SEM/EDAX AND ANTIMICROBIAL ANALYSIS OF …

SEM/EDAX AND ANTIMICROBIAL ANALYSIS

OF STACHYTARPHETA JAMAICENSIS IN SILVER NANO PARTICLES

G.ALDOUS JENIN1*, S.S.SAJITHA2, S.Muthumariappan3, P. METILDA4

1. Assistant Professor, Department of Biochemistry, Lekshmipuram College of Arts and Science, Neyyoor.

2. Assistant Professor, Department of Chemistry, Annai Velankanni College, Tholayavattam.

3. Assistant Professor, Department of Chemistry, V.O.Chidambaram College, Tuticorin.

4. Assistant Professor, Department of Chemistry, Nesamony Memorial Christian College, Marthandam

Affiliated to Manonmanium Sundaranar University, Tirunelveli. Tamilnadu, India.

Abstract:

The synthesis of nanoparticles from plant is rapid, low-cost, eco-friendly and a single method.

The green synthesis of nanoparticle using biological extracts is very rapid and cost effective.

Stachytarpheta jamaicensis is one of the important plants with high medicinal and nutraceutical benefits.

Green Synthesis of Stachytarpheta jamaicensis by using Silver nitrate solution showed highest activity

against E.coli (13mm), S.epidermis (11mm) and fungal strains Candida (12mm). Antimicrobial

activity of Stachytarpheta jamaicensis extract showed less activity such as Pseudomonas (6mm),

Staphylococcus aureus (8mm), Enterococcus (6mm) and fungal species such as Aspergillus (6mm). The

EDAX studies and SEM studies showed the morphological distribution and elemental composition of

Stachytarpheta jamaicensis.

1. Introduction

India has a rich wealth of medicinal plants. Over 7,500 species of plants are estimated to be used by

4,635 ethnic communities for human and veterinary health care across the various ecosystems from

Ladakh in trans Himalayas to the southern coastal tip of kanyakumari and from the desert of Rajasthan

and Karachi to the hills of the North-East India[1]. About 5 to more than 50 million species of plants are

present on the earth [2], accurately of 13.6 million species [3]. Only 1100 of the world’s 365,000 known

species of plants are examined for their medicinal properties [4]. Medicinal plants were harvested from

the wild since ancient times [5]. Among rural communities, where primary health care system are not

provided, traditional medicines(TM) are followed [6].

Sometimes the synthesis of nanoparticles using various plants and their extracts can be

advantageous over biological synthesis process which involves a very complex procedure of maintaining

Compliance Engineering Journal

Volume 10, Issue 12, 2019

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microbial cultures [7].The synthesis of nanoparticles from plant is rapid, low-cost, eco-friendly and a

single method. The green synthesis of nanoparticle using biological extracts is very rapid and cost

effective.Biosynthesis of nanoparticles is a kind of reaction occurring reduction or oxidation. The need

for biosynthesis of nanoparticles is the physical and chemical processes were costly. Often, chemical

synthesis method leads to presence of some toxic chemical substances which adversely affect in medical

applications[8-14]. The pathway for nanoparticle synthesis, scientist used microbial enzymes and plant

extracts (phytochemicals). With their antioxidant or reducing properties they are responsible for the

reduction of metal compounds into their respective nanoparticles[15]. The green chemistry will relation to

bioorganic, inorganic, analytical and even physical chemistry emphasizes only on large scale applications

[16-17].

2. Materials and methods 2.1 Preparation of plant sample

The plant Stachytarpheta jamaicensis was

collected from Lekshmipuram college Campus Neyyoor,

Kanyakumari District, Tamil Nadu, India. The leaves of

the plant used for this study were rinsed severally with

clean tap water and shade dried in a dark place at room

temperature for few days. The dried plant parts were cut

into small pieces ground in electric chopper to get fine

powder for further use.

2.2. Scientific classification Kingdom – Plantae, Figure.1. Stachytarpheta jamaicensis Clade – Angiosperms Clade – Asterids Clade – Eudicots Order – Lamiales Family – Verbenacae Genus – Stachytarpheta Species – S.jamaicensis Binomial name - Stachytarpheta jamaicensis

2.3. Green synthesis of nanoparticles

2.3.1. Preparation of Plant Extract



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The leaves extract was prepared by taking 100g of leaves. The leaves were washed with several

times with deionized water to remove the dust particles and cut it into small pieces. Then the plant extract

was prepared by boiling in deionized water with 350ml in a beaker. After few hours the aqueous extract

was filtered through the Whatmann No.1 filtrate paper and is stored at 40c for further experiments.

2.3.2. Preparation of silver nitrate solution

Analytical grade silver nitrate solution was purchased from Merk (Indian) Ltd.

0.1 M silver nitrate solution was prepared in 100ml standard measuring flask using distilled water. The

solution was diluted as required and all the solutions were kept away from light and kept in dark.

2.3.3. Synthesis of silver nanoparticles using Stachytarpheta jamaicensis Extract:

5ml of Stachytarpheta jamaicensis extract was treated to the 0.1M of 10ml aqueous solution of

silver nitrate and stirring continued for 1 minute at room temperature.

The solution was changed from light yellow to brown which indicates the formation of silver

nanoparticles. The procedure was repeated for 5ml, 10ml, 15ml, and 20ml extract but the metal used only

10ml AgNo3 throughout this experiment. The prepared silver nanoparticles was centrifuged at 15minutes

at 15,000 rpm from that supernatant was discarded and the pellet was collected. The collected pellet was

dried at room temperature for two weeks. The dried biomass was taken for further characterization of

study.

2.4. Antimicrobial activity

2.4.1. Test microorganisms

Five bacterial strains namely Escherichia coli, Enterococcus sp, Klebsiella pneumonia,

Staphylococcus epidermidis and Staphyloccocus aureus and two fungal strains namely Aspergilus niger

and Candida sp were used in this investigation.

2.4.2. Inoculum preparation

The media used for antibacterial test were Nutrient Broth. The test bacterial strains were

inoculated into nutrient broth and incubated at 37oC for 24hrs. After the incubation period, the culture

tubes were compared with the turbidity standard. Fungal inoculums were prepared by suspending the

spores of fungus (as previously cultured) in saline water mixed thoroughly, made turbidity standard and

used.

2.4.3. Assay



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Bioassay was carried out by Agar well diffusion method. Fresh bacterial culture of 0.1 ml having

108 CFU was spread on nutrient agar (NA) plate using swab. The fungal strains also the same but the

medium was Potato dextrose agar (PDA). Wells of 6 mm diameter were punched off into medium with

sterile cork borer and filled with 50µl of plant extracts by using micro pipette in each well in aseptic

condition. Plates were then kept in a refrigerator to allow pre-diffusion of extract for 30 minutes. Further

the plates were incubated in an incubator at 37oC for 24hours and 28-30oC for 3-4 days for bacterial and

fungal cultures respectively. The antimicrobial activity was evaluated by measuring the zone of

inhibition.

2.5. Characterization study

For the characterization study of silver, lead and copper nanoparticles, the following instruments are necessary to confirm the production of nanoparticles. Hence the following instruments are used to characterize silver nanoparticles synthesized by green methods.

i) Scanning Electron Microscopy (SEM)

ii) Energy Dispersive X-Ray Spectroscopy(EDAX)

2.5.1. SEM/EDAX analysis

The surface and morphology of biologically synthesized silver nano particles was studied by

means of SEM. Scanning Electron Microscopy is a technique that uses electrons instead of light to form

an output image. Energy Dispersive X-Ray spectroscopy is a technique that provides the elemental curve

as output. EDX technique primarily detects the X-Rays emitted from the sample during the process of

bombardment by an electron beam for characterizing the elemental composition of the sample of interest.

EDAX helps to verify silver in the sample and the percentage as well. Microstructures were examined by

Scanning Electron Microscope (SEM) with JEOL JSM 6390 model. The chemical composition was

determined by an Energy Dispersive X-ray Spectroscopy (EDAX) attached to SEM.

3. Results and Discussion

3.1. Antimicrobial Property of Stachytarpheta jamaicensis in silver Nanoparticle

Green synthesis of Stachytarpheta jamaicensis plant extract was prepared with Silver nitrate,

solutions. The antimicrobial activity of this metal nanoparticle was studied against bacterial strains such

as E.coli, Pseudomonas, S.epidermis, Staphylococcus aureus, Enterococcus and fungal strain such as

Aspergillus , Candida by well diffusion method. After 2-3 days of incubation at room temperature, the

antimicrobial acivity was evaluated by measuring the zone of inhibition.



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Figure- 2 E.Coli Figure- 3 Pseudomonas Figure -4 S.epidermis

Figure- 5 Staphylococcusaureus Figure- 6 Enterococcus Figure- 7 Aspergillus

Table -1 Antimicrobial Activity of Silver Nitrate

Figure- 8

Candida

In this study, Green Synthesis of Stachytarpheta jamaicensis by using Silver nitrate solution

showed highest activity against E.coli (13mm), S.epidermis (11mm) and fungal strains Candida

SL. No. Test organisms Plant extract with Silver Nitrate

1 E.coli 13 mm 2 Pseudomonas 6 mm 3 S.epidermis 11 mm 4 Staphylococcus aureus 8 mm 5 Enterococcus 6 mm 6 Aspergillus 6 mm

7 Candida 12 mm



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(12mm). Antimicrobial activity of Stachytarpheta jamaicensis extract showed less activity such as

Pseudomonas (6mm), Staphylococcus aureus (8mm), Enterococcus (6mm) and fungal species such as

Aspergillus (6mm) ( Table 1) (Figures 2-8).

3.2. Scanning Electron Microscopy Analysis of Stachytarpheta jamaicensis in silver Nanoparticle

The SEM images give different morphologies like tubular and needle shaped nanocrystals. The particles

with equivalent diameter ranging from o.2µm to 5µm were observed. The fractions of the particle within

the size range of 38.66nm and 64.74nm.

Figure 9. SEM images of Stachytarpheta jamaicensis in silver Nanoparticle

3.3. Energy Dispersive X-Ray Spectroscopy Stachytarpheta jamaicensis in silver Nanoparticle

The EDAX techniques detects X-Ray emitted from the sample during bombardment by an electron beam

to characterize the elemental composition of the material were analyzed with high resolution. EDAX-

analysis data confirms the main component of the material. The weight percentage of silver nanoparticles

was synthesized using Stachytarpheta jamaicensis extract is 54.48%. The strong peak observed in the

spectrum was related to O- 35.38%, Cl - 6.30%, K - 2.19%, Ca - 1.65%. The EDAX spectrum shows the

presence of dominant element of Stachytarpheta jamaicensis sample was represented in the figure 10,

(Table 2).



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FIGURE- 10 EDAX Analysis

Table- 2 EDAX-Analysis of silver Stachytarpheta jamaicensis nanoparticles

4. Conclusion

Stachytarpheta jamaicensis belongs to the family of Verbenaceae, it is one of the important plant

with high medicinal and nutraceutical benefits. In this study Green Synthesis of Stachytarpheta

jamaicensis by using Silver nitrate solution showed highest activity against E.coli, S.epidermis and fungal

strain Candida. SEM images of Silver nanoparticles showed tubular and needle shaped crystals ranges

from 38nm to 64nm. The EDAX technique detects the X-Rays emitted from the sample during

bombardment by an electron beam to characterize the elemental composition of materials was analyses

with high resolution. The weight of the silver nanoparticles is synthesized by using Stachytarpheta

jamaicensis is 54.48%.

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exist. Advances in economic Botany ,vol.12. The New York Botanical Garden,New York.

Element App Conc

Intensity Corrn

Weight% Weight% Sigma

Atomic%

O K 11.82 0.3842 35.38 2.75 73.93

Cl K 5.04 0.9210 6.30 0.38 5.94

K K 2.22 1.1637 2.19 0.30 1.87

Ca K 1.22 0.8473 1.65 0.23 1.38

Ag K 41.82 0.8828 54.48 2.37 16.88

Totals 100.00



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