INTRODUCTION:

Medicinal plant is need of pharmaceutical industry. According to WHO (World Health Organization) over 4 billion people i.e., 80% of the world population use herbal medicine. There are 250,000 plant species are present in earth but approximately only 35,000 to 70,000 plant species are used as medicinal purposes. Murraya paniculata is Asian plant its native from India, china, Bangladesh etc. and its distributed in Australia, Southeastern USA and central America. Murraya is genus of flowering plant. Over 14 global species of murraya is present and only three species i.e.

Murraya konini, Murraya paniculata and Murraya exotica is present in India. Murraya paniculata [L.] jack, family: Rutacesea, Synonym: Chalcapaniculata, other name is Kamini and orange jessamin. Murraya paniculatais evergreen shrubby plan. Small white flowers with aromatic nature, small leaves and hard wood. Murray paniculata is medicinal plant used as a anti diarrhoeal, anti inflamentry, diseases of teeth and gum, analgesic, antimicrobial, anticancer, antidiabetic activity. The flowers of Murraya paniculata [L.] jack are small white in colourwith 2-3 cm long x 1 cm in wide. It have a 7-13 bunch of flower on one spring and each flower contain 5 buds. The flower extract of Murraya paniculata was show antibacterial activity in gram positive and gram negative bacteria. The agar diffusion method is used for antibacterial in different - different concentration i.e. 10%, 25%, and 50% and the zone of inhibition is determine.

MATERIAL AND METHODS:

Collection of plant material:

Fresh flowers of Murraya paniculata [L.] jack were collected and rinsed it properly with water, dried in naturally approximately24-27⁰C for a one month and convert into powder form

Preparation of extract:

Fresh flowers powder approximately 150grams of Murraya paniculata extracted with petroleum ether to remove fatty substance and coloury matters by Soxhlet extraction method, then powder material was sprat and dried in naturally for a one day. After the process of drying the powder was weighted and further extracted with ethanol 500ml in 100gm of powder into Soxhlet. Soxhlet apparatus which run for ten cycles in duration of each cycle was about 90 minutes at 30⁰C temperature should be maintained. Again the powder was sprat and dried in room temperature for a one day and further extracted with water for aqueous extraction with Soxhlet for 10 cycles in duration of each cycle was about 80 minutes. Both extract (ethanol and aqueous) was filtered and concentrated by the water bath 40⁰C until drying then kept in desiccator. After drying both extract was stored in sterile air tight containers for further analysis.

Phytochemical analysis:

The qualitative analysis for phytochemical constitute that are present in extract. For this the specific test were performed to evaluate the presence particular phytochemicals. The following tests performed for checking the availability of alkaloids, flavonoids, steroids, phenols, saponins, cardiac glycosides, tannins, terpenoids etc.

Test for Alkaloids:

1 Mayer’s test: 2-3ml of filtrate, few drops of dil. HCl and Mayer’s reagent was added then shake well. Yellow precipitate show presence of alkaloids.

2 Wagners test: 2ml of filtrate and 1% HCl add. Then add 6 drops of Wagner’s reagent. Brownish-red precipitate indicates presence of alkaloids.

3 Dragendroff test: 2-3ml of filtrate, few drops of dil. HCl and add few drops of Dragendroff’s reagent add and shake well. Orange-brown precipitate indicates presence of alkaloids.

Test for Flavonoids:

1 Lead acetate test: 2-3 drops of extract, few drops of lead acetate was formation of yellow precipitate indicate the presence of Flavonoids.

2 Shinoda test: Few ml of extract and add few fragment of magnesium ribbon and add HCl dropwise heated that crimson red, pink scarlet colour appears after few minutes indicating presence of Flavonoids.

Teat for Cardiac glycosides:

1. Kelle-Killiani test: 5 ml of extract add 1 ml of conc. H₂SO₄, 2ml of Glacial acetic acid and 1 drop of FeCl₃ solution was added. Appearance of brown ring shows the presence of cardiac glycoside.

Test for Carbohydrates and Glycosides:

1. Fehling test: Take 2ml of filtrate and 2-2ml of Fehling solution A and B was added and solution was heated. A brick red precipitate indicates the presence of glycoside.

Fixed oil and fats test:

1. Spot test: Small quantity of extract was pressed between two filter paper. Appearance of oil stain on the paper indicates the presence of fixed oil.

Test of Steroids:

1. Salkowaski test: take 2ml of extract, 2ml of chloroform and 2ml of conc. H₂SO₄was added. By shaking this solution chloroform layer turned red and acid layer showed greenish yellow fluorescence indicating presence of Steroids.

Saponins test

1. Foam test: 0.5ml of filtrate and add 5ml of distilled water and shake well and stay it for 5 minutes. Formation of foam shows presence of saponins.

Proteins and Amino acids test:

1. Ninhydrin test: 2 drops of ninhydrin solution and added 2ml of filtrate. Purple colour indicate the presence of amino acid.

Phenolic and tannins test:

1. FeCl3 test: Few drops of 5%F_eCl₃ solution add in extract, appearance of deep blue colour indicate presence of tannins

2. Lead acetate: Add lead acetate solution to the extract white precipitate appeared.

3. Gelatin test: 2ml of extract, 1% gelatin solution containing 10% sodium chloride was added to obtained white precipitate.

Quinones test:

2ml of extract add conc. H₂SO₄and shake well for 5 min. shows rad colour.

Antibacterial assay:

The antibacterial assay is performed by the agar well diffusion method. In this method the nutrient agar media was prepared by using peptone, agar, beef extract, sodium chloride in sterilized equipment. All the equipment should be sterilized for antibacterial assay. There are two extract in different-different concentrations are used in this assay. For the Ethanolic extract 10%, 25% and 50% of concentration are make and for the Water extract 10%, 25% and 50% of concentration are make. There are two bacteria are used gram positive and gram negative. For the Gram positiveBacillus cereus are used and for the Gram negative Klebsiella pneumoniae are used. For the standard drug are used. After the making dish then the entire 12 dish are stay in incubator for growth at 35⁰C temperature for 3-4 days. The zone of inhibition of bacteria growth around each well in measured and determined. Antibacterial activity was evaluated by measuring zone of inhibition by using Hi-media zone scale.

Antifungal screening:

The antifungal activity is performed by disc diffusion method against one pathogenic fungal. For this the fungal medial was prepared. In this method the fungal agar media was prepared by using peptone, agar, glucose, distilled water in sterilized equipment. Make a slant culture with deep a fungus (Trichophyton rubrum) and stay it in incubator for fungus growth at 25⁰C temperature for 24 hour. Temperature should be maintained. After that again prepare a fungus culture without agar and slant culture is deep in second culture and stay second culture in incubator for 24 hour at 25⁰C temperature. Prepared third culture for disc diffusion method and make Petridis for different extract Ethanolic and Aqueous extract. The complete dis are stay in incubator for maintaining growth temperature at 25⁰-27⁰C temperature. The diameter of zone of inhibition produced by the extracts was then compared with the standard antibiotic.

Result and Discussion:

The Phytochemical analysis of flower extract of Murraya paniculata gave the following Phytochemical that are discussed in the table 1.

Table 1: Phytochemical analysis of Murraya paniculata flower

Sr. No.	Constituents	Test	Ethanolic Extract	Aqueous Extract
1.	Alkaloids	Dragendroff’s test Wagners teat Mayers test	+ + +	+ + +
2.	Flavonoids	Lead Acetate test Shinoda test	+ +	+ -
3.	Cardic Glycoside	Keller-Killiani’s test	+	+
4.	Carbohydrates & Glycosides	Fehling test	+	+
5.	Fixed oil & Fats	Spot test	+	+
6.	Steroids	Salkowaski test	-	-
7.	Saponins	Foam test	-	+
8.	Protein & Amino test	Ninhydrin test	+	-
9.	Phenolics &Tannin	F_eCl₃ test Lead Acetate Gelatin test	- + +	- + +
10.	Quinones	Quinones	+	+

[Positive (+) means present and Negative (-) means absent]

Zone of Inhibition:

Antibacterial activity of Ethanolic and Aqueous extract of Murraya paniculata flowers shown zone of inhibition.

Fig 1: Antifungal activity evaluation

CONCLUSION:

The Murraya paniculata is belongs to the family Rutaceae. The flowers of Murraya paniculata was taken for antibacterial and antifungal activity. The exhaustive extraction of the plant material was done with pet. Ether, Ethanol and water. Extract were screened for the prescience of medicinal active Phytoconstituents. The Flower extract of Murraya paniculata shows antibacterial and antifungal activity.

REFERENCE:

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Received on 19.11.2019 Modified on 31.12.2019

Asian J. Res. Pharm. Sci. 2020; 10(1):17-20.

DOI: 10.5958/2231-5659.2020.00004.1

Sr. No.	Concentration In %	Ethanolic Extract Zone of inhibition Average (mm)	Aqueous Extract Zone of inhibition Average (mm)	Sulfamethoxazole Zone of inhibition Average (mm)
1.	10%	22mm	24mm	53.12mm
2.	25%	26mm	30mm	55.75mm
3.	50%	35mm	37mm	52.35mm

Sr. No.	Concentration In %	Ethanolic Extract Zone of inhibition (mm)	Aqueous Extract Zone of inhibition (mm)	Sulfamethoxazole Zone of inhibition (mm)
1.	10%	28mm	35mm	50mm
2.	25%	31mm	38mm	53mm
3.	50%	41mm	43mm	54mm