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Fructose is one of the most important monosaccharides in the human diet that the human body needs for proper metabolism. This paper presents an approach to study biochemical changes caused by sugars in human normal bronchial cells (BEpiC) and human cancer lung cells (A549) by Raman spectroscopy and Raman imaging. Results after supplementation of human bronchial and lung cells with fructose are also discussed and compared with results obtained for pure human bronchial and lung cells. Based on Raman techniques we have proved that peaks at 750 cm−1, 1126 cm−1, 1444 cm−1, 1584 cm−1 and 2845 cm−1 can be treated as biomarkers to monitor fructose changes in cells. Results for fructose have been compared with results for glucose. Raman analysis of the bands at 750 cm−1, 1126 cm−1, 1584 cm−1 and 2845 cm−1 for pure BEpiC and A549 cells and BEpiC and A549 after supplementation with fructose and glucose are higher after supplementation with fructose in comparison to glucose. The obtained results shed light on the uninvestigated influence of glucose and fructose on lipid droplet metabolism by Raman spectroscopy methods. [Display omitted] • Raman spectroscopy allows to identify the biomarkers in BEpiC and A549 cells caused by incubation with fructose and glucose. • Raman bands at 750 cm-1, 1126 cm-1, 1444 cm-1, 1584 cm-1 and 2845 cm-1 can be treated as biomarkers to monitor fructose changes in BEpiC and A549 cells. • Raman spectroscopy allows to monitor the metabolism of lipid droplets after supplementation with glucose and fructose. [ABSTRACT FROM AUTHOR]