ANTIMICROBIAL ACTIVITY OF GARCINA KOLA EXTRACT ON WOUND SAMPLE

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ANTIMICROBIAL ACTIVITY OF GARCINA KOLA EXTRACT ON WOUND SAMPLE

ABSTRACT

Garcinia kola Heckel (Guttiferae) seed, has found use in folkloric medicine of Southern Nigeria for the treatment of toothache and prevention of dental caries. The crude ethanolic extract, chromatographic fractions and isolated constituents of Garcinia kola seed against clinical strains of dental-caries-causing and related microorganisms is being evaluated. Antimicrobial evaluations were done by testing different concentrations of the crude extract, vacuum liquid chromatographic (VLC) fractions and pure isolates against Streptococcus mutans, Streptococcus viridans and Staphylococcus aureus in already set blood agar with gentamicin as the reference standard. The zones of inhibition and minimum inhibitory concentrations (MIC) were determined as appropriate. Fraction N, eluted with (hexane: ethyl acetate 70: 30), exhibited the highest activity with MIC’s of 1.50 mgml ¹ and 0.33 mgml ¹ while the pure isolates 1 (cycloartenol) and 2 (24-methylenecycloartanol) gave MIC’s of 0.17 mgml ¹ and 0.38 mgml ¹ against Streptococcus mutans and Streptococcus viridans respectively. Isolate 3 (garcinianin) gave MIC of 1.0 mgml ¹ against Streptococcus mutans but there was no significant activity against Streptococcus viridans and Staphylococcus aureus. The results provide justifications for the folkloric use of Garcinia kola Heckel (Guttiferae) for dental caries-related health problems while the isolated compounds may also serve as templates for future antimicrobial drug development.

CHAPTER ONE

INTRODUCTION

BACKGROUND OF THE STUDY

he kola nut is the fruit of the kola tree, a genus (Cola) of trees that are native to the tropical rainforests of Africa. The caffeine-containing fruit of the tree is used as a flavoring ingredient in beverages, and is the origin of the term “cola”. The kola nut is a caffeine-containing nut of evergreen trees of the genus Cola, primarily of the species Cola acuminata and Cola nitida. Cola acuminata, an evergreen tree about 20 metres in height, has long, ovoid leaves pointed at both the ends with a leathery texture. The trees have yellow flowers with purple spots, and star-shaped fruit. Inside the fruit, about a dozen round or square seeds develop in a white seed-shell. The nut’s aroma is sweet and rose-like. The first taste is bitter, but it sweetens upon chewing. The nut can be boiled to extract the caffeine. This tree reaches 25 meters in height and is propagated through seeds. C. nitida and C. acuminata can easily be interchanged with other Cola species.

Garcinia kola Heckel (Guttiferae) seed has been used extensively in traditional medicine for the treatment of various diseases (Taiwo et al., 1999). The seeds are chewed in Southern Nigeria and parts of West Africa as a masticatory and are also used to prevent or relieve colic, headaches, chest colds and cough. The seeds are also chewed as an aphrodisiac and find use in the treatment of dysentery, bronchitis and throat infections (Braide, 1990). The stem bark is used as a purgative and for the treatment of malignant tumors. The sap is used for parasitic skin disease while the latex (gum) is used internally for gonorrhea treatment. It is applied externally to treat fresh wounds (Tona et al., 1999). The plant has also been employed in the treatment of liver disorders while the fruit yield favorite bitter chewing sticks sold in small bundles in local markets across West Africa (Iwu, 1993).

In the present study, a bioactivity-monitored fractionation and isolation of antimicrobial constituents of Garcinia kola Heckel seed with activity against dental Thin layer chromatographic analysis of the extracts on silica gel plates (0.25 mm thickness) with appropriate solvent systems and chromogenic reagents was also carried out to detect the secondary metabolites present.

Isolation of compounds: The crude ethanolic extract was chromatographed on silica gel (Vacuum Liquid Chromatographic) (VLC) and eluted with solvents of increasing polarity (hexane- methanol gradients), followed by monitoring with thin layer chromatography. Similar fractions were pooled together and labeled K, M, N, O and P. Fraction N (11.68 g) from the VLC fractionation was chromatographed on silica gel (column chromatography) and on further purification with preparative TLC (toluene: ethyl acetate: acetone 8:2:1) yielded compounds 1, 2 and 3. The pure isolates were subsequently characterized using spectroscopic means (Infra Red, Nuclear Magnetic Resonance, and Mass Spectroscopy).

Antimicrobial screening of crude extracts and chromatographic fractions: Different concentrations of the crude extract ranging from 400 mgmlˉ¹ to 25 mgmlˉ¹ was prepared and was used to fill the bored wells made in already set blood agar containing Staphylococcus aureus, Streptococcus mutans and Streptococcus viridans in different Petri- dishes and incubated at 37 ⁰C for 24 hours. The zones of inhibition were measured while gentamicin (2.5 µgml⁻¹) was used as positive control as shown in Table 1.

1.2 STATEMENT OF THE PROBLEM

Previous studies have revealed that Garcinia kola Heckel contains a mixture of phenolic compounds including xanthones and benzophenones (Locksley et al., 1971; Hussain et al., 1982). Flavonoid derivatives including apigenin, fisetin and biflavonoids such as ametoflavone have also been isolated from Garcinia kola Heckel seeds (Iwu et al., 1982). Kolaviron, an extractive from the seed has also been shown to possess remarkable prophylactic and therapeutic action on damaged stomach (Ibironke et al, 1997; Olaleye, 2005), liver (Farombi, 2000; Iwu, 2001; Adaramoye et al., 2005;), colon (Ige et al, 2010; Farombi et al, 2013) and brain (Ijomone et al, 2012)

1.3 OBJECTIVES OF THE STUDY

1.To understand the antimicrobial activity of garcina kola extract on wound sample

2. To understand the relationship between the antimicrobial activity of garcina kola and the solution to wound

1.4 RESEARCH QUESTIONS

1.What is the antimicrobial activity of garcina kola extract on wound sample

2. What is the relationship between the antimicrobial activity of garcina kola and the solution to wound

RESEARCH HYPOTHESIS

H0: There is no relationship between the antimicrobial activity of garcina kola and the solution to wound

H1: There is a relationship between the antimicrobial activity of garcina kola and the solution to wound

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