Hyaluronic acid is a naturally occurring linear polysaccharide with substantial medical potential. In this work, discrimination of tyramine-based hyaluronan derivatives was accessed by ion mobility-mass spectrometry of deprotonated molecules and nuclear magnetic resonance spectroscopy. As the product ion mass spectra did not allow for direct isomer discrimination in mixture, the reductive labeling of oligosaccharides as well as stable isotope labeling was performed. The ion mobility separation of parent ions together with the characteristic fragmentation for reduced isomers providing unique product ions allowed us to identify isomers present in a mixture and determine their mutual isomeric ratio. The determination used simple recalculation of arrival time distribution areas of unique ions to areas of deprotonated molecules. Mass spectrometry data were confirmed by nuclear magnetic resonance spectroscopy.
Increased use of food, beverages or drugs containing synthetic sweeteners presents a real danger to health, which is why the EU Member States had to establish a system of regular surveys to monitor sweetener consumption. In case of wine industry, according to law no. 244/2002, Romania prohibits the addition of synthetic sweeteners in wine in order to obtain sweet wine. The official method for detection of adulterated sweet wines with synthetic sweeteners is TLC-Thin Layer Chromatography. However, quantitative methods of analysis are needed to measure levels of sweeteners in different food matrices and high performance liquid chromatography has proved to be a powerful tool for quantitative analysis of compounds at traces levels. In this paper, a high performance liquid chromatographic (HPLC) method for the simultaneous separation and determination of three of the most popular artificial sweeteners (acesulfame potassium, saccharine and aspartame) in a single injection was developed. The described method is rapid, accurate and highly sensitive. Detection limit were 4 mg/L for acesulfame K, 1 mg/L for saccharine and 9 mg/L for aspartame respectively. The precision of the method was about 2% and recovery ranged between 92.6% and 103.3%. There were analyzed commercial wine samples, in order to detect possible counterfeits-sweet and medium sweet wines. Therefore, of the 20 analyzed wine samples, only two samples consisting in wine sweet table, were counterfeited by adding saccharin.
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