Natural Product Research Formerly Natural Product Letters

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Characterization and identification of chemical constituents of Ocotea paranaensis Brotto, Baitello, Cervi, & E.P. Santos and their biological properties Caroline Gribner , Katlin Suellen Rech , Paula Francislaine Moura , Ana Angélica Ruscheweyh Rigoni , Larissa Junqueira Gatto , Andressa Veiga , Vitor Alberto Kerber , Marilis Dallarmi Miguel , Obdulio Gomes Miguel & Josiane de Fátima Gaspari Dias To cite this article: Caroline Gribner , Katlin Suellen Rech , Paula Francislaine Moura , Ana Angélica Ruscheweyh Rigoni , Larissa Junqueira Gatto , Andressa Veiga , Vitor Alberto Kerber , Marilis Dallarmi Miguel , Obdulio Gomes Miguel & Josiane de Fátima Gaspari Dias (2020): Characterization and identification of chemical constituents of Ocotea�paranaensis Brotto, Baitello, Cervi, & E.P. Santos and their biological properties, Natural Product Research, DOI: 10.1080/14786419.2020.1826476 To link to this article: https://doi.org/10.1080/14786419.2020.1826476

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NATURAL PRODUCT RESEARCH https://doi.org/10.1080/14786419.2020.1826476

SHORT COMMUNICATION

Characterization and identification of chemical constituents of Ocotea paranaensis Brotto, Baitello, Cervi, & E.P. Santos and their biological properties Caroline Gribner , Katlin Suellen Rech , Paula Francislaine Moura Ana Ang
elica Ruscheweyh Rigoni , Larissa Junqueira Gatto , Andressa Veiga , Vitor Alberto Kerber, Marilis Dallarmi Miguel , Obdulio Gomes Miguel and Josiane de F
atima Gaspari Dias

,

Pharmaceutical Sciences Graduate Program, Federal University of Paran
a, Curitiba, PR, Brazil ABSTRACT

ARTICLE HISTORY

The growing interest in the discovery of new compounds from medicinal plants justifies the study of phytochemical components of these plants and their biological activities. The genus Ocotea exhibits a variety of pharmacological, antimicrobial and antioxidant effects. The aim of the present study was to evaluate the antioxidant potential and antimicrobial properties of the ethyl acetate fraction of Ocotea paranaensis leaves. Isolation and identification of the phenolic compounds from the fraction was also carried out. The isolated compounds were characterized by one and two-dimensional nuclear magnetic resonance analysis and identified as (
) epicatechin (1), quercetin (2), kaempferol (3) and hyperin (4). The ethyl acetate fraction of Ocotea paranaensis leaves demonstrated considerable antioxidant potential. The observed minimum inhibitory concentration of 500 mg/mL was classified as a moderate antibacterial activity against Staphylococcus aureus. Findings from this study demonstrate the utility of this plant as a potential source of antioxidant and antimicrobial agents.

Received 27 April 2020 Accepted 5 September 2020 KEYWORDS

Lauraceae; Ocotea paranaensis; phytochemical characterization; antimicrobial activity; biological properties; leaves

CONTACT Caroline Gribner [email protected] Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2020.1826476. ß 2020 Informa UK Limited, trading as Taylor & Francis Group

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1. Introduction Ocotea paranaensis Brotto, Baitello, Cervi, and E.P. Santos taxonomically belongs to the Lauraceae family and is widely distributed in tropical and subtropical areas of the Americas, Africa, Canary Islands, Madagascar and Eastern Asia. This family is composed of approximately 55 genera and 3000 species. The genus Ocotea is among the largest genus of Lauraceae, and contains approximately 350 species (Baitello 2001; Werff 2013). Ocotea species exhibits a variety of pharmacological effects, including anti-inflammatory (Ocotea bullata and Ocotea macrophylla) (Zschocke et al. 2000; Coy-Barrera and Cuca-Su
arez 2011), antinociceptive (Ocotea puberula) (Montrucchio et al. 2012), antioxidant (Ocotea bullata) (Ogundajo et al. 2018) and antimicrobial properties (Ocotea odorifera) (Gontijo et al. 2017). Several chemical constituents have been discovered in the genus Ocotea including alkaloids, flavonoids, sesquiterpenes and saponins (Salleh and Ahmad 2017). Ocotea paranaensis Brotto, Baitello, Cervi, and E.P. Santos was discovered in the Atlantic Forest in Paran
a State, a place from where it derived its name (Brotto et al. 2010). In the present study, we isolate four flavonoids and evaluate the antioxidant potential and antimicrobial properties of the ethyl acetate fraction of the leaves of Ocotea paranaensis Brotto, Baitello, Cervi, and E.P. Santos.

2. Results and discussion 2.1. Structural elucidation Thin-layer chromatography (TLC) revealed the presence of phenolic compounds in the ethyl acetate fraction of the leaves and in the precipitate obtained during partitioning.

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Figure 1. Structures of compounds 1–4.

This justifies the characterization and identification of the chemical constituents of plant in this study. Four compounds were isolated and identified from the leaves of Ocotea paranaensis Brotto, Baitello, Cervi, and E.P. Santos for the first time (Figure 1). These phenolic compounds were identified as (–) epicatechin (1), quercetin (2), kaempferol (3), and hyperin (3-O-b-D-galactopyranosil quercetin) (4). Compound 1, obtained as yellow crystals, was subjected to single-crystal X-ray diffraction analysis. The parameters obtained for the unit cell were compared with the data from the Cambridge Crystallographic Data Centre, deposit number 1130393 (Table S1, Supplementary Material). The structures of compounds 2, 3, and 4 were determined using one- and two-dimensional nuclear magnetic resonance (Table S2, Supplementary Material), and the data were compared with the literature (Santos et al. 2005; Salas-Moreno 2014; Lin et al. 2016). The identified bioactive compounds are consistent with previous studies on the genus Ocotea. Epicatechin was identified in the leaves of Ocotea notata (Garrett et al. 2012). Quercetin was identified in the leaves of Ocotea minarum (Rodrigues et al. 2019) and

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Table 1. Antioxidant potential of Ocotea paranaensis Brotto, Baitello, Cervi & E.P. Santos leaves. Mean ± SD Sample DPPH IC50 (mg/mL) ABTSþ (mmol TE/mg) FRAP (mgM ferrous sulfate/g)

FEAL

Ascorbic acid

BHT

Rutin

2.38 ± 0.05a 3.44 ± 0.27c 8.52 ± 0.11b

5.80 ± 0.04b 4.42 ± 0.22d 17.76 ± 0.13d

13.93 ± 0.64d 1.2 ± 0.06a 3.70 ± 0.14a

8.88 ± 0.07c 2.5 ± 0.12b 9.09 ± 0.10c

IC50: inhibitory concentration 50%, SD (standard deviation), TE (Trolox equivalent); means followed by equal letters do not differ according to the Tukey test (p < 0.05).

Ocotea notata (Garrett et al. 2012), and kaempferol was identified in the leaves of Ocotea notata (Garrett et al. 2012). Hyperin was identified in the leaves of Ocotea corymbosa (Batista et al. 2010).

2.2. Total phenolic content The gallic acid equivalent (GAE) of the ethyl acetate fraction of the leaves (FEAL) of Ocotea paranaensis Brotto, Baitello, Cervi & E.P. Santos was quantified as 615.6 mg per gram of sample (GAE/g). According to Chew et al. (2011), this value indicates a high total phenol content in the analysed fraction, which is consistent with the isolation of flavonoids. The total amount of phenolics for FEAL was higher than that for the other species of the same genus. Silva et al. (2017) reported a gallic acid equivalent of 360.04 mg GAE/g in Ocotea lancifolia leaves, while Pereira et al. (2019) reported 72.20 mg GAE/g in Ocotea notata leaves, and Ogundajo et al. (2018) reported 8.97 mg GAE/g in the stem bark of Ocotea bullata.

2.3. Antioxidant properties For the reduction of 2,2-diphenyl-1-picrylhydrazyl (DPPH), the IC50 value for the extract of FEAL was 2.38 mg/mL, indicative of greater antioxidant potential when compared with the IC50 values of the analysed standards (ascorbic acid, butylated hydroxytoluene (BHT), and rutin) (Table 1). The IC50 of the ethyl acetate fraction of the leaves of Ocotea lancifolia was 730 mg/mL (Silva et al. 2017). Ocotea bullata stem bark presented an IC50 of 1390 mg/mL (Ogundajo et al. 2018). The Trolox equivalent (TE) determined from the ABTSþ assay was 3.44 mmol TE/mg for FEAL, which is higher than that of BHT (1.2 mmol TE/mg) and rutin (2.5 mmol TE/mg). Studies of the same genus showed similar results; for Ocotea bullata, the IC50 value for the ethyl acetate extract of the mature stem bark was lower than that of the gallic acid control (IC50 of 270 mg/ mL and 3240 mg/mL, respectively) (Ogundajo et al. 2018). In the determination of Fe3þ-Fe2þ reduction in the presence of the FRAP reagent, the reducing ability of FEAL (8.52 mgM ferrous sulfate/g) was greater than that of BHT (3.70 mgM ferrous sulfate/g) (Table 1). The IC50 value for the ethyl acetate extract of the mature stem bark of Ocotea bullata was similar to that of the gallic acid control (IC50 of 2360 mg/mL and 3210 mg/mL, respectively) (Ogundajo et al. 2018).

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Table 2. Minimum inhibitory concentration (MIC) of fraction ethyl acetate of leaves of Ocotea paranaensis Brotto, Baitello, Cervi & E.P. Santos. Microorganism Candida albicans Pseudomonas aeruginosa Staphylococcus aureus Escherichia coli

MIC (lg/mL) >1000 >1000 500 >1000

2.4. Antimicrobial activity Ethyl acetate fraction of the leaves exhibited bacteriostatic action. The minimum inhibitory concentration (MIC) results were indicative of moderate antibacterial activity (MIC ¼ 500 mg/mL) for Staphylococcus aureus. No bactericidal effect was observed in the test sample. The sample was inactive (MIC > 1000 mg/mL) for Pseudomonas aeruginosa, Escherichia coli and Candida albicans (Table 2). Similarly, the other species of the genus Ocotea exhibited antimicrobial potential. The ethyl acetate extract of the branches and leaves of Ocotea minarum showed moderate activity against Staphylococcus aureus, weak activity against Pseudomonas aeruginosa, and the branch extract was inactive for Candida albicans, while the leaf fraction was inactive for Pseudomonas aeruginosa (Tanaka et al. 2005; Rodrigues et al. 2019). The aqueous extract of Ocotea odorifera leaves showed antibacterial activity against the strain of Staphylococcus aureus, which was correlated with the content of flavonoids isolated from this species (Gontijo et al. 2017). The moderate antibacterial activity of FEAL (MIC ¼ 500 mg/mL) for Staphylococcus aureus €zgec¸ et al. 2005; Taiwo et al. 2019) as may be related to one of the isolated substances (Su well as the synergism of these compounds with other compounds. Ethyl acetate fraction of the leaves was shown to be inactive against Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli), which may be related to the structural differences of Gram-negative (the presence of lipopolysaccharides and porins in the membrane) versus Gram-positive bacteria (S. aureus) (Ayres et al. 2008), where the compounds present in Gram-negative bacteria interfere with the action of the bioactive components found in FEAL. The inactivity may also be related to the low concentration of the compounds (in the fraction).

3. Conclusion All isolated compounds (epicatechin, quercetin, kaempferol and hyperin) were detected for the first time in Ocotea paranaensis Brotto, Baitello, Cervi, and E.P. Santos. The ethyl acetate fraction of Ocotea paranaensis Brotto, Baitello, Cervi, and E.P. Santos contains phenolic compounds, which may contribute to the antioxidant potential demonstrated in this study. Our findings contribute to the knowledge base for the phytochemical composition of this species and demonstrate the utility of the plant as a potential source of antioxidant and antimicrobial agents.

Disclosure statement No potential conflict of interest was reported by the author(s).

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Funding This work was carried out with the support of the Coordination of Improvement of Higher Education Personnel – Brazil (CAPES) – Financing Code 001.

ORCID Caroline Gribner http://orcid.org/0000-0002-0179-1284 Katlin Suellen Rech http://orcid.org/0000-0001-6699-2450 Paula Francislaine Moura http://orcid.org/0000-0003-1617-3144 Ana Ang
elica Ruscheweyh Rigoni http://orcid.org/0000-0001-9986-4228 Larissa Junqueira Gatto http://orcid.org/0000-0001-6451-7445 Andressa Veiga http://orcid.org/0000-0001-5470-7317 Marilis Dallarmi Miguel http://orcid.org/0000-0002-1126-9211 Obdulio Gomes Miguel http://orcid.org/0000-0002-2231-9130 Josiane de F
atima Gaspari Dias http://orcid.org/0000-0002-8548-8505

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