Abstract
Five HPLC methods were employed for the quantitative analysis of three natural sugars namely fructose, glucose and sucrose in soft drinks. HPLC-refractive index detector (RID)-AMINO proved to be the most suitable HPLC method to carry out the latter task. For the optimum separation and response of the natural sugars the best conditions employed were column oven temperature 30 oC, flow rate 0.1 mL/min, mobile phase ratio acetonitrile:water 75:25 and they were determined by studying all possible interactions among these three parameters. Full validation of HPLC-RID-AMINO was performed in terms of system suitability test, precision check, accuracy check and robustness.
 
 KEY WORDS: Sugar, Soft drink, Experimental design, Validation, HPLC, System suitability
 
 Bull. Chem. Soc. Ethiop. 2020, 34(2), 419-426
 DOI: https://dx.doi.org/10.4314/bcse.v34i2.17
Highlights
Diabetic patients are advised to limit their sugar intake [1, 2] in compliance with the studies showing that simple sugars can cause higher postprandial glycemia than starch [3]
The work presented in this paper focuses on how to come up with the best suited High performance liquid chromatography (HPLC) method with respect to system suitability, method performance, volume of sample used, organic wastes generated and cost and on how to determine its optimum conditions that would allow simultaneous separation, detection, identification and quantification of the three natural sugars present in a sample of a soft drink
HPLC-refractive index detector (RID)-AMINO (Figure 1) was considered the best among all the HPLC methods employed in this work since it used lesser mobile phase (1.8 mL of Acetonitrile per sample) as compared to the HPLC-RIDHILIC and HPLC-RID-ANIONIC where 6 mL of acetonitrile per sample were used
Summary
Diabetic patients are advised to limit their sugar intake [1, 2] in compliance with the studies showing that simple sugars can cause higher postprandial glycemia than starch [3]. To avoid problems such as stimulating hyperglycemia, having recourse to insulin [4, 5], and causing possible cardiomyocyte dysfunction [6, 7] and/or enhanced loss of β-cells [8] diabetic patients have diets low in sugar. Amongst Mauritian adults aged 45 years and over, approximately 1 in 2 people have diabetes, or have a high risk of developing it [9]. High performance liquid chromatography (HPLC) is the most adopted technique for the separation and quantification of individual natural sugars namely glucose, fructose and sucrose [10]. Non-structural carbohydrates may be isolated and quantified using HPLC coupled with the relevant columns and detectors such as refractive index detector (RID), evaporative light scattering detector (ELSD) and pulsed amperometric detector (PAD) [10,11,12]
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