FULL COMMUNICATION Identification of Antioxidant and Antimicrobial Compounds of Lippia Species by Bioautography Rodrigo Luiz Fabri, Mauro Silva Nogueira, Jussara dos Reis Moreira, Maria Lu ´cia Morcef Bouzada, and Elita Scio Department of Biochemistry, Institute of Biological Sciences, Universidade Federal de Juiz de Fora, Campus Universita´rio, Martelos, Juiz de Fora, Minas Gerais, Brazil. ABSTRACT The methanolic extracts of the leaves of Lippia species (L. pseudo-thea, L. hermannioides, L. alba, L. rubella, and L. sidoides) were tested for their antibacterial, antifungal, and antioxidant activity. Cytotoxicity was determined by using brine shrimp lethality bioassay. Phytochemical screening was also performed. The extracts showed a minimum inhibitory concentration (MIC) ranging from 78 to 5000 mg=mL for antibacterial activity against at least 2 species of bacteria, although none was active against Escherichia coli. Antifungal activity was found only for L. pseudo-thea (MIC, 625 mg=mL for Candida albicans) and L. sidoides (MIC, 625 mg=mL for both C. albicans and C. neoformans). The bioautography showed that flavonoids and coumarins are responsible for the antioxidant activity of the extracts and that the antimicrobial properties are due to flavonoids and terpenoids. The cytotoxic activity was stronger for L rubella extract. To our knowledge, this is the first report of the biological and chemical constituents of L. pseudo-thea, L. hermannioides, and L. rubella. KEY WORDS: antimicrobial  antioxidant  bioautography  Lippia  polyphenols INTRODUCTION T he genus Lippia (Verbenaceae) is widely distributed in tropical and subtropical regions of the Americas and Africa, and it consists of approximately 200 species of herbs, shrubs, and small trees. 1 In Brazil, the genus Lippia is re- presented by nearly 120 species conspicuous for their ap- pearance during their short blooming period and their generally strong and pleasant fragrance. 2 The leaves from most of these species are used as seasoning for food prepa- ration. 3 The most common use of Lippia species is for treating respiratory disorders. In Central and South America (Guatemala, Venezuela, and Brazil), they are also used as a remedy for colds, influenza, bronchitis, coughs, and asth- ma. 3–6 Biological activity and economic importance of some Lippia species, as well as their chemical constituents, have been reported in previous studies. 7 The aim of this study was to evaluate the antioxidant and antimicrobial activities of the methanolic extracts of the leaves obtained from 5 Lippia species popularly used in traditional medicine. Cytotoxicity was determined by using brine shrimp lethality bioassay, and phytochemical screen- ing was performed. The total amount of phenolics and fla- vonoids was determined. To our knowledge, this is the first report of the biological and chemical constituents of L. pseudo-thea, L. hermannioides, and L. rubella. MATERIALS AND METHODS Plants The Lippia species were collected in Juiz de Fora, Minas Gerais, Brazil, in July 2005. Voucher specimens were deposited at the Herbarium Leopoldo Krieger (CESJ) of Federal University of Juiz de Fora. The species used in this study were L. pseudo-thea Schauer [45171], L. hermannioides Cham. [46088], L. alba Mill. (N.E.) [46177], L. rubella Moldenke [46178], and L. sidoides Cham. [46180]. Preparation of the extract The leaves (50 g each) were powdered and macerated with methanol (3200 mL) for 5 days at room temperature. After evaporation of the solvent under reduced pressure at 458C, the respective methanolic extracts were obtained. All the extracts were kept in tightly stoppered bottles under refrigeration until used for the biological testing and phy- tochemical screening. Phytochemical screening A portion of each extract that was subjected to the bio- logical screening was used to identify the major secondary metabolites according to protocols described elsewhere. 8 Manuscript received 28 May 2010. Revision accepted 30 September 2010. Address correspondence to: Elita Scio,Departamento de Bioquı´mica–ICB–Universidade Federal de Juiz de Fora, Campus Universita´rio, Martelos, Juiz de Fora, MG, Brazil, E-mail: elita.scio@ufjf.edu.br JOURNAL OF MEDICINAL FOOD J Med Food 14(0) 2011, 1–7 #Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition DOI: 10.1089=jmf.2010.0141 1