ANTIMICROBIAL ANALYSIS AND PHYTOCHEMICAL OF Mangifera indica AND Carica papaya LEAVES

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ANTIMICROBIAL ANALYSIS AND PHYTOCHEMICAL OF Mangifera indica AND Carica papaya LEAVES

3.1 NON-DISPOSABLES 
Extraction tank, beakers, measuring cylinder, water bath, tests tubes rack, test tubes, sample pipette, calibration pipette, Bunsen burner, matchet, diesel bunder, separating funnels, spatula, cork borer, refrigerator, meter rule, pencil distiller, eccentric oven, incubator, autoclave, clamp stand, inoculating hoop, spirit lamp, masking tape.
3.1.1   Disposables: Whitman Filter paper, petridishes, face mask, cotton wool, Aluminum foil, filter paper, masking tape, pen, permanent marker, hand gloves, hand towel, injection bottles, stock bottles.
3.1.2   Reagents: 
1% Aqueous hydrochloric acid
10% Fenic chloride
Acetic anhydride
Dragendorff reagent
Concentrated sulphuric acid
Sodium bicarbonate
Concentrated hydrochloride acide
Bromine water
Magnesium metal
Ammonium hydroxide solution
Fehlin chloride A and B
Benzene solution
Chloroform solution
Methanol (redistilled)
Glacial acetic acid
3.1.3   Solvents:
Sterile distilled water
Ethanol
Chloroform
Methanol
Ethlacetate
N-hexane
3.1.4   Media – Nutrient broth 
3.1.5   Organisms:  Codes
Staplylococcus aurous              LIO
Escherichia coli                         LIO
LIO – locally isolated organism

3.1.6 Drugs: 
40 grams of Mangifera indica extract
40 grams of Carica papaya extract
1 gram of methanol, ethylaceta, chloroform and N-hexane fractions of Maifera indica and Carica papaya respectively.
Genterincin injection (80mg/2ml) (control).
3.2       Methods:
3.2.1   Collection of Mangifera indica (mango) and Carica papaya
The fresh leaves of mango and pawpaw were collected in February, 2014 at Uyo, in Uyo Local Government Area of Ikwa Ibom State of Nigeria. Identification Number for Mangifera indica: UUH, No: 3C, Carica papaya: UUH, No: 18A.
3.2.2   Processing of the leaves of Mangifera indica and Carica papaya
The leaves were reduced to smaller sizes using knife and then dried under the sun several days separately. These leaves were further comminuted. The fragments were then stored in a polythene bags free of moisture until required.
3.2.3   Extractions 
600g of the comminuted leaves of Mangifera indica and Carica papaya respectively were macerated separately in methanol (redistilled) 1.5L respectively for 72 hours with intermittent shaking, all in separate tank extract was filtered using cotton wool and the filtrate further filtered using whatma no.1 filter paper to have the methanolic filtrate respectively. The extract were then concentrated to a constant weight. The concentrated extracts were weight separately and stored at 4oC in well closed beakers for further analysis.
3.2.4   Phytochemical screening:
Phytochemical screening for secondary metabolites in the plant were performed using standard procedures as described by (Sofowora 1993, Trease and Evans, 2006) and Harborne 1998 as follows:

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ANTIMICROBIAL ANALYSIS AND PHYTOCHEMICAL OF Mangifera indica AND Carica papaya LEAVES