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.
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