Forth-eight cup-shaped edible birds nest (EBN) harvesting from natural cave and man-made house were collected from Malaysia and Indonesia to study the distribution characteristics and traceability of 27 elements in EBN. The inductively coupled plasma mass spectrometry (ICP-MS) was applied for the detection of multi-elements in EBN, and five macro-elements which distributed according to the rule of Na＞Ca＞Mg＞K＞P were found in EBN. Besides, in EBN, some essential trace elements (Fe, Cu, Zn, Mn, Mo, Co, Se, Cr), 3 light rare earth elements (LREE) and 2 heavy rare earth elements (HREE) also be detected. All of 27 elements in EBN is skewed distribution with bigger coefficient of variation (COV) except for Pt. The t-test, Mann-Whitney U, Rank Cases based on the Tukey formula and Double factor variance analysis was used to analyze the influence of geographical origin, harvesting method and their interactions on the distribution of multi-elements in EBN. B, Na, P, Ca, Mn, Cu, Sr, Mo, Ba and Nd were significantly relative with harvesting method of EBN, and Mg, Al and Pt were for geographical origin of EBN. This article not only verified the feasibility to trace the provenance of cup-shaped EBN, but also provided a reference for traceability research, Nutritional function development and quality grade assessment of EBN.