Elsevier

Journal of Ethnopharmacology

Volume 156, 28 October 2014, Pages 88-96
Journal of Ethnopharmacology

Research Paper
Quantification of polyphenols and evaluation of antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone–water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava

https://doi.org/10.1016/j.jep.2014.07.031Get rights and content

Abstract

Ethnopharmacological relevance

Vast numbers of plant species from northeastern Brazil have not yet been phytochemically or biologically evaluated.

Aim of the study

The goal of this work was to obtain, characterize and show the antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone–water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava.

Materials and methods

The plant material (100 g) was dried, and the crude extracts were obtained by using turbo-extraction (10%; w/v) with water or acetone:water (7:3, v/v) as the extraction solvent. High-performance liquid chromatography (HPLC) methods were used to screen the crude extracts for hydrolysable tannins (gallic acid) and condensed tannins (catechins). The antibacterial activity was evaluated by agar-diffusion and microdilution methods against Gram-positive strains (Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis INCQS 00016, Enterococcus faecalis ATCC 29212 and a clinical isolate of methicillin-resistant Staphylococcus aureus) as well as Gram-negative strains (Escherichia coli ATCC 25922, Salmonella enteritidis INCQS 00258, Shigella flexneri and Klebsiella pneumoniae). To evaluate the anti-inflammatory activity, a leukocyte migration model was used. Analgesic activity was determined by the hot plate test and the acetic acid-induced abdominal writhing test. Data were analyzed by analysis of variance (ANOVA) at a significance level of 5%.

Results

Parapiptadenia rigida presented the highest amount of total polyphenols (35.82±0.20%), while the greatest catechin content was found in the acetone–water extract of Psidium guajava (EAWPg; 1.04 μg/g). The largest amounts of catechins were found in the aqueous extract of Libidibia ferrea (EALf; 1.07 μg/g) and the acetone–water extract of Parapiptadenia rigida (EAWPr; 1.0 μg/g). All extracts showed activity against Gram-positive bacteria. The aqueous and acetone–water extracts of Psidium guajava showed the greatest inhibition zones in the agar diffusion tests. In the evaluation of the minimum inhibitory concentration (MIC), the most susceptible Gram-positive bacterium was Staphylococcus epidermidis and the most susceptible Gram-negative bacterium was Shigella flexneri. EAPg and EAWPg showed the greatest MIC values. All extracts were significant inhibitors of leukocyte migration (p<0.05). Using the writhing test, significant analgesic activity was found for EAPr (50 mg/kg), EAWPr (100 mg/kg and 200 mg/kg) and EAWPg (50 mg/kg) (p<0.05).

Conclusions

Thus, the appropriate extraction procedure preserves the chemical components such as gallic acid and catechin, and showed antimicrobial, anti-inflammatory and analgesic properties.

Introduction

Brazil is one of the countries with the greatest diversity of plants with different biomes, and these plants may have therapeutic potential. The semi-arid Caatinga biome that exists in northeastern Brazil has been little studied (de Albuquerque et al., 2007). Therefore, it is vital to study the utilitarian plant species from semi-arid regions of northeastern Brazil because new economic alternatives can ensure the livelihood and maintenance of these species. Among the species popularly used in the northern and semi-arid northeastern regions of Brazil are Libidibia ferrea, Parapiptadenia rigida and members of the genus Psidium (Sampaio et al., 2009, Costa et al., 2012, De Souza et al., 2012).

Libidibia ferrea (Mart.) L. P. Queiroz belongs to the Caesalpinaceae family and, as a medicinal plant, its use has been reported in the treatment of virus, inflammation and diarrhea (Pereira et al., 2012, de Medeiros et al., 2013, Lopes et al., 2013). Its popular name is ironwood, and it is composed of the following phenolic compounds: condensed tannins, catechins and hydrolysable tannins such as gallic acid and ellagic acid (Frasson et al., 2003, Vasconcelos et al., 2011).

Parapiptadenia rigida (Benth.) Brenan belongs to the Fabaceae family and is popularly known as red angico. As a medicinal plant, the bark is used in the treatment of inflammation, gastrointestinal disorders, tumors and chronic diseases (Souza et al., 2004). The bark has phenolic compounds such as tannins, catechin derivatives and proanthocyanidins (Schmidt et al., 2011).

Psidium guajava L. belongs to the Myrtaceae family and is popularly known as guava. It is present in all tropical and subtropical regions of the world, including Latin America, Europe, Asia and Africa (Gutierrez et al., 2008). In northeastern Brazil, its leaves are used to treat dysentery, digestive problems, inflammation, gingivitis and throat afflictions (de Albuquerque et al., 2007). Its bark and leaves have tannins, flavonoids, saponins and alkaloids (Tona et al., 2000). The biological characteristics of Psidium guajava include antidiarrheal, amoebicidal, spasmolytic, antibacterial and antifungal activities (Birdi et al., 2010, Rahim et al., 2010).

In this context, most of the plant compounds that have been found to be medicinally useful and interesting tend to be secondary metabolites. The choice of extraction solvent strongly affects the yield and the number of metabolites obtained. Different solvent systems have been used to extract secondary metabolites from plant materials because their extraction efficacy depends on their chemical nature. For example, ethyl alcohol extracts a large number of flavonoids; while aqueous mixtures of ethanol or acetone have been used to extract polyphenols from plant material (Garcia-Salas et al., 2010).

Thus, the aim of this work was to obtain and quantitate the polyphenol content and verify the antimicrobial, analgesic and anti-inflammatory activities of aqueous and acetone–water extracts of Libidibia ferrea, Parapiptadenia rigida and Psidium guajava.

Section snippets

Plants

Samples of Libidibia ferrea bark, Parapiptadenia rigida bark and Psidium guajava leaves were collected in the states of Pernambuco, Paraíba and Rio Grande do Norte, respectively. Voucher specimens were deposited at the Agronomy Institute of Pernambuco (IPA), the Department of Botany at the Federal University of Pernambuco (UFPE) and the Department of Botany at the Federal University of Rio Grande do Norte (UFRN), under numbers 88145, 83115 and 8214, respectively. The plant names have been

Phytochemical analysis

TLC for chemical identification of the constituents of the crude extracts from Libidibia ferrea bark, Parapiptadenia rigida bark and Psidium guajava leaves revealed the presence of condensed tannins (catechins) and hydrolysable tannins (gallic acid).

Estimation of TPC

The TPC of the crude extracts were calculated. Comparison of the average crude extract yields showed that the yields from the aqueous extracts were all less than the yields from the acetone–water extracts (Table 1).

HPLC analysis

The chromatographic fingerprints

Discussion

In our study, we found that the method of extraction, with water or an acetone–water mixture, maintained the presence of polyphenols, including total polyphenols, gallic acid and catechin, confirming previous reports in which these components have been identified in the plant species used in this study (Gasparin Verza et al., 2007, Schmidt et al., 2010, Vasconcelos et al., 2011, Chang et al., 2013).

The literature regarding Libidibia ferrea and Parapiptadenia rigida is scarce, and this study may

Conclusion

In conclusion, the choice of extraction solvent is important to preserve the chemical components, such as gallic acid and catechins, which may be related to the antimicrobial, anti-inflammatory and analgesic activities found in these extracts.

Acknowledgments

This work was supported by grants from CNPQ (483870/2011-0; 480128/2012-0; 302113/2012-6) and FACEPE (IBPG-0423-4.03/11, APQ-1296-4.03-12).

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