Phytochemical and Antibacterial Screening of the Leaf Extracts of Bombax buonopozense
K Abdu*, H Ammani
Department of Pure and Industrial Chemistry, Bayero University, Kano, Nigeria
*Corresponding author: K Abdu, Department of Pure and Industrial Chemistry, Bayero University, Kano, Nigeria. Tel: +23464666023; Fax: +23464661480; Email: kabdu.chm@buk.edu.ng
Received Date: 21 March, 2017; Accepted Date: 23 March, 2017; Published Date: 30 March, 2017
1. Abstract
Fresh powdered
leaves (200 g) of Bombax buonopozense
was percolated with 95 % ethanol
(1000 cm3) for one week. It was
decanted, filtered and evaporated on rota vapor (R110) at 40oC to obtained the crude ethanol extract (F1).
It was macerated sequentially with chloroform, ethyl acetate, acetone and water
to obtained the chloroform soluble fraction (F2), ethyl acetate soluble
fraction (F3), acetone soluble fraction (F4) and water soluble fraction (F5)
respectively. All the five fractions were subjected to phytochemical screening
to check the presence of some secondary metabolites, also antibacterial
screening was carried out against some clinical isolates. Presence of
alkaloids, flavonoids, phenolic glycoside, tannins, saponins and anthraquinones
were detected in all/ some of the tested fractions. Also a remarkable zones of
inhibition on some of the baterial isolates by some fractions were recorded.
2. Keywords: Antibacteria; Inhibition; Isolates;
Maceration; Phytochemical
1. Introduction
[9]. The immature fruits are prepared as an emollient
for skin, decoction of the young leaves is used as a warm bath for febrile
children. The grounded bark is taken by pregnant women to increase lactation;
the extract from the bark is drunk or applied on the head for dizziness. The
gum resin from the bark is pulverized, mixed with oil and used to manage skin
diseases [10].
Two hundred grams (200 g) of the powdered leaves was percolated with 1000 cm3 of absolute ethanol for one week [11]. The extract was decanted, filtered and evaporated using rotary evaporator (R110) at 40ºC to obtain the crude ethanol extract which was labeled as F1 and kept in a refrigerator at 0oC before used.
3.1. Preparation of Test Solution and Disc
Concentration
·
Concentration
of 60µg/disc: From the stock
solution, 0.5 cm3 was taken into a Bijour bottle containing the discs and labeled
60µg/disc.
·
Concentration
of 30µg/disc: 0.5 cm3 of
DMSO was added into the remaining 0.5 cm3 of
the stock solution making 1 cm3. It
was divided into two and half of it (0.5 cm3) was
taken into another Bijour bottle containing the discs and was labeled 30µg/disc.
·
Concentration
of 15 µg/disc: to the remaining 0.5 cm3 of 30
µg/disc solution, 0.5 cm3 DMSO
was added to make it 1 cm3. It was also divided into two
and half (0.5 cm3) of it was taken into another Bijour bottle containing the discs
and labeled 15 µg/disc.
Phytochemical
screening was carried out which involves the tests for alkaloids, flavonoids,
phenolic glycosides, anthraquinones, tannins and saponins. The crude ethanol
extract (F1), acetone fraction (F4) and the aqueous fraction (F5) were found to
contain all the tested secondary metabolite. The chloroform fraction (F2) also
contains all the tested secondary metabolites except phenolic glycosides. While
the ethyl acetate fraction (F3) was found to contains only phenolic glycosides,
anthraquinones and alkaloids. All the fractions (F1-F5) were found to form an
orange red precipitate/turbidity when treated with Draggendoff’s reagent which
indicates the presence of alkaloids [15].
Similarly, all the fractions give a reddish-pink colour when treated with
ammonia which confirmed the presence of anthraquinones in all the fractions [12].
Fractions F1, F2, F4 and F5 show the appearance of a red colour when treated
with conc. HCl which indicates the presence of flavonoids [12]. Similarly,
fractions F1, F2, F4 and F5 gives a dark black coloured precipitate with ferric
chloride which turned green-black to blue-black colouration on dilution which
indicates the presence of tannins [13]. Also
fractions F1, F2, F4 and F5 shows a persistent frothing when shaken with
distilled water which indicates the presence of saponins [12].
Fractions F1, F3, F4 and F5 were found to show a red
colouration which disappeared upon addition of distilled water which confirmed
the presence of phenolic glycoside. The results were shown in Table
2 below.
The antimicrobial activity
of the different fractions of B. buonopozenes was
investigated against some pathogens. Some fractions were found to possess the
bioactivity against some of the tested microorganisms. The tested organisms
include Klebsiella spp., Pseudomonas
auriginosa, Proteus spp., E. coli and Staphylococcus aureus. The crude ethanol extract
exhibited the highest activity on all the tested pathogens at all
concentrations, but with highest inhibition on E. coli and Klebsiella spp. both at the
concentration of 60µgml-1. It was followed by chloroform fraction which
showed remarkable zones of inhibition on E. coli, Klebsiella spp., Pseudomonas auriginosa and Proteus spp. but
with highest inhibition on E. coli and Pseudomonas auriginosa at concentration of 60µg ml-1.
The fraction with least activities was found to be aqueous fraction which
exhibited low activity on Kebsiella spp., Pseudomonas auriginosa and
E. coli (Table 3).
Fraction |
Texture |
Colour |
Weight (g) |
Crude ethanol extract (F1) |
Gummy |
Black |
37.39 |
Choloroform fraction (F2) |
Gummy |
Black |
3.07 |
Ethyl acetate fraction (F3) |
Gummy |
Black |
2.09 |
Acetone fraction (F4) |
Gummy |
Dark brown |
1.58 |
Aqueous fraction (F5) |
Gummy |
Brown |
10.8 |
Table 1: Extraction results with the physical properties of the extracts.
Secondary Metabolites |
F1 |
F2 |
F3 |
F4 |
F5 |
Phenolic gylcosides |
+ |
- |
+ |
+ |
+ |
Anthraquinones |
+ |
+ |
+ |
+ |
+
|
Flavonoids |
+ |
+ |
- |
+ |
+ |
Tannins |
+ |
+ |
- |
+ |
+ |
Saponins |
+ |
+ |
- |
+ |
+ |
Alkaloids |
+ |
+ |
+ |
+ |
+ |
Keys: ‘+’ indicate presence, ‘-‘indicate absence. |
Table 2: Results of the phytochemical screening.
Zones of Inhibition (mm) at different concentrations in µg/mL |
|
|||||||||||||||||
Pathogens
|
F1 |
F2 |
F3 |
F4 |
F5 |
AUG |
||||||||||||
60 |
30 |
15 |
60 |
30 |
15 |
60 |
30 |
15 |
60 |
30 |
15 |
60 |
30 |
15 |
(30 µg) |
|||
Klebsiella spp. |
12 |
10 |
8 |
9 |
8 |
6 |
9 |
7 |
- |
- |
- |
- |
8 |
7 |
6 |
23 |
||
Pseudomonas auriginosa |
10 |
9 |
9 |
10 |
9 |
7 |
8 |
7 |
6 |
8 |
6 |
- |
8 - |
7 |
6 |
30 |
||
Proteus spp. |
9 |
8 |
7 |
7 |
6 |
- |
- |
- |
- |
10 |
8 |
7 |
|
- |
- |
16 |
||
E. coli |
13 |
11 |
10 |
12 |
10 |
9 |
9 |
8 |
7 |
10 |
8 |
7 |
9 |
7 |
- |
33 |
||
Staphylococci. aureus |
9 |
7 |
6 |
- |
- |
- |
7 |
6 |
- |
8 |
6 |
- |
- |
- |
- |
27 |
||
Keys: AUG = Augmentin (standard) |
Table 3: Antibacterial screening results.
- Mann A, Gbate M, Nda-Umar A (2003) Medicinal and Economic plants of Nupe land. Jube-Evans Books and publications, Bida Nigeria 35-36.
- Beentje H, Sara S (2001) Plant Systematic and Phytogeography for the understanding of African Biodiversity, Systematic and Geography of plants 71: 284-286.
- Dubost G, Gabon J (1984) Comparison of Diets of frugivorous forest mammals 65: 298-316.
- Addo-Fordjour P, Obeng S, Addo MG, Akyeampong S (2009) Effects of human disturbances and plant invasion on Liana community structure and relationship with trees in the Tinte Bepo Forest Reserve, Ghana. Fonnr Manage 258: 728-734.
- Gyasi E. Agyepong G. Ardayfio-Schandorf E, Enu-Kwesi L, Nabila J, et al. (1995) Production pressure and environmental change in the forest-savanna zone of Southern Ghana. Glob Environ Chang 5: 355-366.
- Kamanzi, Atindehou K, Schmid C, Brun R, Koné MW, et al. (2004) Antitrypanosomal and antiplasmodial activity of medicinal plants from Côte d’ivoire. J Ethnopharmacol 90: 221-227.
- Asase A, Oppong-Mensah G (2009) Traditional antimalarial phytotherapy remedies in herbal markets in Southern Ghana. J Ethnopharmacol 126: 492-499.
- Madge C (1998) Therapeutic landscapes of the Jola, the Gambia, and West Africa. Health Place 4: 293-311.
- Irvine FR (1961) Woody plants of Ghana (1). Oxford University Publishers, London 897-955.
- Insoll T (2007) ‘Natural’ or ‘human’ spaces? Tallensi sacred groves and shrines and their potential implications for aspects of northern European prehistory and phenomenological interpretation. Nor Archaeol 40: 138.
- Abdu K, Echeme JO (2006) Phytochemical and Antimicrobial Screening of the Ethanolic Extract of the Leaves of Anogeissus leiocarpus. BEST 3: 162-164.
- Sofowora A (1993) Medicinal plant and traditional medicine in Africa. Spectrum Books ltd (Ibadan) 142-144.
- Harbone JB (1998) Phytochemical Methods, Chapman and Halls: London 91.
- National Committee for Clinical Laboratory Standards (2008) Performance Standards for Antimicrobial Testing, Wayne PA.
- Evans WC (2002) Trease and Evans Pharmacognosy, (15th edn). Bailliere Tindall: London 253: 289- 333.
- Alma MH, Mavi A, Yildirim A, Digrak M, Hirata T (2003) Screening chemical composition and in vitro antioxidants and antimicrobial activities of the essential oils from the Origanum syriacum L. growing in Turkey. Biol Pharm Bull 26: 1735-1739.
- El Astal ZY, Aera A, Aam A (2005) Antimicrobial activity of some medicinal plant extracts in Palestiine. Pak J Med Sci 21: 187.
- Godwin Akuodor C, Augustine Essien D, Jemilat Ibrahim A, Augustine Bassey, Joseph Akpan L, et al. (2011) Phytochemical and antimicrobial properties of the methanolic extracts of Bombax buonopozense leaf and root, As J Med Sci 2: 190-194.
- Mann A, Salawu FB, Abdulrauf I (2011) Antimicrobial activity of Bombax buonopozense. Beav. (Bombacaceae) edible floral extracts. Eur J Sci 48: 627-630.
- Gupta, Oliver-Bever B (1983) Medicinal plants in tropical West Africa III. Anti-infection therapy with higher plants. J Ethnopharmacol 9: 1-83.