Phytochemical composition and antimicrobial and toxicological activity of Spondias mombin L. (jobo)

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Phytochemical composition and antimicrobial and toxicological activity of Spondias mombin L. (jobo)

Composición fitoquímica, actividad antimicrobiana y toxicología de Spondias mombin L. (jobo)

 

Elaine Laíse Cavalcanti Clementino1
Delcio de Castro Felismino2
Humberto Silva2
Jocimar Silva Santos1
Ana Cláudia Dantas de Medeiros1
Thiago Pereira Chaves3*

 

1 Universidade Estadual da Paraíba, Campina Grande, PB, Brasil.
2 Departamento de Biologia, Universidade Estadual da Paraíba, Campina Grande, PB, Brasil.
3 Departamento de Ciências Naturais da Universidade Federal do Piauí, Campus Professora Cinobelina Elvas, Bom Jesus, PI, Brasil.

*Autor para la correspondência. Correo electrónico: thiago_pereira@ufpi.edu.br

 

 

ABSTRACT

Introduction: Spondias mombin L. (Anacardiaceae) is used in folk medicine in the semi-arid region of Brazil against a variety of diseases.
Objective: Evaluate the phytochemical profile and biological activity of S. mombin ethanolic extract.
Methods: Ethanolic extract was obtained from S. mombin bark by turbo-extraction and subjected to phytochemical analysis by UV-Vis spectrophotometry. Microbial susceptibility was evaluated by microdilution and toxicity was determined in Artemia salina nauplius.
Results: The extract was found to contain polyphenols (34.60 %) and flavonoids (0.2536 %). It was effective against Staphylococcus aureus (125 µg.ml-1) and Escherichia coli (500 µg.ml-1) with moderate toxicity (CL50 = 893.62 μg.ml -1).
Conclusion: The study plant showed antimicrobial potential against S. aureus and E. coli, though further studies should be conducted to evaluate its toxicity and the possibility of its use to produce new drugs.

Key words: plant extracts, antimicrobial activity, toxicity.

RESUMEN

Introducción: Spondias mombin L. (Anacardiaceae) se utiliza en la medicina tradicional en la región semiárida de Brasil contra diversas enfermedades.
Objetivo: Evaluar el perfil fitoquímico y la actividad biológica del extracto etanólico de S. mombin.
Métodos: El extracto etanólico de la corteza se obtuvo mediante turbo-extracción y se sometió a ensayos fitoquímicos por espectrofotometría UV-VIS, la susceptibilidad microbiana se evaluó mediante microdilución y la toxicidad se determinó en nauplios de Artemia salina.
Resultados: Se encontraron polifenoles (34,60 %) y flavonoides (0,2536 %). El extracto fue eficaz contra Staphylococcus aureus (125 µg/mL-1 ) y Escherichia coli (500 µg/mL-1) con toxicidad moderada (CL50 = 893,62 μg/mL-1).
Conclusiones: La planta estudiada mostró potencial antimicrobiano contra S. aureus y E. coli, aunque deben realizarse más estudios para evaluar su toxicidad y la viabilidad para producir nuevos fármacos.

Palabras clave: extractos de plantas; actividad antimicrobiana; toxicidad.

 

 

Recibido: 11/09/2016
Aprobado: 28/06/2018

 

 

INTRODUCTION

Natural products are used for the treatment of disease since ancient times and are the only resources available for the treatment of different infections in several locations in the world. The increasing incidence of multidrug-resistant bacteria has attracted attention of scientific community for antimicrobial potential of natural products. Several tests reported the antimicrobial activity of different plant derived substances including their mechanisms of action.1

Spondias mombin L. (Anacardiaceae), popularly known as cajazeira, cajarana and cajazinho, is a fairly common tree species in northeastern Brazil. Ethnopharmacological studies of this plant reported use against diarrhea, conjunctivitis, digestive disorders, infections in general.2,3

Phytochemical screening showed the presence of phenols, tannins and proanthocyanins, saponins, anthraquinones, berberine, naphthoquinones, sesquiterpenes, indole and quinoline alkaloids and flavonoids in the plant4-6. These secondary metabolites have shown potential in pharmacological activities. Pharmacological studies of this plant have found antiepileptic, antipsychotic3, anthelmintic5, antiviral7 and anxiolytic8 activities.

So, in this study, it was carried out phytochemical studies and evaluation of biological activities of S. mombin extract.

 

 

METHODS

Bark of S. mombin Bark was collected at Fazenda Farinha, municipality of Pocinhos-PB at coordinates:7º07´54.53´´S e 36º07´14.51´´O. A voucher specimen (ACAM 000434) was deposited in the Arruda Câmara Herbarium of State University of Paraiba.

The plant material was dried in an air circulation oven at 40 °C and ground in a knife mill with a particle size of 10 mesh. The ethanol extract was obtained by turbo-extraction at room temperature (25 ± 2 ºC) using apparatus ultraturrax (Ika T-25) and 1L of ethanol a 96 %/100 g of plant, for 15 minutes at 15,000 rpm. After extraction, the solvent was removed on a rotary evaporator (Quimis) at 40 ° C and the sample diluted in dimethylsulfoxide (DMSO) at 10 %.

The content of total polyphenols and flavonoids of the extract was measured by spectroscopy in the visible region as described by Chaves et al.9

In microbiological tests were used standard strains American Type Culture Collection (ATCC) of pathogenic microorganisms Staphylococcus aureus (25923), Escherichia coli (25922), Pseudomonas aeruginosa (27853), Klebisiella pneumoniae (4352), Streptococcus mutans (25175), S. oralis (10557), S. salivarius (7073),Candida albicans (10231), C. guilliermondii (6260) e C. krusei (34135).

The minimum inhibitory concentration (MIC) was determined by a microdilution method in 96-well plates using Mueller-Hinton broth (Himedia) for all bacteria except for Streptococcus genus, for which was used BHI broth. Saboraud-dextrose broth was used for fungal strains. The plates were incubated at 37 ± 1 °C for 24 hours for bacteria and 25 ± 1 °C for fungi. Microbial growth was indicated by addition of 20 μL of 0.01 % aqueous resazurin solution (Sigma-Aldrich) with incubation at 37 ± 1 °C for 2 h. MIC values were identified as the lowest concentration in which no bacterial growth is visible. The assays were performed in triplicate.

For the toxicological test, were used Artemia salina nauplii, based on the technique described by Meyer et al.10 with minor adaptations. The nauplii hatched tank with synthetic sea water (pH = 8), ambient temperature and artificial lighting. Later they were separated into groups of 10 individuals, where except the control group, each received the solutions of the samples tested at different concentrations (2 000, 1 500, 1 000, 500, 250 μg.mL-1). The groups were subjected to artificial lighting and live larvae were counted after 24 hours. The experiment was performed in triplicate, and the values obtained, we estimated the LC50 through analysis Probit method, with 95 % confidence interval, using the program EPA Probit Analysis Program, version 1.5.

 

 

RESULTS

Phytochemicals tests revealed the polyphenols and flavonoids concentrations of the order of 34.60 % and 0.2536 %, respectively (Table 1).

 

In the analysis of antimicrobial activity, there was significant minimum inhibitory concentration of the extract against S. aureus (250 µg mL-1) and E. coli (500 µg mL-1), being the MIC over other strains bigger than 1000 µg mL-1 Table 2. In bioassay with A. salina, the extract showed LC 50 = 893.62 μg mL-1.

 

 

DISCUSSION

The turbo-extraction is cited in the literature 11,12 as that extracts large quantities of active principles, with yields higher than other techniques. This was verified by our research group in other studies (unpublished data). According to Marques and Vigo,13 this method presents high yields due to the crushing plant drug combined with continuous homogenization of solvent and vegetable drug.

Ríos and Recio (2005) suggest that in the study on medicinal plants, the extract should be considered as possessing antimicrobial activity if the MIC is <1 000 µg mL-1. In this work the extract was active on S. aureus and E. coli. Similar results were obtained by Abo et al.14 against the same bacterial species.

The activity is displayed by the tested extract probably related to the presence of polyphenolic compounds such as tannins and flavonoids, which are seen as having such activity.15,16 These compounds can destabilize the membrane and the bacterial metabolism, making it impossible to survive.16,17

The brine shrimp bioassay is a simple method proposed for detect toxicity of natural product. After 24 hours of exposure to extract was observed moderate toxicity according Meyer et al.10

The results demonstrate the potential of S. mombin against infections caused by S. aureus and E. coli, as well as moderate toxicity against A. salina. However, further studies are needed to verify the therapeutic viability of this natural product.

 

 

REFERENCES

1. Kalemba D, Kunicka A. Antibacterial and antifungal properties of essential oils. Current Medicinal Chemistry. 2003;10(10),813-29.

2. Cartaxo SL, Souza MMA, Albuquerque UP. Medicinal plants with bioprospecting potential used in semi-arid northeastern Brazil. Journal of Ethnopharmacology. 2010;131(2):326-42.

3. Lucena RFP, Albuquerque UP, Monteiro JM, Almeida CFCBR, Florentino ATN, Ferraz JSF. Useful Plants of the Semi-Arid Northeastern Region of Brazil - A Look at their Conservation and Sustainable Use. Environmental Monitoring and Assessment, 2007;125(1-3):281-90.

4. Ayoka AO, Akomolafe RO, Iwalewa EO, Akanmu MA, Ukponmwan OE. Sedative, antiepileptic and antipsychotic effects of Spondias mombin L. (Anacardiaceae) in mice and rats. Journal of Ethnopharmacology. 2006;103(2):166-75.

5. Ademola IO, Fagbemi BO, Idowu SO. Anthelmintic activity of extracts ofSpondias mombin against gastrointestinal nematodes of sheep: studies in vitro and in vivo. Tropical Animal Health and Production. 2005;37(3):223-35.

6. Coates NJ, Gilpin ML, Gwynn MN, Lewis DE, Milner PH, Spear SR, Tyler JW. SB-202742, a novel beta-lactamase inhibitor isolated from Spondias mombin. Journal of Natural Products. 1994;57(5):654-7.

7. Corthout J, Pieters LA, Claeys M, Vanden Berghe DA, Viletinck AJ. Antiviral Caffeoyl Esters from Spondias mombin. Phytochemistry 1992;31(3):1979-81.

8. Ayoka AO, Akomolafe RO, Iwalewa EO, Ukponmwan OE. Studies on the anxiolytic effect of Spondias mombin L. (Anacardiaceae) extracts. Afr. J. Trad. CAM. 2005;2(2):153-65.

9. Chaves TP, Fernandes FHA, Santana CP, Santos JS, Medeiros FD, Felismino DC, Santos VL, Catão RMR, Coutinho HDM, Medeiros ACD. Evaluation of the Interaction between the Poincianella pyramidalis (Tul.) LP Queiroz Extract and Antimicrobials Using Biological and Analytical Models. PloS one. 2016;11(5), e0155532.

10. Meyer BN, Ferrigni NR, Putnam JE. Brine shrimp: a convenient general Bioassay for active plant constituents. Planta Medica, 1982;45(5):31-4.

11. Politi FA, De Mello JC, Migliato KF, Nepomuceno AL, Moreira RR, Pietro RC. Antimicrobial, Cytotoxic and Antioxidant Activities and Determination of the Total Tannin Content of Bark Extracts Endopleura uchi. International Journal of Molecular Science, 2011;12(4):2757-68.

12. Souza APTB, Barni ST, Ferreira RA, Couto AG. Desenvolvimento Tecnológico de Soluções Extrativas Hidroetanólicas das Flores de Calendula officinalis L. Empregando Planejamento Fatorial. Latin American Journal of Pharmacy, 2010;29(1):13-21.

13. Marques LC; Vigo CLS. Preparação e Padronização de Extratos Vegetais. In: Leite JPV. Fitoterapia: Bases Científicas e Tecnológicas. São Paulo: Atheneu, 2009.

14. Abo KA, Ogunleye VO, Ashidi JS. Antimicrobial Potential ofSpondias mombin, Croton zambesicus and Zygotritonia crocea. Phytotherapy Research. 1999;13:494-7.

15. Chung K, Wei C, Johnson MG. Are tannins a double-edged sword in biology and health? Trends in Food Science and Technology. 1998;9(4):168-75.

16. Cowan MM. Plant products as antimicrobial agents. Clinical Microbiology Reviews. 1999;12(4):564-82.

17. Cushnie TPT, Lamb AJ. Antimicrobial activity of flavonoids. International Journal of Antimicrobial Agents. 2005;26(5),343-56.

 

 

Conflicts of interest

The authors declare no conflicts interest.



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