Establishment of a holistic and scientific protocol for the authentication and quality assurance of edible bird’s nest

• Authentication system of edible bird’s nest (EBN) was established by three methods.
• Epidermal growth factor (EGF) was determined by immunoblotting technique.
• Oligosaccharide fingerprints were confirmed by gas chromatography–mass spectrometry.
• Surface morphology was observed by environmental scanning electron microscopy.
• Genuine EBN presences five monoses, EGF and unique network microstructure.

Edible bird’s nest (EBN) is a prestigious and superior functional food. It is expensive due to its limited supply and enormous demand. Consequently, many fake products are available in the market. This report aims to design a holistic and scientific testing method for the authentication and quality assurance of EBN. The analytical system involves a concerted approach by applying the gas chromatography–mass spectrometry (GC–MS) fingerprint of oligosaccharides, the environmental scanning electron microscopy (ESEM) of microstructure, and the immunoblotting of epidermal growth factor (EGF) in EBN. The results confirmed that genuine EBN had the presence of five monoses and EGF, whereas the counterfeit EBN did not. Moreover, the content of N-Acetylneuraminic acid (NANA) and EGF are established as unique indicators for the grades of EBN. A unique three-dimensional, crater-like microstructure was also observed in authentic EBN, but not in fake products. It is concluded that the holistic approach, including chemical, physical and biochemical studies of EBN, is a reliable and scientific method for the verification of EBN.