Senna Auriculata Flower Extract mediated green synthesis of iron nanoparticles and their Antibacterial activity

K. Sowmiya, J. Thomas Joseph Prakash*, D. Devi Priya

PG And Research Department of Physics, Government Arts College

(Affiliated to Bharathidasan university) Tiruchirappalli-620 022, Tamil Nadu, India.

ABSTRACT

Green synthesis of metallic nanoparticles has accumulated an ultimate interest over the last decade due to their distinctive properties that make them applicable in various fields of science and technology. Metal nanoparticles that are synthesized by using plants have emerged as non toxic and ecofriendly. In this study, a very cheap and simple conventional heating method was used to obtain the iron nano particles (Fecl3) using the extract from the flower of senna auriculata plant. The obtained iron nanoparticles were characterized by UV-Vis spectroscopy, FTIR spectroscopy, PXRD, DLS, FESEM, EDAX and the antibacterial activity was studied against staphylococcus aureus, Bacillus Subtilis and Escherichia coli by the standard disc diffusion method.

Key word: Senna Auriculata, Iron nanoparticles, Antibacterial Activity.

* Corresponding Author

J. Thomas Joseph Prakash

PG And Research Department of Physics, Government Arts College

(Affiliated to Bharathidasan university) Tiruchirappalli-620 022, Tamil Nadu, India.

E-Mail: armyjpr1@yahoo.co.in

Cell: 09842470521

1. INTRODUCTION

In recent years, novel quantitative physicochemical characteristics of the most powerful metallic iron nanoparticles applications in a wide range, in the magnetic storage media [1], ferrofluids [2] biosensors [3], the catalyst [4], separating the leading processes, and Environmental Change [ 5]. In particular, magnetite (FeCl3) and having a cubic spinel structure upside down in a common magnetic iron oxide. The composition of the octahedral sites [6] on the basis of the transfer of electrons between Fe2+ and Fe3+ reveals the unique electrical and magnetic properties.

Their unique physical, chemical, thermal and mechanical properties, hyperbarermia, and magnetic flow, such as the superior parallel applications, such as super paragomic nanoparticles for cellular therapy, tissue repair, pharmaceutical supplies, and magnetic resonance imaging (MRI) 7]. Currently, physics, chemistry major, biology, and hybrid methods available to synthesize various types of nanoparticles [9]. Specific characteristics of nanoparticles are formed using each method. However the plants biosynthesis of the metal nanoparticles is currently under development. Green Nano technology has attracted attention and has widespread practices that reduce the environment or eliminate the remaining toxins. A modern alternative to the production of plant nutrients, 10 of the nano particles, [11] in tapacar sauces, high fatty acid [12] living plants and other areas [13] using plant tissue. Nano particles of green node of environmental friendly non-toxic and safe chocolate [14] applications.

2. MATERIAL AND METHODS

A. Materials

Iron (III) chloride/ Ferric chloride (FeCl3) analytical grade was purchased from Merck & Co. and used without further purification. Double distilled de-ionised (DI) water was used throughout the course of this investigation. A solution of FeCl3 (0.01 M) was prepared by dissolving the solid FeCl3 in DI water. The senna auriculata flowers (Fig 1) were freshly collected from Trichy district, Tamilnadu, India

B. Synthesis of Iron nanoparticles

The fresh flowers of senna auriculata broth solution were prepared by taking 100g of thoroughly washed and leaf extract were dried in shade to remove the moisture content and powder in a blender . Finely 5g senna auriculata dried flower in a Erlenmeyer flask along with 500ml of sterilized double distilled water and then boiling the mixture for 15min before finally decanting it. The extract was filtered through Whatmann filter paper no 1 and stored at -4°C. The filtrate was treated with an aqueous 1mM FeCl3 solution in an Erlenmeyer flask and the mixture was heated at 60°-70°C for 10-15 minutes. As a result, a black colored solution was formed; indicating the formation of Iron nanoparticles. The Iron nanoparticle solution thus obtained was purified by repeated centrifugation at 10,000 rpm for 10 min. As a black color solution was further confirmed by UV-Visible spectrum analysis. It showed that aqueous Iron ions could be reduced by aqueous extract of plant parts to generate extremely stable Iron nanoparticles in water (Fig 2).

3. MEASUREMENTS

A. UV-Vis spectra Analysis

Samples (1 mL) of the suspension were collected periodically to monitor the completion of bio-reduction of Fe3+ in aqueous solution, followed by dilution of the samples with 2 ml of deionized water and subsequent scan in UV-visible (vis) spectra, between wavelengths of 200 to 700 nm in a spectrophotometer ( Perkin – Elmer Lambda ), having a resolution of 1 nm.

B. Fourier Transmission Infra Red spectroscopy ( FTIR)

Fourier transform infrared (FT-IR) spectroscopy was done to categorize the functional groups present in synthesized iron oxide nanoparticles.

C. Dynamic light scattering (DLS)

Partical size analysis was carried out using the dynamic light scattering ( NANO PLUS ) in polysterene cuvette

D. Powder X-Ray difractometry study ( P-XRD)

The synthesized iron oxide nanoparticles were dried at 50◦ C to carry out the XRD studies. The diffraction pattern was recorded by X PETRO PRO – X-ray diffractometer. which provides control modules for the complete range of difractometer accessories together with the corresponding analysis software.

E .FESEM and EDX Analysis

The samples have been analyzed by the Quanta-200F SEM. The dual side carbon tape acetone was fixed on the clean glossy aluminum (10 mm - 10 mm - 5mm) piece. The samples were spread among the carbon tapes and crushed with the pressure and free particles to remove with the butter sheet. The models were placed in a vacuum room for 1 hour and placed on the SEM tool. Inorganic metals with samples identified with EDS classification

F. Antibacterial activity of samples (disc diffusion method)

Antibacterial activity of samples was determined using the disc diffusion method. The petridishes (diameter 60 mm) was prepared with Muller Hinton Agar and inoculated with test organisms. Sterile disc of six millimeter width were impregnated with 10 µl of various samples respectively. Prepared discs were placed onto the top layer of the agar plates and left for 30 minute at room temperature for compound diffusion. Positive control was prepared using the 10 µl of Amoxicillin as standard antibiotic disc. The dishes were incubated for 24 h at 37ºC and the zone of inhibition was recorded in millimeters and the experiment was repeated twice.

4.RESULT AND DISCUSSION

A.UV- Visible Analysis

The most widely used technique to investigate the optical properties of UV-visible spectroscopy particles. The color change from brown to black indicates the formation of iron nano particles. UV - paralysis spectroscopy analysis was done in 200-400nm [15,16] range and maximum absorption was found in the production of iron nano particles due to the enthusiasm of surface plasma vibrations in the 268nm (Fig 3)

B.FTIR Analysis

FTIR is analyzed to know the biomolecules responsible for causing of Fe3+ ions in the ferric chloride and the shelter of ferric nanoparticles compiled by the source flower juice extract. The effect of FTIR shows a intensive peak located at 3413 cm-1, 2934 cm-1, 1632 cm-1, 1400 cm-1, 1109 cm-1, 867 cm-1, 800 cm-1, 764 cm -1 and 619 cm-1 as shown in (Table 1) [17]. It is related to the functional groups in the set and elements of the ferric nano particles. As shown in (Fig. 4).

C . Dynamic light scattering (DLS)

The models revealed that DLS analyzes and hydrodynamic diameter is 85nm range (Fig 5)

D.PXRD Analysis

The phase identification and crystalline structures of the nanoparticles was characterized by X-ray powder diffraction. The X-ray diffraction patterns obtained for the FeCl3-NPs synthesized using senna auriculata extract is shown in (Fig 6). It is found that there exist strong diffraction peaks with 2θ values of 30.4°, 35.8°, 43.5°, 54.1° and 57.4°, corresponding to the crystal planes of (200), (311), (511) and (440) of crystalline FeCl3-NPs, respectively [18,19]. The results show the spinel phase structure of magnetite and are in agreement with the XRD standard for the magnetite nanoparticles .

E.FESEM and EDX Analysis

The prepared iron nanoparticles were analyzed by scanning Field Electron microscopy (FESEM) with energy dispersive X-ray spectroscopy (EDX) to know the morphology and atomic percentage (fig 7) shows the FESEM image 100 ml of 0.01M aqueous Fecl3 solution with 20 ml of senna auriculate leaf extract. FESEM image shows the clear morphology of nanoparticles. The FESEM also reveals that average size of nanoparticles were in the range of 30nm – 100nm . Energy – dispersive X-ray spectroscopy is an analytical technique used for the elemental analysis. The EDX spectrum (fig 8) contains intense peaks of cl and C in addition to Fe and O. The c signals are attributed mainly to organic molecules in the senna auriculata leaf extracts. The atomic percentages obtained from EDX quantification were 48.98% of O, 8.83% of Fe, 36.77% of C and 5.42% of cl. The values could be helpful in reflecting the atomic content on the surface and near surface regions of the NPS .

F.Antibacterial Activity

Disc Diffusion Assay

The results of the antibacterial activity of various samples were tested against pathogens by disk diffusion method are shown in (Table 2). The Sample D showed growth inhibitory activity against Staphylococcus aureus (6 mm). At sample C exhibited the antibacterial activity all the four bacteria, but was more susceptible against Escherichia coli (5 mm), Bacillus subtilis (4 mm). However, the crude extract and synthesized nanoparticles showed better inhibitory actions against pathogens (Fig 9).

5. CONCLUSION

Senna auriculata provides an eco-friendly, simple and efficient way of quick biological synthesis of iron nano particles using flower. Under UV-visible wavelength, nanoparticles showed a surface plasma resonance behavior. Color change was remarkable when mixing with agent for reducing the mixture of ferric chloride. The Biosynthesized FeNPs were chareacterized by the UV-Vis spectroscopy, FTIR spectroscopy, DLS, PXRD, FESEM-EDAX and anti-bacterial activity. Overall, this approach assures most of the green stability of FeNPs.

REFERENCES

[1] Pattanayak, Monalisa, P. L. Nayak, “ Ecofriendly Green Synthesis of Iron Nanoparticles from various Plant and Spices Extract” International Journal of Plant, Animal and Environmental Sciences, 2013, 3(1), 68-78.

[2] M. Herlekar, S. Barve, R. Kumar,’’Plant – Mediated Green Synthesis of Iron NanoParticles” Journal of Nanoparticles, 2014, 2014, 1-9.

[3] Mahdavi, Mahnaz, et al “Green biosynthesis and characterization of magnetic iron oxide (Fe3O4) nanoparticles using seaweed (Sargassum muticum) aqueous extract”, Molecules. 2013, 18(5), 5954-5964.

[4] S. Machado, S.L. Pinto, J. P. Grosso, H. P. A. Nouws, J. T. Albergaria, C. Delerue-Matos, “ Green production of zero – valent Iron nanoparticles using tree leaf extracts ”,Science of the total environment, 2013, 445 - 446, 1-8.

[5] Monalisa Pattanayak, P. L. Nayak, “ Green Synthesis and characterization of zero valent Iron Nanoparticles from the leaf extract of azadivachta indica (Neem) ” World Journal of Nano Science & Technology, 2013, 2(1), 06-09.

[6] T. Wang, J. Lin, Z. Chen, M. Megharaj, R. Naidu, “ Green synthesized iron nanoparticles by green tea and eucalyptus leaves extracts used for removal of nitrate in aqueous solution ” Journal of Cleaner Production. 2014, 83, 413-419.

[7] Y. Liu, S.A. Majetich, R. D. Tilton, D. S. Sholl, G. V. Lowry, “ TCE Dechlorination Rates, Pathways, and efficiency of Nanoscale Iron particles with Different properties ” Environmental Science & Technology. 2005, 39 (5), 1338–1345.

[8] Awwad, Akl M., & Nidá M. Salem.” A green and facile approach for synthesis of magnetite nanoparticles”, Nanoscience Nanotechnology. 2012, 2(6), 208-213.

[9] S. Machado, JG. Pacheco, HP. Nouws, JT. Albergaria, C. Delerue-Matos, “ Characterization of Green zero – valent iron Nanoparticles produced with tree leaf extracts ” Science of the total environment. 2015, 15(533), 76-81.

[10]. Salam, H.A.; Rajiv, P.; Kamaraj, M.; Jagadeeswaran, P.; Gunalan, S.; Sivaraj, R , ”Plants: Green route for nanoparticle synthesis”, International Journal of Biological Sciences, 2012, 1, 85–90

[11] Raveendran, Poovathinthodiyil, Jie Fu, & Scott L. Wallen.” Completely green synthesis and stabilization of metal nanoparticles”, Journal of American Chemical Society, 2003, 125(46), 13940-13941.

[12] A. K. Tyagi and A. Malik, “ Antimicrobial potential and chemical composition of Eucalyptus globulus oil in liquid and vapour phase against food spoilage microorganisms ” journal of Food Chemistry. 126, 228 - 235 (2011).

[13] N. Toshima and T. Yonezawa, “ Bimetallic nanoparticles – novel materials for chemical and physical applications ”,New Journal of chemistry. 22, 1179 - 1201 (1998).

[14] Philip, Daizy. “Green synthesis of gold and silver nanoparticles using Hibiscus rosa sinensis”, Physica E: Low-dimensional Systems and Nanostructures, 2010, 42(5), 1417-1424.

[15] Kumar, Amit, et al” Phytochemical, ethnobotanical and pharmacological profile of Lagenaria siceraria:”, Journal of Pharmacognosy and Phytochemistry., 2012, 1, 27-35.

[16] C. W. Lee, S. S. Jung, and J. S. Lee, “ Phase transformation of ᵦ - Fe2O3 hollow nanoparticles ” Material. Letter. 62, 561 (2008).

[17] W. Wu, Q. He, and C. Jiang, “ Magnetic Iron oxide nanoparticles : Synthesis and surface Functionalization strategies ” Nanoscale Research Letters. 3, 397 (2008).

[18] A. Kumar and A. Singhal, “ Synthesis of colloidal ᵝ - Fe2O3 Nanostructures – influence of addition of CO2+ on their morphology and magnetic behavior ” Nanotechnology 18, 475703 (2007).

[19] J. Lee, T. Isobe, and M. Senna, “ Preparation of ultrafine Fe3O4 particles by precipitation in the presence of PVA at high PH ” Journal of Colloid Interface Science. 177, 490 (1996).

FIGURE CAPTIONS

Fig.1 Senna auriculata flower

Fig.2 Colour changed at brown to black

Fig.3 UV-Vis spectrum of FeCl3 -NPs Using senna auriculata flower extract

Fig.4 FT-IR spectrum of FeCl3-NPs Using senna auriculata flower extract

Fig.5 Dynamic light scattering (DLS) of FeCl3-NPs Using senna auriculata flower extract

Fig.6 PXRD pattern of FeCl3– NPs synthesized using supernatant of senna auriculata flower

Fig.7 The typical SEM image of the FeCl3– NPs of senna auriculata flower extract

Fig.8 The EDAX of FeCl3– NPs of senna auriculata flower extract

Fig.9 Anti Bacterial Activity disc diffuce method

TABLE CAPTION

Table .1 FTIR Analysis

Table.2 Antibacterial activity of Tested compounds against Bacterial pathogens

Fig.1 Senna auriculata flower

Fig. 2 Colour change of flower extract after the addition of FeCl3

Fig.3 UV-Vis spectrum of FeCl3 - NPs Using Senna auriculata flower

extract

Fig.4 FT-IR spectrum of FeCl3 - NPs Using Senna auriculata flower extract

Fig.5 Dynamic light scattering (DLS)

Fig.6 PXRD pattern of FeCl3-NPs synthesized using supernatant of Senna auriculata flower extract

Fig.7 The typical SEM images of the iron nanoparticle of Senna auriculata flower extract

Fig.8 The EDAX of iron nanoparticles of Senna auriculata flower extract

Fig.9 Anti Bacterial Activity disc diffuce method

Wavenumber peaks ( cm-1)

Type of vibration

Functional group

3413

Stretching

O-H

2934

Stretching

C-H

1632

Bending

C=O

1400

Bending

Amine (C-N)

1109

Stretching

Acid (OH)

867

Bending

C-Cl

800

Stretching

-OH

764

Bending

Alkene (C-C)

619

Stretching

Ester ( C-O)

Table 1 : FTIR Analysis

Samples

Concentrations

(µl/ml)

Organisms/Zone of inhibition (mm)

Staphylococcus aureus

Bacillus subtilis

Escherichia

Coli

A (Amoxicillin)

B (Ferric chloride)

C (Plant extract)

D (Nanoparticles)

10

10

10

10

8

0

3

6

8

0

4

5

8

0

5

4

Table.2 Antibacterial activity of Tested compounds against Bacterial pathogens

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