Journal of Food and Nutrition Research
ISSN (Print): 2333-1119 ISSN (Online): 2333-1240 Website: Editor-in-chief: Prabhat Kumar Mandal
Open Access
Journal Browser
Journal of Food and Nutrition Research. 2015, 3(1), 34-39
DOI: 10.12691/jfnr-3-1-6
Open AccessArticle

Oleanolic acid Separation From Grape Skins by Aqueous Two-phase Extraction and Estimate Its Antioxidant Activity

Xifeng Zhang1, Fenqin Zhang1, Guanghong Luo2, , Shenghui Yang2 and Danxia Wang2

1The College of Agriculture and Biotechnology (CAB), Hexi University, Zhangye, P.R. China

2Kaiyuan Bio-tech Development Center, Hexi University, Zhangye, P.R. China

Pub. Date: January 06, 2015

Cite this paper:
Xifeng Zhang, Fenqin Zhang, Guanghong Luo, Shenghui Yang and Danxia Wang. Oleanolic acid Separation From Grape Skins by Aqueous Two-phase Extraction and Estimate Its Antioxidant Activity. Journal of Food and Nutrition Research. 2015; 3(1):34-39. doi: 10.12691/jfnr-3-1-6


The aim of the current study is to focus on separating the oleanolic acid (OA) from previously prepared aqueous two-phase extraction (ATPE) of grape skins. Several different influential extraction parameters, such as ethanol concentration (v/w), ammonium sulphate concentration (w/w), crude extract concentration (w/w), extraction temperature, and pH, were also investigated. The optimal differential partitioning of OA was achieved in a system (at pH4.0, temperature=25°C)composed of 23% (v/w) ethanol, 18% (w/w) ammonium sulphate, 8% (w/w) crude extract and 41% (w/w) water. The recovery of extracted OA from experiments was determined to be 93.54%. The antioxidant activity of the separated ATPE in relative to VC were, also, investigated in this study at the proper conditions.The ATPE extract showed a relatively high antioxidant ability compared with that of Vitamin C. This proposed extraction technique opens up new possibilities in extraction of other active ingredients in natural plants or biologic samples.

oleanolic acid Aqueous two-phase extraction (ATPE) Ethanol/ammonium sulphate Antioxidant activity

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Torres, J.L.,Varela, B., García, M.T., Carilla, J., Matito,C., Centelles, J. J., and Bobet, R., “Valorization of grape (Vitis vinifera) byproducts. Antioxidant and biological properties of polyphenolic fractions differing in procyanidin composition and flavonol content,” Journal of Agricultural and Food Chemistry, 50 (26). 7548-7555. Nov. 2002.
[2]  Ky, I., Crozier, A., Cros, G., and Teissedre, P. L., “Polyphenols composition of wine and grape sub-products and potential effects on chronic diseases,” Nutrition and Aging, 2 (2). 165-177. Jun. 2014.
[3]  Azuma, A., Yakushiji, H., Koshita, Y., and Kobayashi, S., “Flavonoid biosynthesis-related genes in grape skin are differentially regulated by temperature and light conditions,” Planta, 236 (4).1067-1080. May. 2012.
[4]  Li, Z., Pan, Q., Cui, X., and Duan, C., “Optimization on anthocyanins extraction from wine grape skins using orthogonal test design,” Food Science and Biotechnology, 19 (4).1047-1053. Aug. 2010.
[5]  Pensec, F., Pączkowski, C., Grabarczyk, M., Woźniak, A., Bénard-Gellon, M., Bertsch, C., and Szakiel, A., “Changes in the Triterpenoid Content of Cuticular Waxes during Fruit Ripening of Eight Grape (Vitis vinifera) Cultivars Grown in the Upper Rhine Valley.” Journal of agricultural and food chemistry, 62 (32). 7998-8007. Jul. 2014
[6]  Wang, X., Ye, X. L., Liu, R., Chen, H. L., Bai, H., Liang, X., and Hai, C. X., “Antioxidant activities of oleanolic acid in vitro: possible role of Nrf2 and MAP kinases,” Chemico-biological interactions, 184 (3). 328-337. Mar. 2010.
[7]  Lee, W., Yang, E. J., Ku, S. K., Song, K. S., and Bae, J. S., “Anti-inflammatory effects of oleanolic acid on LPS-induced inflammation in vitro and in vivo,” Inflammation, 36 (1). 94-102. Aug. 2013.
[8]  Ghosh, S., Bishayee, K., and Khuda-Bukhsh, A. R., “Oleanolic acid isolated from ethanolic extract of Phytolacca decandra induces apoptosis in A375 skin melanoma cells: drug-DNA interaction and signaling cascade,” Journal of integrative medicine, 12 (2). 102-114. Mar. 2014.
[9]  Mahapatra, A., Chauhan, N., Patel, D. R., Kalia, N. P., Rajput, V. S., and Khan, I. A., “Synthesis and Antitubercular Activity of Oleanolic Acid Analogs”, Pharmaceutical Chemistry Journal, 48 (1). 39-43. May. 2014.
[10]  Fu, Q., Zhang, L., Cheng, N., Jia, M., and Zhang, Y., “Extraction optimization of oleanolic and ursolic acids from pomegranate (Punica granatum L.) flowers,” Food and Bioproducts Processing. 92 (3). 321-327. Jul. 2014.
[11]  Yang, Y. C., Wei, M. C., Hong, S. J., Huang, T. C., and Lee, S. Z., “Development/optimization of a green procedure with ultrasound-assisted improved supercritical carbon dioxide to produce extracts enriched in oleanolic acid and ursolic acid from Scutellaria barbata D. Don,” Industrial Crops and Products, 49, 542-553. Aug. 2013.
[12]  Pai, S. R., Upadhya, V., Hegde, H. V., Joshi, R. K., and Kholkute, S. D, “New Report of Triterpenoid Betulinic Acid along with Oleanolic Acid from Achyranthes aspera by Reversed-Phase-Ultra Flow Liquid Chromatographic Analysis and Confirmation Using High-Performance Thin-Layer Chromatographic and Fourier Transform-Infrared Spectroscopic Techniques,” JPC-Journal of Planar Chromatography-Modern TLC, 27 (1), 38-41. Feb. 2014
[13]  Ahmad, M. M., and Przybycien, T. M., “Towards optimal aqueous two‐phase extraction system flowsheets for protein purification,” Journal of Chemical Technology and Biotechnology, 88 (1), 62-71. Jan. 2013.
[14]  Wu, X., Liang, L., Zou, Y., Zhao, T., Zhao, J., Li, F., and Yang, L., “Aqueous two-phase extraction, identification and antioxidant activity of anthocyanins from mulberry (Morus atropurpurea Roxb.),”Food Chemistry, 129 (2), 443-453. Nov. 2011.
[15]  Yang, X., Zhang, S., Yu, W., Liu, Z., Lei, L., Li, N., and Yu, Y., Ionic liquid-anionic surfactant based aqueous two-phase extraction for determination of antibiotics in honey by high-performance liquid chromatography. Talanta, 124, 1-6. Jun. 2014.
[16]  Liu, X., Mu, T., Sun, H., Zhang, M., and Chen, J., “Optimisation of aqueous two-phase extraction of anthocyanins from purple sweet potatoes by response surface methodology,” Food chemistry, 141 (3), 3034-3041. Dec. 2013.
[17]  Chen, F., Sun, Y., Zhao, G., Liao, X., Hu, X., Wu, J., and Wang, Z., “Optimization of ultrasound-assisted extraction of anthocyanins in red raspberries and identification of anthocyanins in extract using high-performance liquid chromatography–mass spectrometry,” Ultrasonics Sonochemistry, 14 (6), 767-778. Sep. 2007.
[18]  Wang, B. Z., and XIE H.X., “Determination of concentration of total saponins and oleanolic acid of Rubus sachalinensis at different collecting time,” Chinese Journal of experiment medical formulae, 20 (12), 77-79. Jun. 2014.
[19]  Liu, S. C., Lin, J. T., Wang, C. K., Chen, H. Y., and Yang, D. J., “Antioxidant properties of various solvent extracts from lychee (Litchi chinenesis Sonn.) flowers,” Food Chemistry, 114 (2). 577-581. May. 2009
[20]  Wang, Y., Han, J., Xu, X. H., Hu, S. P., and Yan, Y. S., “Partition behavior and partition mechanism of antibiotics in ethanol/2-propanol-ammonium sulfate aqueous two-phase systems,” Separation and Purifition Technology, 75 (3). 352-357. Nov. 2010
[21]  Yan, J. K., Ma, H. L., Pei, J. J., Wang, Z. B., and Wu, J. Y., “Facile and effective separation of polysaccharides and proteins from Cordyceps sinensis mycelia by ionic liquid aqueous two-phase system,” Separation and Purification technology. 135. 278-284. Oct. 2014.
[22]  Zafarani-Moattar, M. T., & Hamzehzadeh, S., “Phase diagrams for the aqueous two-phase ternary system containing the ionic liquid 1-butyl-3-methylimidazolium bromide and tri-potassium citrate at T=(278.15, 298.15, and 318.15) K,” Journal of Chemical & Engineering Data, 54 (3), 833-841. Dec. 2008.