A validated method for the quantitative determination of sugars in honey using high-performance thin-layer chromatograph
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IGINAL RESEARCH PAPER
A validated method for the quantitative determination of sugars in honey using high‑performance thin‑layer chromatography Md Khairul Islam1,2 · Tomislav Sostaric2 · Lee Yong Lim2 · Katherine Hammer1,3 · Cornelia Locher1,2 Received: 11 March 2020 / Accepted: 8 September 2020 © Akadémiai Kiadó, Budapest, Hungary 2020
Abstract Sugars, in particular glucose and fructose, are the main constituents of honey, comprising up to 85% of its total weight. A high-performance thin-layer chromatography (HPTLC) method to identify and quantify common sugars (glucose, fructose and sucrose) in honey has been developed and fully validated according to the International Conference on Harmonisation guidelines. It allows to determine a honey’s fructose-to-glucose ratio, which is not only an important authentication and quality control parameter, but also an indication of its tendency to crystallise. The HPTLC analysis is easy to perform, accurate, precise, specific and sensitive and requires only minimal sample preparation. With a limit of detection/limit of quantification of 21.98 ng/66.62 ng for fructose, 33.00 ng/100.00 ng for glucose and 21.17 ng/64.15 ng for sucrose, the sensitivity of the method has been greatly improved compared to other HPTLC-based approaches. An additional advantage is the method’s simplicity and fast processing time as it only requires a single development step without plate or sample pre-treatment. Keywords High-performance thin-layer chromatography (HPTLC) · Sugar · Quantitative analysis · Honey
1 Introduction Sugars play an important role in many natural products and food items. Honey, for example, is a highly concentrated sugar solution, its main sugars being fructose and glucose, which account for about 80–85% of the total solid. They are also in the main responsible for the sweetness of honey, with fructose being twice as sweet as glucose [1]. Other sugars present in honey include disaccharides (e.g., sucrose, maltose, trehalose, turanose), trisaccharides (e.g., maltotriose, raffinose, erlose) and oligosaccharides [2]. As fructose and glucose are the major constituents of honey, their identification and quantification is of great interest. In particular, the fructose-to-glucose ratio is important to the beekeeping industry as it contributes to the authentication of a honey’s floral source, assists with the identification * Cornelia Locher [email protected] 1
Cooperative Research Centre for Honey Bee Products Limited (CRC HBP), Perth, Australia
2
Division of Pharmacy, School of Allied Health, University of Western Australia, Crawley, WA 6009, Australia
3
School of Biomedical Sciences, University of Western Australia, Crawley, WA 6009, Australia
of honey adulterants (e.g., feeding of bees with sugar syrups during times of low nectar flow or adding sugar syrups to honey to increase honey yields [3–8]) and also allows to predict the typical crystallisation behaviour of honey [9–12]. A honey’s sugar composition can be determined by several analytical methods, incl
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