*2.3. EBN Extraction*

EBN extraction was performed according to a previous study [28], with some modifications. Water extraction was selected to simulate the traditional technique commonly used before the consumption of EBNs. Water extraction gives a general idea of the protein consumed by people. EBN extraction was done using four types of methods: stew (SE), full stew (FS), sonication (SO), and hot water (HW). The cleaned EBN was ground and soaked with deionized water, with a ratio of 1:100 (*w/v*), for 16 h at 4 ◦C. All the extracts were subsequently frozen at −80 ◦C, lyophilized, and stored at −20 ◦C until further use.

#### 2.3.1. Stew and Full Stew Extraction

SE and FS extraction methods were carried out using a double-boiling technique, where the vapor from the boiling water in the bottom pot heated the top pot containing the EBN. The soaked EBN was double-boiled for 30 min at 100 ◦C and cooled to room temperature. The SE extract consisted of the double-boiled EBN, filtered using a muslin cloth, and the FS extract consisted of the whole double-boiled EBN, without filtration.
