Abstract: House edible bird's nest and cave edible bird's nest are two main types of EBN. However, the key differential components, which can cause the differences of these EBN are not clear. In order to explore the differences between house EBN and cave EBN and effectively discriminate between different EBN samples, protein, amino acid, sialic acid, mineral elements and other nutritional components, as well as the hazard factors such as nitrite and heavy metal elements in four kinds of house EBN and cave EBN were determined, respectively. Moreover, cluster analysis and key differential components screening were carried out using heatmap and partial least squares regression (PLSR), respectively. The results showed that protein and sialic acid were the most important nutritional components, accounting for 63.09%~67.79% and 7.22%~10.41% in the total EBN, respectively. Leucine, valine, threonine and phenylalanine were the main essential amino acids in EBN, while serine, aspartic acid and proline were the main non-essential amino acids. In addition, the proportion of essential amino acids in total amino acids (45.76%~47.60%) was higher than 40% of the standard according to the high-quality protein recommendation from FAO/WHO. The contents of protein (66.70~67.79 g/100 g>63.09~66.28 g/100 g) and sialic acid (10.41 g/100 g<7.22~7.88 g/100 g) in house EBN were higher than those in cave EBN. The contents of amino acids and mineral elements in cave EBN were significantly higher than those in house EBN (P<0.05). On the other hand, house EBN and cave EBN could be well distinguished by PLSR analysis. Trace elements, such as Se, Fe, I, Mn, Cu, not only were the key differential components between the two EBN, but could be used as the basis to discriminate the origin of EBN species. Cave EBN has an advantage in the content of basic nutrients, and the detection of trace elements can serve as a basis for distinguishing different EBN samples.