Abstract:The adulteration of the honey industry is serious, especially the syrup adulteration method is difficult to detect, and a convenient and economical honey detection technology is needed. In this paper, solid phase extraction was used to enrich oligosaccharides with a polymerization degree of more than five, and thin-layer chromatography image analysis was used to detect the incorporation of fructose syrup in honey. The enrichment effects of different solid phase extraction conditions on oligosaccharides were compared. The effects of different developing system on the separation of oligosaccharides were investigated. The results showed that there was no oligosaccharide with a degree of polymerization of more than five in natural honey, and honey mixed with fructose syrup could detect oligosaccharides of more than five sugars. The image of the malt heptasaccharide in adulterated honey was analyzed by Image J and Sorbfil software, and the standard curve was drawn. The content of maltose in the sample was determined by external standard method. The linear relationship of maltoheptaose in the range of 1.0~5.0 mg/mL was good, and the correlation coefficients were 0.9946 and 0.9948, respectively. When the level of fructose syrup was 20%~40%, the average recoveries obtained by the two softwares were 98.56% and 101.20%, respectively. The detection limits obtained by the two softwares were 0.037 mg/mL and 0.061 mg/mL,respectively, and the limit of quantification was 0.11 mg/mL and 0.19 mg/mL, respectively. The method is sensitive and accurate, and relies on software operation. The thin-layer chromatogram image can be stored for a long time, requires no expensive instruments, is economical and simple, and is suitable for the detection and quantification of fructose syrup in honey.