INTRODUCTION:

Ixora coccinea:

Botanical Name	*Ixora coccinea*
Common Name	Bakora (Marathi local Name)
Classification	Kingdom	Plantae
	Subkingdom	Tracheobionta
	Division	Magnoliophyta
	Class	Magnoliopsida
	Subclass	Asteridae
	Order	Gentianales
	Family	Rubiaceae
	Genus	Ixora
	Species	Coccinea

Ixora is a sort of blossoming plants in the Rubiaceae family. Red Ixora blossoms are normally utilized in Indian people medication. Ixora coccinea is a local to tropical South - East Asia, including Southern India, Sri Lanka and so on. The regular names of Ixora coccinea are west Indian Jasmine, Rangan, Kheme, Ponna, Techi, and Jungle geranium. Ixora plants by and large incline toward acidic soil for their growth. In tropical atmospheres this plant blooms year around and its fruits are utilized as a dietary source. Ixora coccinea is a thick, multi-spread evergreen bush, attractive up to 12 ft in height as shown in Figure 1. It has an adjusted structure, leaves are around up to four-inch-long, with whole edges, and are conveyed in inverse sets or whorled on the stems. The blossom of Ixora coccinea plant is traditionally used as antioxidant^1,6,8,9,19, anti-inflammatory⁶, antipyretic drug, digestive and constipating etc^3,4,5. The Stems of Ixora coccinea plants are used as astringent, sedative, stomachic, febrifuge and antiseptic¹³ and also diverse pharmacological properties^12,17 including anti-inflammatory, antimitotic activities¹⁴ etc^7,11. The synthesis of metal oxide and metal oxide nanocomposites¹⁵ from ixora coccinea plants extract²¹ are also employed. The major phytochemical constituents^2,4,9,10 present in Ixora coccinea such as flavonoids, tannins, Coumarin, Anthocyanin, terpenoids, Alkaloid, Phenol, Saponin, Anthraquinone and Amino acid¹⁶, till todays 54 compounds have been reported. Anticancer activity of the leaves of Ixora coccinea also reported due to the presence of alkaloid and camptothecin.

MATERIAL AND METHODS:

Isolation, Growth and Identification:

In the clinical investigation the parasitic fungi were segregated from scalp of individual experiencing contagious contamination and isolated¹⁸ on sabouraud's media slants and after incubation culture was kept at 4 °C in cooler. The isolated culture was screened by plating the scalp swab on sabouraud's media enhanced with 2% lipid source such as olive oil. The creature was distinguished based on cultural, microscopic and biochemical strategies as shown in Figure 2.

Collection of the plant material:

The leaves, blossoms and stem parts of an Ixora coccinea was collected from Nashik, Maharashtra (India).

Preparation of the Ixora coccinea plant extract:

The plant parts were washed several times with distilled water to remove the contaminated particles. They were shade dried, powdered are shown in figure 3 and subjected to successive 90% ethanol extraction by Overnight extraction method as shown in figure 4. Leaf, blossoms and stem part of an Ixora coccinea plants were extracted in 1: 10 ratios. Extracts were concentrated, dried and stored for further antifungal analysis are shown in Figure 5.

Synthesis of Ag nanoparticles from the Ixora coccinea plant:

The Ag nanoparticles were biosynthesized by using an aqueous extract of an Ixora coccinea and recovered the Ag nanoparticles by ultra-centrifugation for characterization. The 100ml of fresh leaf extract was added into the aqueous solution of 1 mM²⁰ Ag (NO₃) (50 ml) as shown in Figure 6, After addition, tubes were incubated in a dark condition for 1 days. Ag nanoparticles synthesis was confirmed by UV spectrophotometer, solution was centrifuged up to 25000 rpm for 10 minutes and dried under hot oven. The dried Ag nanoparticles were re-dissolved in sterile distilled water and further used for antifungal activity.

Antifungal activity of the plant extracts and Ag nanoparticles against Malassezia furfur by well diffusion method.

50 µl of the samples were loaded on the wells of sabouraud's dextrose media with 2% olive oil. Plates were incubated for 2-4 days and after incubation zone of inhibition as shown in table 1 and 2, was measured and compared with positive and negative control are shown in figure 7 and 8.

RESULTS:

Isolation and Identification of Malassezia furfur:

Figure 2 Isolation & Microscopic image of Malassezia furfur.

Malassezia furfur grew well in Sabouraud’s dextrose media enriched with 2% olive oil. Malassezia furfur grew as white to tan cream colored colonies smooth pasty consistency on Sabouraud’s media and the cells appeared bottling shaped when observed microscopically.

Extraction of Ixora coccinea plant parts in powder form:

Figure 3 Leaf, blossom and Stem powder

Overnight Extraction method:

Figure 4 Ethanolic extracts of Blossom, leaf and Stem of Ixora coccinea plants

Ixora coccinea leaf, blossom and stem extracts:

Figure 5 Dried Ixora coccinea ethanolic leaf, stem, and blossom extract

Synthesis of the Ag nanoparticle from the Ixora coccinea plant extract.

Figure 6 Synthesis of the Ag nanoparticle from the Ixora coccinea plat extract.

The formation of Ag nanoparticle by the aqueous extract of Ixora coccinea plant (Leaf (L), Stem (S) and Blossom (B)) in the solution of 1mM²⁰ Ag (NO₃) was indicated by change in color. This was confirmed by UV spectrophotometry with λmax = 434 nm.

Antifungal activity of the plant extracts against Malassezia furfur by well diffusion method:

Figure 7 Antifungal activity of the ethanolic plant extracts (A) L-Leaf, S–Stem, B–Blossom and negative control, (B) Positive control

Table 1 Zone of inhibition of ethanolic plant extracts

Sr. No	Ethanolic plant extraction parts	Zone of inhibition in mm
1	Leaf extract	7.6
2	Stem extract	4.33
3	Blossom extract	7.3
4	Positive control	7.33

Antifungal activity of the Ag nanoparticles against Malassezia furfur by well diffusion method:

Figure 8 Antifungal activity of silver nanoparticles (A) positive control (B) Ag nanoparticle 4:2 ratio of leaf (L), stem (S) and blossom (B).

Table 2 Zone of inhibition of biosynthesized silver nanoparticles from Ixora coccinea plant in 1:5 and 4:2 ratio

Silver nanoparticle synthesized plant part	Zone of inhibition (mm) 4:2
Leaf	7.95
Stem	5.42
Blossom	7.40

DISCUSSION:

Our study had shown that Ixora coccinea plant had an antifungal activity against Malassezia furfur. Plants used in traditional medicine are assumed to be safe due to the long-term use by traditional healers. Information about the safety and effective use of medicinal plants is difficult to find due to lack of rigorous clinical studies and limited toxicological data available. Preliminary qualitative phytochemical screening gives a clue for the medicinal aptitude of the herb. In the conducted study, bioactive compounds that impart biologically active nature to the plant were screened and results ensured the presence of flavonoid, terpenoid, coumarin, tannin, phenol, alkaloid, saponin, anthocyanin, anthraquinone and amino acids. The presence of wide range of phytochemical constituents indicates that the plant could be used in a multitude of ways which may be beneficiary to the population. An important part of natural products from plants, biomolecule and secondary metabolites usually exhibits some kind of biological activities. The usefulness of plant materials medicinally is due to the presence of bioactive constituents such as alkaloids, flavonoids, tannin and phenolic compound. The plant active constituent analysis is very important step as it gives the information regarding presence or absence of particular primary and secondary metabolites in the extract of various parts of plant. Blossom extract had shown higher concentration of flavonoid as compared to leaf extract. Natural anti-oxidants have attained worth reputation in treating several diseases and have raised the value of folk herbal medicines in the modern era. The bio reduction of Ag+ in the aqueous extract was monitored by sampling the reaction mixture by using UV-vis spectroscopy. Addition of the aqueous leaf extract of Ixora coccinea to 1mM solution of Ag (NO₃) led to the appearance of yellow color as result of formation of Ag nanoparticles in the solution. Before reaction, the Ag containing solution is colorless but changes to a brownish color on completion of the reaction. The brown color of the medium could be due to the excitation of surface plasmon vibrations, typical of the Ag nanoparticles. The UV-Vis spectra recorded from the aqueous Ag (NO₃) Ixora coccinea leaf broth. A strong characteristic absorbance peak at around 434 nm was observed. The UV-Vis absorption spectrum recorded for the solution shows the characteristic surface plasmon resonance band for silver nanoparticles in the range of 400 - 440 nm. Antifungal activity of silver nano particles was carried out, zone of inhibition in the plate showed that Ag nanoparticles synthesized using aqueous leaf extract of Ixora coccinea have the antifungal activity against Malassezia furfur through the inhibition zone formation. Zone of inhibition was measured and compared with control Ag (NO₃) solution. On comparison with the Ag (NO₃) and plant extracts Ag nanoparticles out performed in the antifungal effect. The results of the investigation showed that Ag nanoparticles synthesized from Ixora coccinea leaf and blossom extracts possess discrete antifungal activity against clinically isolated pathogenic Malassezia furfur. In our study, Ixora coccinea plant extract had shown an effective and good antifungal activity. The world market is also moving towards herbal medicines for health care, health foods and for cosmetic purposes including hair preparations. Ixora coccinea plant extracts were showing good antifungal activity, Leaf and blossom extract of the Ixora coccinea plant had more antifungal activity and this could be because of their active compound like flavonoid. The present study gives significant information about the higher antifungal activity of Ixora coccinea at low concentration which can be exploited for polyherbal preparation.

CONCLUSION:

This research work can be successfully concluded by the biosynthesis of Ag nanoparticles from Ixora coccinea plant and compared the antifungal property of ethanolic extract of plant and synthesized silver nanoparticles. Silver nanoparticles were synthesized and confirmed by spectrophotometry, a strong characteristic absorbance peak at around 434 nm was observed. Effective antifungal activity was shown by plant extract as well as the synthesized green silver nanoparticles. When compared, antifungal activity of biosynthesized Ag nanoparticle is more effective than the plant extracts. We can use this nanoparticle for the formulation of natural and eco-friendly products.

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Received on 29.03.2020 Modified on 13.04.2020

Asian J. Research Chem. 2020; 13(3): 198-202.

DOI: 10.5958/0974-4150.2020.00038.3