INTRODUCTION:

Psidium guajava L. (Myrtaceae), an evergreen small tree, commonly known as guava in English, is an important food crop and medicinal plant indigenous to South America and grown in tropical and subtropical countries. It is widely used as food and in folk medicine around of the world¹. It is commercially cultivated in India for its consumable and palatable fruits. A number of metabolites in good yield and some have been shown to possess useful biological activities belonging mainly to phenolic compounds, flavonoids, carotenoids, terpenoids and triterpenes.

Extracts and secondary metabolites of this plant, particularly those from leaves and fruits possess useful pharmacological activities². P. guajava is mainly known for its antispasmodic and antimicrobial properties in the treatment of diarrhoea and dysentery³. It has also been used extensively as an oral hypoglycaemic agent. Several pharmacological studies have demonstrated the ability of this plant to exhibit antioxidant, hepatoprotection, anti-allergic, antimicrobial, antigenotoxic, antiplasmodial, cytotoxic, antispasmodic, cardioactive, antitussive, antidiabetic, anti-inflammatory and anti-nociceptive activities, supporting its traditional⁴. Natural products have contributed significantly towards the development of modern⁵. Recently traditional medicine worldwide is being reevaluated by extensive research on different plant species and their active therapeutic principles. The rich wealth of plant kingdom can represent a novel source of newer compounds with significant antiinflammatory activities. The major merits of herbal medicine seem to be their perceived efficacy, low incidence of serious adverse effects and low cost⁶.

MATERIALS AND METHODS:

Plant material:

Psidium guajava leaves has been collected from Karad, Maharashtra, India. Department of Botony, Yashwantrao Chavan College of Science, Karad has identified the plant and authenticated it.

Preparation of Psidium guajava leaf powder:

Fresh guava leaves were collected and air dried for 10 days. The dried leaves were then crushed into a blender and churned to form a coarse powder. The powder was collected in an air-tight jar, and stored away from sunlight in a cool, dry place.

Preparation of plant extract:

Extraction of malvastrum coromandelianum was done by microwave extraction method further filtered and excess solvent present was evaporated and dried extract were collected and subjected for activity studies.

Chemicals:

Chemicals have been purchased from the Research lab, Mumbai, India, and the solvent has been purified by distillation. DMSO as solvent, Diclofenac sodium as a standard, 50 μg/ml, 100 μg/ml were concentration. UV spectrophotometer instrument for absorbance for anti-inflammatory activity and fluconazole as a standard drug used for antifungal activity.

Anti-inflammatory activity:

Method: In vitro anti-inflammatory activity by protein denaturation method.

The mixture (10 ml) consisted of 0.4 ml of egg albumin (from fresh hen’s egg), 5.6 ml of phosphate buffered solution (PBS, pH 6.4) and 4 ml of varying concentration of test samples so that final concentration become 50 μg/ml,100 μg/ml. Similar volume of DMSO served as control. Then the mixtures were incubated at (37°C ± 2) for 15 min. and then heated at 70°C for 5min. After cooling, their absorbance was measured at 660 nm (JASCO UV Spectrophotometer) by using vehicle as blank and their viscosity was determined by using Ostwald viscometer. Diclofenac sodium at the final concentration of 50 μg/ml, 100 μg/ml were used as reference drug and treated similarly for determination of absorbance and viscosity and % inhibition of protein denaturation were calculated [7-9].

Antifungal activity:

Preparation of Saboraud’s broth slants and sub-culturing of microorganisms:

Saboraud's agar inclinations was set up by utilizing Agar 500mg, 250 mg of peptone, Glucose 500 mg was broken down in 50ml of refined water, bubbled and afterward poured it in the test tube and the test cylinders was stopped with cotton and after that disinfected in autoclave at 15 lbs weight (121⁰C) for 15 min. After the cleansing the cylinders containing the Saboraud's agar were kept in slanted position for ½ hrs. At that point on the strong surface of these inclinations the unadulterated culture of the test growths for example Candida Albicans, Aspergillus Niger was streaked in aseptic condition and after that brooded at 37⁰C for 24 hrs.

Preparation of culture media for Antifungal sensitivity test:

Saboraud's agar pH (5.4) media was set up by dissolving Agar 20gm, Glucose 20gm, and Peptone 10gmin 1000 ml bubbling refined water and blended well. At that point it was sanitized in autoclave at 15 lbs weight (121⁰C) for 15mins. After sanitization the media was cooled up to 45⁰C and after that it was poured in sterile Petri plates in aseptic condition. Around 20-25 ml of media was poured in each plate. At that point the media from the plate was permitted to get hardened at room temperature.

Inoculation of suspension of fungi on culture media:

Sterile, non-dangerous cotton swab was dunked in to the institutionalized inoculums (turbidity as balanced as to acquired intersecting development on the Petri plate) and afterward the whole agar surface of the plate was streaked with the swab multiple times, turning the plate at 60^o point between streaking. After that the streaked inoculums were permitted to dry for 5-15mins with cover set up. At that point bore was punched on the readied plates by utilizing clean well (8mm). The 100μl portion of combined nanoparticles arrangement and standard medication Fluconazole was stacked per bore separately in aseptic condition with assistance of clean micropipette. Plates was kept at room temperature for 30 min and afterward hatched at 37⁰C for 24 hrs. The width of the zones of inhibition was estimated with the assistance of scale in mm¹⁰.

RESULTS AND DISCUSSION:

Anti-inflammatory activity:

Table No. 1 In vitro anti-inflammatory activity of extract measuring the percentage inhibition

Sr. no	Test subs	% inhibition of protein denaturation		Viscosity(cp)
Sr. no	Test subs	50µg/ml	100µg/ml	50µg/ml	100µg/ml
1	Methanolic extract	71.45	74.17	2.42	3.82
2	Ethyl acetate extract	75.25	77.48	2.91	3.76
3	Standard (diclofenac)	78.12	80.35	2.74	3.92

Antifungal activity:

The antifungal assay were performed and results were is shown in (Table no. 2). Psidium guajava ethyl acetate extract showed significant activity against against C. albicans and A. niger

Table No. 2 Antifungal screening of Psidium guajava extract

Test subs	Concentration	Zone of inhibition
Test subs	Concentration	*C. albicans*	*A. niger*
Methanolic extract	10 µg/ml	4 mm	6 mm
	25 µg/ml	8 mm	11 mm
	50 µg/ml	10 mm	13 mm
Ethyl acetate extract	10 µg/ml	5 mm	9 mm
	25 µg/ml	9 mm	10 mm
	50 µg/ml	11 mm	14 mm
Std.(Fluconazole)	10 µg/ml	6 mm	8 mm
	25 µg/ml	10 mm	12 mm
	50 µg/ml	14 mm	16 mm

CONCLUSION:

Denaturation of proteins is a well-documented cause of inflammation. Several anti-inflammatory drugs have shown dose dependent ability to inhibit thermally induced protein denaturation. It has been reported that one of the features of several non steroidal anti-inflammatory drugs is their ability to stabilize (prevent denaturation) heat treated albumin at the physiological pH. This anti-denaturation effect was further supported by the change in viscosities. It has been reported that the viscosities of protein solutions increase on denaturation. In the present study, the relatively high viscosity of control dispersion substantiated this fact. Ability of Psidium guajava to bring down thermal denaturation of protein is possibly a contributing factor for its antiinflammatory activity. By performing antifungal activity ethyl acetate extract showed significant activity against against C.albicans and A. niger.

ACKNOWLEDGEMENT:

I express my sincere thanks to Vice-Principal Prof. Dr. S. K. Mohite for providing me all necessary facilities and valuable guidance extended to me.

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Received on 02.06.2020 Accepted on 14.07.2020

Research J. Topical and Cosmetic Sci. 2020; 11(2):62-64.

DOI: 10.5958/2321-5844.2020.00011.4