Copyright © 2006 Elsevier B.V. All rights reserved.
Short communication
Thermal stability of glucose and other sugar aldoses in normal phase high performance liquid chromatography
Received 10 March 2006;
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Abstract
Analysis of glucose and other simple sugars are often performed by use of normal phase HPLC methods with acetonitrile as major eluent. The present results clearly show that column temperature plays an important role with respect to chromatographic performance and detection limits of glucose when using a specific carbohydrate column. A change in column temperature from 25 to 45 °C reduced the detection of glucose (with ELSD) by more than 41%, whereas the detection of other sugar aldoses (galactose, xylose and rhamnose) were suppressed even more. By increase of column temperature to 70 °C the detector signal of glucose was found to be less than 2% compared to that obtained at 20 °C. Neither fructose nor sucrose showed similar correlation between column temperature and detection. The rate of decreased response is not dependent on sample concentration or the ELSD settings. The results express the importance of accurate temperature control in the analysis of sugar aldoses, and also values low column temperatures for samples with low concentrations of sugar aldoses in order to improve detection.
Keywords: Aldoses; Glucose; Ketoses; Fructose; Sugars; HPLC; Normal phase; Column temperature; Evaporative light scattering detection (ELSD)







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0.070 μg/mL) can be obtained so the suppressed eluent could directly go into an ELSD detector without obvious interference of inorganic salts. After examining the changes in retention and resolution, an optimized method was established (for IC: using 32 mM KOH as the eluent at a flow rate of 1 mL/min; for ELSD: operated at 95 °C, 4.0 bar nitrogen with a gas flow rate of 2.0 L/min) and the linearity, reproducibility, and the limit of detection (LOD) for the three carbohydrates were further evaluated. Regression equations revealed acceptable linearity (correlation coefficients = 0.994–0.998) across the working-standard range (100–1000 μg/mL for glucose and sucrose, 150–1000 μg/mL for fructose) and LODs of glucose, fructose, and sucrose were 93, 126, and 90 μg/mL, respectively. This method has successfully been applied to the determination of the three carbohydrates in carbonated cola drinks and fruit juices. The recoveries were between 95 and 113% (





