of toluene, anthracene and internal standard m-xylene. Area of the peaks produced by the chromatograms were utilized to calculate the response ratio. The response ratios were plotted against %v/v of toluene and anthracene, which was then used to calculate the unknown amounts of each compound. Toluene was 45. 601%v/v and anthracene was 2.282%v/v of the unknown solution. Introduction High performance liquid chromatography (HPLC) is an advanced form of column chromatography. Instead of a solvent dripping through a column by force of gravity, it is forced through under high pressures making the process much faster. The columns in HPLC are made of smaller sized particles which gives greater surface area for interactions between stationary phase and molecules travelling in the mobile phase. This allows better separation of individual components in the complex mixture. Another improvement is the highly automated and sensitive detection methods. Reversed phase HPLC is the most commonly utilized form of high performance liquid chromatography. The solid support or column is made of long, nonpolar hydrocarbon chains. The mobile phase is a polar solvent such as a mixture of water and alcohol. Polar molecules in mixture spend most of the time moving with the mobile phase because of strong attractions between same polarity. Nonpolar molecules in mixture tend to be attracted to the hydrocarbon
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surface because of van der Waals forces therefore they spend less time in solution with solvent. In other words, polar molecules travel faster through the column than nonpolar molecules.
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  • Spring '14
  • Chuan-JianZhong
  • Chromatography, High performance liquid chromatography

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