Contents lists available at ScienceDirect Biochemical Systematics and Ecology journal homepage: www.elsevier.com/locate/biochemsyseco Flavonoids and other compounds from Dioclea virgata (Rich.) Amsh Clayton Queiroz Alves a , Jorge Mauricio David b,* , Juceni Pereira David c , Anake Kijjoa d a Departamento de Ciências Exatas, Universidade Estadual de Feira de Santana, Feira de Santana, Bahia, Brazil b Instituto de Química, Universidade Federal da Bahia, Salvador, Bahia, Brazil c Faculdade de Farmácia, Universidade Federal da Bahia, Salvador, Bahia, Brazil d ICBAS - Instituto de Ciências Biomédicas de Abel Salazar, Rua de Jorge Viterbo Ferreira, 228, Porto 4050-313, Portugal ARTICLE INFO Keywords: Dioclea virgata Flavonoids Megastigmane Benzoic acid derivatives 1. Subject and source Dioclea virgata (Rich.) Amsh (synonym: D. lasiocarpa Benth.) is a climbing plant popularly known in Brazil as “cipó de anauerá”, “cipó pixuma”, “feijão bravo”, “feijãorana”, “mucuna” and “mucunã” (Silva, 2004). This plant is present in South and Central America, and it is used in folk medicine to treat fever and malaria (Agra et al., 2007). Plant material was collected at the campus of Universidade Federal da Bahia (S 13°00.018′ W 38°30.411′), Salvador (BA), Brazil, an area of re- maining Atlantic Forest. The plant was identified by Prof. Dr. Luciano P. de Queiróz, and a voucher specimen (HUEFS # 115979) was deposited in the Herbarium of Biology Department of Universidade Estadual de Feira de Santana (UEFS), Bahia, Brazil. 2. Previous work The isolation and structural study of a lectin from the seeds of D. virgata was previously described (Nóbrega et al., 2012). No other pre- vious reports concerning secondary metabolites from D. virgata are available. However, some bioactive compounds have been isolated from other species of this genus. For example, the antimicrobial dicli- midazole (Oladosu et al., 2010) and flavonoids (Oladimeji et al., 2018) were isolated from D. reflexa Hook. f. and D. grandiflora Mart. ex Benth. (Jenkins et al., 1999) and antioxidant terpenoids and proanthocyani- dins from D. lasiophylla Mart. ex Benth. (Barreiros et al., 2000; David et al., 2004). Furthermore, pharmacological study with the chloroform extract of D. virgata leaves exhibited in vitro antinociceptive activity (Mota et al., 2011). 3. Present study The leaves (1.7 kg) and stems (2.0 kg) of D. virgata were dried, powdered and exhaustively extracted with MeOH, separately, at room temperature. The methanolic solutions were evaporated under reduced pressure to generate crude methanol extracts, which were dissolved in a mixture of MeOH:H 2 O (8:2 v/v, 800 ml) and submitted to a partition with chloroform (5 × 300 ml). After complete evaporation, the CHCl 3 soluble fraction was dissolved in MeOH (500 ml) and extracted with hexane (5 × 300 ml). The hydromethanolic phase was evaporated under reduced pressure, dissolved in water (500 ml) and partitioned with EtOAc (5 × 300 ml). The fractions were evaporated to dryness, yielding 11.3 g, 8.7 g and 12.9 g of hexane, chloroform and EtOAc so- luble fractions of stems, respectively, and 32.9 g, 23.0 g and 13.6 g of hexane, chloroform and EtOAc soluble fractions of leaves, respectively. The crude hexane extract (11.3 g) of the stems was applied on a column of silica gel (55 g), and eluted with hexane:EtOAc (95:05 > 0:100), wherein 14 fractions of 100 ml each were collected and then pooled based on their TLC profiles into 10 fractions. Fractions 3, 4 and 6 were identified, based on 1 H and 13 C NMR spectral analysis and comparison of their spectroscopic data with those reported in the literature, as lupenone (1, 360 mg) (Luz et al., 2010), lupeol (2, 50 mg) (Souza et al., 2001) and a mixture of β-sitosterol and stigmasterol (3 and 4, 30 mg) (Greca et al., 1990), respectively. The chloroform extract (8.7 g) of stems was also submitted to silica gel CC (45 g) and eluted with mixtures of CHCl 3 :MeOH (95:5 > 0:100), to yield 17 fractions (100 ml each), which were combined into 8 fractions based on TLC profile. Fr 3 (250 mg) was applied on a Sephadex LH-20 column (15 g) and eluted with https://doi.org/10.1016/j.bse.2018.03.011 Received 12 December 2017; Received in revised form 27 March 2018; Accepted 28 March 2018 * Corresponding author. E-mail address: jmdavid@ufba.br (J.M. David). Biochemical Systematics and Ecology 78 (2018) 43–45 0305-1978/ © 2018 Published by Elsevier Ltd. T