Comparison of ultraviolet and refractive index detections in the HPLC analysis of sugars

Abstract Refractive index (RI) detection is the standard approach for quantitatively detecting sugars via high-performance liquid chromatography (HPLC), while ultraviolet (UV) absorbance detection is the most commonly used detection method for general HPLC analysis. We compared the two detection approaches of UV and RI in the HPLC analysis of small sugars to investigate whether UV detection could be an alternative method to RI detection. UV detection was performed using a photodiode array scanning from 190 to 400 nm. We obtained comparable limit of detection (LOD) results for RI and UV detection in the HPLC analysis of monosaccharides, while HPLC-RI provided better LOD results than HPLC-UV in disaccharide analysis. Both HPLC-RI and HPLC-UV methods were applied to analyze a real honey sample, and similar results were obtained in terms of precision and recovery. The study conclusively shows that the UV-based HPLC analysis of sugars offers a sufficient alternative to RI-based HPLC analysis.