The propolis used in this study is propolis from Brunei Darussalam, including G. thoracica, H. itama, and T. binghami. This study focuses on evaluating phytochemicals, including the total content of polyphenols and flavonoids, marker compounds, and the anti-fungal activity of propolis Brunei. Until now, research on the compounds contained in propolis is still being carried out. However, the literature on the chemical compound of Brunei propolis is still limited. The results of the research on the content of Brunei propolis using LC-MS/MS found as many as 21 chemical compounds and three marker compounds, namely maslinic acid, D-(-) Mannitol, and 18-β-Glycyrrhetinic acid. The total content of flavonoid and polyphenolic propolis in Brunei was obtained using quercetin as a flavonoid standard and gallic acid as a polyphenol standard. In Brunei propolis, the total flavonoid content was higher than the total polyphenol content. Where the total polyphenol content of propolis G. thoracica, H. itama, and T. binghami were 78.79±17.06; 70.51±12.93 and 16.37±0.53 (mgGAE/g). while the total flavonoid content was 19.30±1.99; 101.10±6.26 and 61.63±4.53 (mg QE/g). The antifungal activity was carried out by agar diffusion and microdilution methods. Brunei propolis extract showed antifungal activity against Cryptococcus and C. Albicans, whereas propolis Brunei extract showed anti-fungal activity with intermediate resistance to both fungi.