Identified Compounds from Ethyl Acetate Phase of Temu Mangga (Curcuma mangga Val.) Using LC-MS/MS and Their Potential as Anticancer Against MCF-7 Cells

Curcuma mangga Val. is a well-known species of the Zingiberaceae family. In previous research, acetone extract of this species was highly active in inhibiting MCF-7 cells’ proliferation (IC₅₀ = 1.45 μg/mL), where crude acetone extract was partitioned consecutively using n-hexane, ethyl acetate (EtOAc), and methanol. The soluble components in the n-hexane phase, identified as zedoarondiol, curcumenol, curcumenone, and 13-hydroxygermacrone, are thought to inhibit MCF-7 cells’ proliferation. In contrast, the soluble compounds in the EtOAc phase have not yet been reported. Therefore, this study aims to identify the soluble compounds in the EtOAc phase of acetone extract and examine their anticancer activity toward MCF-7 cells. Fractionation and purification of the EtOAc constituents are achieved by vacuum liquid chromatography, radial chromatography, and preparative thin-layer chromatography, and the chemical structure of the isolated compounds is identified by employing liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Furthermore, the isolated secondary metabolites are examined against the MCF-7 cell line using a 3-(4,5-dimethylthiazol-2-il)-2,5-diphenyltetrazolium bromide (MTT) assay. By following these protocols, three fractions are obtained. LC-MS/MS spectra show that fraction 1 (α) contains an unidentified flavanone or chalcone compound, fraction 2 (β) confines zedoarofuran, and fraction 3 (γ) is a mixture of zederone, curzerenone, and zedoarofurane. Meanwhile, fractions 2 (β) and 3 (γ) are terpenoid compounds. The MTT assay shows that fraction 2 (β) contains zedoarofuran (eudesmane-type sesquiterpenoids) and is moderate at inhibiting MCF-7 cells’ proliferation (IC50 < 31.25 µg/mL). Fractions 1 and 3 are inactive with IC₅₀ < 50 µg/mL.