Separation and Determination of d -Allose in Presence of Process-Related Impurities by Capillary Electrophoresis
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Separation and Determination of D-Allose in Presence of Process-Related Impurities by Capillary Electrophoresis Sri Rama Krishna Surapureddi 1,2 & Kunta Ravindhranath 1
2
& Ghantasala S. Sameer Kumar & Sreedhar Reddy Sappidi
2
Received: 23 April 2020 / Accepted: 25 August 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract A simple and effective capillary electrophoresis (CE) method for the separation and quantification of D-allose in the presence of its process impurities, namely sucrose, D-glucose, D-fructose, D-psicose, and D-altrose, has been developed and validated. The optimum conditions for the separation with an acceptable resolution are found to be as follows: background electrolyte (BGE): 36 mM of Na2HPO4 and 130 mM NaOH; pH: 12.6; and voltage: 18 kV with direct UV detection at 265 nm. The present method overcomes the limitations of existing analytical method like sample preparation, poor resolution, and sensitivity. The method is optimized with the established standard curves for D-allose and its related impurities in the concentration range 0.25 to 3.0 mM with coefficient of determinations > 0.99 and the limit of detection (LOD) in the range of 0.11 to 0.20 mM. Separation of D-allose and its impurities present in the mixture is achieved using the current method with good resolution, linearity, and reproducibility. The present method is suitable for the analysis of the final product in the enzymatically synthesized D-allose and also for reaction monitoring of in-process samples. Keywords Carbohydrate analysis using capillary electrophoresis . D-Allose with processed impurities . D-Psicose . Enzymatic conversion of rare sugars . D-Altrose
Introduction D-Allose, C-3 epimer of glucose, is a rare sugar and it exists in
trace amounts in various natural sources (Perold et al. 1973; Jensen and Nielsen, 1981; Chari et al. 1981; Weckwerth et al. 2004; O’Neil et al. 2006; Ragupathi Raja Kannan et al. 2012; Sithara et al. 2017). This monosaccharide has numerous physiological and medicinal benefits over common sugars. By possessing 80% sweetness of table sugar and with ultra-low calorific value, D-allose becomes a promising food additive or supplement to prevent excessive weight gain (Chen et al. 2018). Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12161-020-01842-z) contains supplementary material, which is available to authorized users. * Kunta Ravindhranath [email protected] 1
Department of Chemistry, Koneru Lakshmaiah Education Foundation, Green Fields, Vaddeswaram, Guntur District, Andhra Pradesh 522502, India
2
Vimta Labs Ltd, Genome Valley, Hyderabad, Telangana 500085, India
Due to the scarcity of D-allose, researchers are synthesizing it through chemical, enzymatic (Chattopadhyay et al. 2014; Li et al. 2013), and microbial (Leang et al. 2004) routes. Chemically synthesized products have several disadvantages such as low yield, unwanted impurities, and chemical contamination. So, various enzymatic pathways a
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