In my thesis, I identified some artificial sweeteners in soft drinks. First, I optimized the separation on the HPLC instrument. In various articles, I have found methods for simultaneous determination of sweeteners. I decided to determine acesulfame K and saccharin. I used reverse non-polar stationary phase C$_8$ and polar mobile phase with gradient separation of acetonitrile and 0,5 % phosphoric(V) acid. The injection volume was 10 µL and a constant flow rate of 0,700 mL/min. I set the wavelenght of the detector to 220 nm so that I could determine acesulfame K and saccharin. I made a calibration curve in the concentration range between 1-50 mg/L for both standards. I set the Limit of Detection and Limit of Quantitation. The Limit of Detection was 0,1 mg/L and the Limit of Quantitation was 0,3 mg/L. Then I determined the cocentrations of acesulfame K and saccharin in four samples of soft drinks. Acesulfame K was present in all samples, while saccharin was detected in only one sample. The method was appropriate, as I had the repeatability of the samples were comparable to each other. All RSD values were below 5 %. The concentrations of acesulfame K and saccharin in the samples were very low and below the acceptable daily intake. During the experimental work, I spotted 3 chromatographic peaks that I wanted to identify. I looked at the declaration for possible compounds that could belong to these chromatographic peaks: ascorbic acid, citric acid, phenylalanine, malic acid, potassium sorbate and caffeine. Of these compounds, I proved two. These were potassium sorbate and caffeine.
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