This study describes the approach of amino acid and monosaccharide combined with Hotelling T2 range plot to identify edible bird nests (EBN) and non-EBN. Prior to the approach, an analytical method was developed and validated to quantify monosaccharides in EBN. Hotelling T2 range plots of both compounds were successful in predicting the different types of EBN and differentiating EBN and non-EBN. This outcome suggests EBN contains a group of glycoproteins which is not affected by the EBN’s coloration, country of origin, and/or the processing method of the food item. In addition, the glycoproteins were shown to be unique to EBN. EBN were revealed to be rich in protein and essential amino acids as well as contain a wider variety of monosaccharides than most food items. The overall findings suggest that amino acid and monosaccharide provide information not only on the detected compounds and also insights into the glycoproteins of EBN.