*Corresponding author Email address: gorawit.yu@wu.ac.th Songklanakarin J. Sci. Technol. 42 (5), 1084-1090, Sep. - Oct. 2020 Original Article A green and effective method using oils to remove chlorophyll from Chromolaena odorata (L.) R.M. King & H. Rob Suppalak Phaisan 1 , Gorawit Yusakul 1, 2* , Attapon Sakdamas 1 , Nattapon Taluengjit 1 , Seiichi Sakamoto 3 , and Waraporn Putalun 4, 5 1 School of Pharmacy, Walailak University, Tha Sala, Nakhon Si Thammarat, 80160 Thailand 2 Drug and Cosmetics Excellence Center, Walailak University, Tha Sala, Nakhon Si Thammarat, 80160 Thailand 3 Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, 8128582 Japan 4 Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand 2 Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology, Khon Kaen University, Mueang, Khon Kaen, 40002 Thailand Received: 8 February 2019; Revised: 7 July 2019; Accepted: 17 July 2019 Abstract Chromolaena odorata (L.) R.M. King & H. Rob has excellent wound healing effects. Therefore, it is useful for cosmeceutical purposes; however, the dark-greenish appearance of its extract limits its attractiveness for product development. This study aimed to develop an oil-based system for chlorophyll removal. Dried ethanolic extracts were dissolved in different aqueous EtOH solvents, and then the chlorophyll was partitioned to the oil phase. The efficiency of chlorophyll removal was monitored using a spectrophotometer. The recoveries of flavonoid and phenolic contents and the antioxidant activity were determined after the treatment. When the solutions of extract were prepared in 25% (v/v) - 75% (v/v) EtOH in water, the efficacy of chlorophyll removal by oil was higher than 85%. A higher concentration of EtOH resulted in a lower chlorophyll removal efficiency; however, the recoveries of phenolic and flavonoid contents and antioxidant activity improved. Palm oil showed efficiency higher than hexane for chlorophyll removal and high recoveries of the beneficial phytochemicals. This system can be applied in the fields of natural products-based health product development and in phytochemical studies. Keywords: Chromolaena odorata, chlorophyll removal, oil, antioxidant, flavonoids 1. Introduction Chromolaena odorata (L.) R.M. King & H. Rob, called Siam weed (“Sap suea” in Thai), has received much attention due to its various pharmaceutical activities. In Thailand and other tropical countries, fresh leaves of C. odorata have traditionally been used to stop bleeding and enhance wound healing. Scientific investigations revealed that the extract of C. odorata exhibited various activities involved in wound healing. For the reasons mentioned above, the extract of C. odorata may be useful as an active ingredient of wound healing and cosmeceutical products. The leaves of C. odorata contain various classes of compounds such as flavonoids (Phan et al., 2001), phenolics (Phan et al., 2001), alkaloids (Biller, Boppré, Witte, & Hart mann, 1994), and essential oils (Pisutthanan et al., 2006). An