*2.1. Materials*

Animal ghee was purchased from Unilever Holdings Sdn. Bhd. (Kuala Lumpur, Malaysia). Calcium and vitamin D were purchased from Eurobio Sdn. Bhd. (Victoria, Australia). Orlistat was purchased from Xepa-Soul Pattinson Sdn. Bhd. (Melaka, Malaysia), cholesterol powder from Nacalai Tesque (Kyoto, Japan), Eosin Y from Sigma-Aldrich (St. Louis, MI, USA), haematoxylin from Richard-Allan Scientific (Kalamazoo, MN, USA), and oil-Red O from VWR Life-Science AMRESCO (Solon, OH, USA). All other reagents were of analytical grade.

#### *2.2. Preparation of Bee Bread Sample*

Bee bread from stingless bee (*Heterotrigona itama*) was purchased from local stingless bee farm from Selangor, Malaysia. The sample was dried using food dehydrator at 35 ◦C. Then, it was ground into fine powder using a mini blender and kept in a sterilised container at –20 ◦C until further analysis.

#### *2.3. Liquid-Chromatography-Mass Spectrometry Analysis of Bee Bread*

The presence of phenolic compounds was assessed by liquid chromatography-mass spectrometry. Based on studies conducted by Isidorov et al. [10] and Urcan et al. [21], bee bread was screened for the presence of fourteen phenolic compounds, which included apigenin, benzaldehyde, ca ffeic acid, chrysin, ferulic acid, gallic acid, hydroquinone, isorhamnetin, kaempferol, mangiferin, naringenin, p-coumaric acid, quercetin, and resveratrol. Briefly, ten gram of powdered bee bread was soaked in 5 mL of methanol and sonicated (Lab Companion, Model UC-20, Seoul, Korea) for 30 min. The solution was filtered and evaporated using rotovap (Buchi, Rotavapor ® R-300 system, Flawil, Switzerland) to make a stock solution of 3 mg/mL. The resulting solution was filtered through a membrane filter (pore size 0.22 μm) before analysis. The sample was analysed by LTQ Orbitrap mass spectrometer (Thermo Scientific, San Jose, CA, USA). Acetonitrile and 0.1% formic acid were used as the mobile phase. The spectral *m*/*z* from 100–1000 was recorded in positive ionization mode. The mass spectrophotometry was performed in electrospray ionisation conditions and positive mode with the following parameter settings: source accelerating voltage = 4.0 kV; capillary temperature = 280 ◦C; sheath gas flow = 40 arb; auxiliary gas = 20 arb.
