Determination and Standardization of a Method for Caffeine in the Roasting Process Chromatographic High Performance Liquid (Hplc)

The work done in the organization. COFFEE COLONIAL.SAS was to evaluate, analyze and verify key information for the identification and standardization of a method for caffeine in coffee roasting process by the technique of chromatography high performance liquid (HPLC).Assays were performed at different roasting process to get the right conditions for the samples, supported by transport phenomena such as, energy, fluids and heat, in addition to following the guidelines of the Standard ISO 2859-1 Colombian NTC.To obtain a standard curve, leading to interpolate coffee samples, prepare a solution with Caffeine (Sigma-Aldrich, Analytical Reagent) and Milli-Q water (Millipore Merck 40 ppm), to obtain a solution pattern, from the solution of caffeine more solutions were prepared by 5 micrograms per milliliter caffeine.To determine the most suitable mobile phase, it began testing different samples of caffeine of coffee with the same water-acetonitrile mobile phase in different proportions, once all the necessary tests and according to the results obtained, the mobile phase allow better separation and interaction with the stationary phase was composed of water-acetonitrile mobile phase in 20:80 ratio.After obtaining the calibration curve and the more appropriate mobile phase, where injections were made different chromatograms were obtained with bases and defined peaks, which are defined by giving a correlation coefficient of linearity close to one, after which the next step was to find the area under the curve to define the caffeine concentration of the samples and thus being able to express in percentage.