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A method for identification of foreign amylases in honey
Erban, Tomáš ; Shcherbachenko, Elena ; Talacko, Pavel ; Harant, Karel
Honey is a unique natural product. Honey has been used as a sweet and delicious foodstuff since ancient times. However, it is also valued for its multifaceted currative properties. Unfortunately, honey is one of the most adulterated foods. Nothing may be added to or modified from the honey. Honey also needs to be handled with care. Honey quality can negatively be affected by the way of processing such as heating and storage. Despite great progress in analytical methods, it is not possible to prove all adulterated honeys. Some methods of adulteration are quite sophisticated. Therefore, it is necessary find new approaches and methods for identification of honey adulteration. To be sold, honey must comply with internationally valid standards, which are also valid at national levels with possible minor modifications. One of the important parameters for honey is the level of diastase or amylase enzyme activity, which is a recognized indicator of the freshness and quality of honey. Lower diastase activity below the established level may indicate old honey, but it may also be the result of careless handling of honey. Last but not least, diastase activity may be reduced due to adulteration of honey such as its dilution with sugar substitutes. It is possible that amylase activity can be artificially adjusted by the addition of enzymes. Honey adulterated by the artificially added amylase meets the legislative requirements for placing honey on the market, but this violates the rules laid down by law. The methods used so far have not made it possible to prove this way of honey adulteration. Therefore, this methodology focuses on the identification of foreign amylases that may occur in honey. The methodology enables to identify practically any foreign amylase in honey by bottom-up shotgun proteomic approach. Based on the obtained results with specific peptides can be further used for the development of a targeted method for the identification of foreign amylases.
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