## 3 Results
### Characterization and Structural Elucidation of the Isolated Lupeol  
Characterizarion of the isolated white amorphous showed that it has a melting point of 216 \({}^{\circ}\)C. Meanwhile, the IR spectrum of the isolated compound was also studied (Supplemental Figure S2). The hydroxyl group (-O-H) vibration was shown as a broad intense band at 3303 cm\({}^{-1}\), and a moderately intense band was observed at 1187 cm\({}^{-1}\). The stretching and bending of the methyl group (-C-H\({}_{3}\)) were detected at 2930 cm\({}^{-1}\) with an intense band and a moderately intense band at 1450 cm\({}^{-1}\). In addition, the vibration of the (-C-H\({}_{2}\)) methylenic part was observed at 2861 cm\({}^{-1}\), which displayed a weaker intense band. The vibration of the (-C=O) of the carboxylic acid group was also observed as an intense band at 1737 cm\({}^{-1}\). At the wavelength of 1379 cm\({}^{-1}\), there was the presence of (-C-C) vibration of the methyl group as a moderate intense band. The out of the plane (-C-H) vibration was observed at 886 cm\({}^{-1}\) as a strong intense band. Those obtained IR spectra data were in good agreement with the IR spectra of lupeol, as studied and reported by [12; 13]. Elucidation of the structure of the isolated compound was performed using \({}^{1}\)H-NMR (600 MHz) and \({}^{13}\)C-NMR (150 MHz). The \({}^{1}\)H-NMR spectrum of the compound showed the presence of seven methyl groups at 80.98 (s, H\({}_{3}\)-23), 0.79 (s, H\({}_{3}\)-24), 0.83 (s, H\({}_{3}\)-25), 1.03 (s, H\({}_{3}\)-26), 1.03 (s, H\({}_{3}\)-27), 0.81 (s, H\({}_{3}\)-28), and 1.66 (s, H\({}_{3}\)-30). The predominant characteristic of the lupane class of triterpenoid was found as a pair of singlets at \(\delta\)4.57 and 84.69 with \({}^{1}\)H for each singlet due to the terminal isopropenyl moiety or the vinylic proton located at carbon 29. Meanwhile, the presence of a doublet with one proton intensity found at \(\delta\)3.18 (dd, J = 4.8 Hz, H-3) was caused by the proton attached to the carbon bearing the hydroxyl group or an oxymethine proton located at C-3, which is identical as reported [10]. There was also one proton with a doublet of the triplet located at \(\delta\)2.37, which was assigned to 19\(\beta\)-H orientation, similar to the reported literature data [14]. The \({}^{1}\)H-NMR spectrum for the isolated compound can be found in Supplemental Data S1A. Figure 3: GC-MS profiles of the (**A**) isolated white amorphous compound from the roots of _C. mutans_, (**B**) purchased authentic lupeol, and (**C**) isolated compound spiked with authentic lupeol. Meanwhile, the \({}^{13}\)C-NMR spectrum (150 MHz, CDCl\({}_{3}\)) of the isolated compound exhibited signals at $109.36 (C-29) and $78.99 (C-3), which were the characteristics of protons and carbons of a methylene and oxymethine group at the positions 3 and 29 for lupeol [15]. In addition, another distinguishable characteristic of triterpenoid was found at $150.91 for the presence of carbon C-20 [15]. The \({}^{13}\)C-NMR also revealed carbon intensity at $38.86 (C-1), 27.41 (C-2), 38.74 (C-4), 55.33 (C-5), 18.03 (C-6), 34.31 (C-7), 40.85 (C-8), 50.46(C-9), 37.18 (C-10), 20.95 (C-11), 25.16 (C-12), 38.07 (C-13), 42.84 (C-14), 27.47 (C-15), 35.60 (C-16), 47.99 (C-17), 48.32 (C-18), 47.99(C-19), 29.87 (C-21), 38.86 (C-22), 27.47 (C-23), 15.40 (C-24), 16.13 (C-25), 16.00 (C-26), 14.57 (C-27), 18.34 (C-28), and 19.33 (C-30), which were identical to the reported literature data by [12; 15; 16]. The \({}^{13}\)C-NMR spectrum for the isolated compound can be found in Supplemental Data S1B. The Isolated Lupeol from C. mutans Roots Demonstrates Comparable Anti-proliferative Activity with Authentic Lupeol and Camphtothecin  
The anti-proliferative activity of the lupeol isolated from the C. nutans roots was tested against human breast cancer cell line MCF-7 using MTT assay, and the result was compared with the anti-proliferative activity of authentic standard lupeol (Figure 4).