1. 150 mL of DPPH(2,2-diphenyl- 1-picrylhydrazyl) 250 mM solution. Incubation for 30 mins Remaining percentage of DPPH was measured at 490 nm on an ELISA reader. 50 mL of each dilution was transfered into a 96- well microplate Comparison of Various Extraction Methods for the Evaluation of Bioactivities of Vitex negundo collected from the lowland subtropical habitat in southern Taiwan Chuan-Jhao Jhuang 1 ， Yu-Wen Liao 2 ， Po-Chuen Shieh 1* 1 Department of Pharmacy, Tajen University,Yanpu,Pingtung 907,Taiwan 2 Department of vogue and beauty, Tajen University,Yanpu,Pingtung 907,Taiwan The effect of extraction solvents and temperatures on extraction yields of various functional compounds from plant materials was investigated. Taiwan’s lowland subtropical harbors rare species of plants.Some of these plants are used in the traditional medicine to treat a widerange of conditions including bacterial, fungal and viral diseases. We have evaluated Vitex negundo (Verbenaceae) that collected from the lowland subtropical rainforest habitat in southern Taiwan to determine total polyphenol content and antioxidant activity (by 2,2-diphenyl-1,1-picrylhydrazyl DPPH free radical scavenging assay against -tocopherol). The contents of polyphenols in dry plant materials ranged from 2.3 to 155 mg/g in which are able to eliminate significantly. DPPH in 0.5 h indicating that the analyzed plant possessfree radical scavenging activity. Our research indicates that Vitex negundo has great potential in scavenging free radicals and the plant can be a vital source of antioxidant phytochemicals. Diluted volatile oil of Vitex negundo by multiple as 1000ppm, 500ppm, 250ppm and 125ppm, 62.5ppm respectively in 0.9%NaCl. added 300 μL H 2 O 2 and 100 μL Vitex negundo diluted solution into luminescence analyzer to make the first detection 1ml luminal was adding at 60 seconds and made measurement until 300 seconds. Scientific name: Vitex negundo L. Family name: Verbenaceae Verbenaceae Generic name: Vitex (Vitex) Distribution: tropical Asia, Polynesia, tropical East Africa and Taiwan. Accordance with the above steps repeated 3 times for each sample.(n=3) Fig.1 DPPH experimental procedures Fig.3 DPPH r esidual rate Fig.2 DPPH calibration curve Fig.4 chemiluminescence experimental procedures Fig.5 Vitamin C and Vitex negundo free radical inhibition rate of H 2 O 2 Fig.6 Vitex negundo’s chemiluminescence analysis chart CL intensity counts Conclusion This study was based on using different extraction methods and current conditions, extraction of Vitex negundo was determined by experiment of antioxidant, include DPPH analysis and Luminol chemiluminescence test. We found that Vitex negundo extracts had free radical scavenging capacity(Fig3). It showed that the free radical scavenging ability 1000ppm of scavenging extract of free radicals had close ability with 10ppm of vitamin C,therefore,we assumed that Vitex negundo was good in antioxidant. In the future, the research on purified of Vitex negundo extracts, will able to make a series of antioxidant and scavenging free radicals and further to study in free radical scavenging capacity of rat organs analysis.