.2. Determination of caffeine in coffee beans
A UV/vis spectrophotometer method cannot be used directly for the determination of caffeine in coffee seeds owing to the matrix effect of UV absorbing substances in the simple matrix (Ortega-Burrales et al., 2002 and Zhang et al., 2005). This effect is also clearly seen in the spectral bands of caffeine in coffee seeds (Fig. 5) dissolved in water. Hence, it is not suitable to determine the percentage of caffeine in coffee seeds due to overlapping of these interfering bands. In order to overcome this difficulty coffee was first dissolved in water and caffeine extracted from solution using dichloromethane. Dichloromethane is the most commonly employed method for extraction of caffeine from green coffee beans (Rofti, 1971). Many commercial products applied dichloromethane for decaffeinating the coffee beans for its extraction efficiency 98–99% (Clarke, 1980). The extraction was made four times until the spectrum of caffeine becomes flat when seen under UV/vis spectrophotometer. Fig. 6 below shows peak absorbance versus wavelength of caffeine spectra extracted using dichloromethane from water solution at different stages. For comparison the spectra of each stage were overlapped. It is clearly shown in Fig. 6 that the concentration of caffeine at each stage is different, hence the size of peak absorbance also varies. From the results obtained high amount of caffeine concentration was extracted in the first stage of extraction. Moreover as seen from the graph almost no caffeine peak was seen in the four rounds of extraction.