Solvent Fractionation and Acetone Precipitation for Crude Saponins from Eurycoma longifolia Extract

Abstract

Eurycoma longifolia is a popular folk medicine in South East Asia. This study was focused on saccharide-containing compounds including saponins, mainly because of their medical potentials. Different organic solvents such as ethyl acetate, butanol, and chloroform were used to fractionate the phytochemical groups, which were consequently precipitated in cold acetone. Solvent fractionation was found to increase the total saponin content based on colorimetric assay using vanillin and sulfuric acid. Ethyl acetate fraction and its precipitate were showed to have the highest crude saponins after acetone precipitation. The samples were shown to have anti-proliferative activity comparable with tamoxifen (IC50 = 110.6 µg/mL) against human breast cancer cells. The anti-proliferative activities of the samples were significantly improved from crude extract (IC50 = 616.3 µg/mL) to ethyl acetate fraction (IC50 = 185.4 µg/mL) and its precipitate (IC50 = 153.4 µg/mL). LC-DAD-MS/MS analysis revealed that the saccharide-containing compounds such as m/z 497, 610, 723, 836, and 949 were abundant in the samples, and they could be ionized in negative ion mode. The compounds consisted of 226 amu monomers with UV-absorbing property at 254 nm, and were tentatively identified as formylated hexoses. To conclude, solvent fractionation and acetone precipitation could produce saccharide-containing compounds including saponins with higher anti-proliferative activity than crude extract against MCF-7 cells. This is the first study to use non-toxic solvents for fractionation of bioactive compounds from highly complex plant extract of E. longifolia.

Anti-Melanoma Activities and Phytochemical Compositions of Sorbus commixta Fruit Extracts

“…It is well known that ethanol (relative polarity 0.654) can dissolve polar compounds, such as sugar, amino acid, glycoside compounds, phenolic compounds with medium polarity, aglycon flavonoids, anthocyanins, terpenoids, flavones, and polyphenols [ 28 ], indicating that EtOH extract is a complex mixture of organic compounds. Based on the liquid–liquid extraction technique, highly polar substances, such as organic acids, polysaccharides, free sugars, and proteins stay in the aqueous phase, whereas other relatively less polar compounds, such as terpenoids and polyphenols, are more soluble in organic solvents [ 29 , 30 ]. To enrich the active compounds in S .…”

Evaluation of Metabolite Profiles of Ginseng Berry Pomace Obtained after Different Pressure Treatments and Their Correlation with the Antioxidant Activity

“…This is an important step in bio-guided assays of new phytochemical ingredients [ 32 ]. Chua et al [ 33 ] reported that highly polar substances, such as organic acids, polysaccharides, and proteins, might remain in the aqueous phase, while the other less polar compounds, including terpenoids, could be segregated into the organic phase. More specifically, the ethyl acetate fraction had the highest levels of saponins, which include ginsenosides.…”

A review of the phytochemical compounds and pharmacological activities of Eurycoma longifolia

“…Besides that, polyphenols, high molecular weight polysaccharides and glycoprotein were also founded in root of E. longifolia (Tsai et al, 2020). Polar to semipolar saponins have been founded in E. longifolia roots (Chua et al, 2019).…”