Biosynthesis of Copper Nanoarticle using Nerium oleander

 

Devika R*, Shalini Chauhan, Subbaiya R

Department of Biotechnology, Aarupadai Veedu Institute of Technology, Vinayaka Missions Research Foundation (Deemed to be University), Paiyanoor – 603104.

 

ABSTRACT:

Nerium oleander leaves were collected from Sipcot IT Park and the leaf extract was prepared withdeionized water and used for the synthesis of copper nanoparticles.  The colour of the solution changes from yellowish green to dark brown, which confirmed the formation of copper nanoparticles. CuNPs were characterized by UV-Vis spectroscopy from which absorbance peak was found at 320-620 nm. CuNPs confirmed natural bio compounds found in the leaf extract by FTIR peaks that are responsible for reducing and stabilizing agent. SEM micrographs of the synthesized by the reduction of copper sulphate revealed spherical and crystalline which ranged in an average size of 20 nm. Biological method found to be eco-friendly, cost effective method, which provided natural capping agent.

 

 

INTRODUCTION:

Nanotechnology is the science and technology in which the structure of matter is controlled at the nanometer scale, this field is making an impact on human life such as pharmaceutical, health, food, electronics, chemical industry, energy science, cosmetics, and environmental science and space industries etc.¹ Nanotechnology is an important field of modern research dealing with synthesis, strategy and manipulation of particles structure ranging from approximately 1 to 100 nm in size. Eco-friendly processes in biological and chemical technologies are becoming increasingly popular and are needed to solve problems associated with environmental concerns². A nanomaterial major role in field of high sensitivity bio molecular detection, catalysis biosensors, medicine, cosmetics, food and feed, drug delivery, space industries, light emitters and photo-chemical applications and also exhibits a strong inhibitory and bactericidal effect, anti-fungal, anti-inflammatory and anti- angiogenesis activities³.

 

 

Nanoparticles can be synthesized using various approaches including chemical, physical and biological, for synthesis of large quantity of Nanoparticles⁴. Biological method is one of the best methods for the synthesis of Nanoparticles. This method has several advantages namely low cost, simple use of less toxic materials and eco-friendly⁵.

 

Plants are known to possess various therapeutic compounds which are being exploited since ancient time as a traditional medicine⁶. Its huge diversity plants have been explored constantly for wide range of applications in the field of pharmaceutical, agricultural, industrial etc.⁷and reported that plants towards the production of nanoparticles because of easy availability, safe to handle and broad range of biomolecules such as alkaloids, terpenoids, phenols, flavanoids, tannins, quinones etc.⁸. Nano scale materials such as nanoadsorbents, nanocatalysts, nanofiltration, and nanobiocides such as metal and metal oxide nanoparticles are currently being employed for remediation of water and wastewater pollutants⁹. Silver nanoparticles were synthesized from extract of dried leaves of Catharanthus roseus  which showed anti cancer effect on cancer cell lines¹⁰.

 

 

Nerium oleander is an evergreen shrub in the dogbane family Apocynaceae and it is the only species currently classified in the genus Nerium¹¹. The leaves and the flowers are reported to be cardiotonic, diaphoretic¹², diuretic, emetic, expectorant and sternutatory effects in the medicinal field. The root is known as powerful resolvent¹³ because of its poisonous nature it is only used externally¹⁴.  

 

MATERIALS AND METHOD:

Collection of plant leaf:

Nerium oleander leaf was collected from Sipcot I.T Park. The collected leaves were tightly packed with polythene bag and then transferred to the laboratory. Then it was washed several times with distilled water.The plant leaf extract was prepared by using Nerium oleander fresh leaves (5g) and dissolved in 100ml of distilled water and made into homogenizedsolution by continuous magneticstirring for 1 hr¹⁵.  The extract was filtered by Whatmann No.1 filter paper.

 

Synthesis of copper nanoparticles:

During synthesis of copper Nanoparticles, 1Mm CuSo₄ solution and leaf extract were taken. To reduce the Cu++ ions, 20 ml of leaf extract was added drop wise to 10 ml of 1Mm CuSo4 solution, with continuous stirring using magnetic stirrer. The color change was observed from very brown color to green after 24 hrs at room temperature.

 

The solution of centrifuged and pellet was taken, the pellet was washed with water and centrifuged again and the resultant pellet was taken and dried in the oven. The sample was stored in a sterile air tight glass bottle and it was then used for the FTIR and SEM analysis.

 

CHARACTERIZATION STUDIES:

UV-spectra analysis:

Sample (1ml) of the suspension was collected periodically to monitor the completion of bio-reduction of Cu++ ions in aqueous solution followed by dilution water and subsequent UV measurements. The absorbance was measured on a TRIAD spectrophotometer between 320-20 nm¹⁶.

 

FTIR:

To determine the bio-molecules present in the leaf extract. FTIR analysis was carried out which is responsible for the reduction of Cu ions with the spectral range 500 to 4000cm-1 [10] , to confirm the formation of the ether linkage between the molecules¹⁷.

 

SEM:

The sample was prepared by drop casting 5µl of the sample on a carbon coated copper grids and air drying. SEM analysis was used to study, the surface morphology modification of nanoparticles formed.

 

RESULTS AND DISCUSSION:

The plant leaf extract was prepared by using Nerium oleander fresh leaves (5g) and dissolved in 100ml of distilled water and made into homogenized solution by continuous magnetic stirring for 1 hr¹⁵.  The extract was filtered by Whatman No.1 filter paper. Biosynthesis of Cu nanoparticles occurred by the reduction of Cu++ ions in 20 ml of leaf extract was added drop wise to 10 ml of 1Mm CuSo4 solution, with continuous stirring using magnetic stirrer. The color change was observed from very brown color to green after 24 hrs at room temperature Fig. 1. Silver nanoparticles were formed rapidly with 30 minutes using leaf extract of Acalypha indica¹⁸. A stable and spherical shaped silver nanoparticles were developed using Abutilon indicum which had high antimicrobial effect on various species¹⁹. Very recently biosynthesis of nanoparticles are common from various plants such as Solanum tricobatum²⁰, Cochlospermum religiosum²¹, Erythrina indica²² and the size of nanoparticles ranged from 5-20nm.

 

 

Fig 1: Synthesis of CU nanoparticles from Neriumoleander

Sample (1ml) of the suspension was collected periodically to monitor the completion of bio-reduction of Cu++ ions in aqueous solution followed by dilution water and subsequent UV measurements. The absorbance was measured on a TRIAD spectrophotometer between 320-20 nm¹⁶ and the result is represented in Fig.2. The formation of CuNPs was confirmed by UV-Vis spectra analysis and a broad absorption peak was observed between 320-620 nm. The size distribution histogram of silver nanoparticles synthesized from Azadirachta indica leaf extract was reported to be 34 nm²³.

 

 

Fig 2: UV- Vis spectroscopy analysis of CU nanoparticles from Neriumoleander

 

The FTIR spectrum of plant (Nerium sps.) mediated 1Cu Nanoparticles showed that the broad absorption band at 3698.28cm-1 corresponded to O-H functional groups, 2926.33 cm-1 had C-H functional groups Fig.3. The IRspectrum of CuNPs showed band at 3473 cm-1, 16535cm-1, 516 cm-1,1376cm-1, 1178 cm-1, corresponded to O-H stretching H-bonded alcohols, phenols, carbonyl stretching N-H bond, primary amines , corresponds to C-N  stretching of the aromatic amine group and C-O stretching alcohols, ethers, respectively²⁴. FTIR spectrum of Cu Nanoparticles was surrounded by different organic molecules such as terpenoid, alcohols, ketones, aldehydes, and carboxylic acid. Silver nanoparticles from Ziziphor atemniar leaves were characteristised under Tem and it showed uniform spherical structures ranging from 8-40nm with carbonyl, hydroxyl and other stabiliising functional groups under FTIR analysis²⁵. FTIR results of silver nanoparticles from Azadirachta indica showed a broad band between 3454 cm^-1 due to N-H stretching vibration of group NH₂ and OH overlapping of the stretching vibration of water and leaf extract molecule of A. indica. Further,  peaks were observed at1113 cm-1 denoted by –c-oc- linkages or –c-o bond which was attributed to flavonoids and terpenoids which were high in the leaf of A. indica²⁶.

 

 

Fig 3: FTIR analysis result of CU nanoparticles from Neriumoleander

 

SEM analysis of nanoparticle is used to identify the size, shape and morphology. It was reported that the surface morphology of the CuNPsin the present study under SEM micrographs the CuNPs synthesized revealed uniform spherical shape Fig. 4. It was reported that the silver nanoparticles from A. indica showed predominantly spherical in shape and seems to be homogenous²⁶.Stable silver nanoparticles at different concentration gave a distinct spherical shaped particles ranging from 15 to 50 nm from Acalypha indica. TEM image of green silver nanoparticles from Citrus sinensis showed very well dispersed spherical particles of 3-12 nm size with a highest fraction diameter of 6nm²⁷.

 

 

Fig 4: SEM analysis result of CU nanoparticles from Neriumoleander

CONCLUSION:

From the present investigation, green and biosynthesis of Cu Nanoparticles using leaf extract of Nerium oleandershowed the formation of significant nanoparticles characterization through various instrumental analysis. Nanoparticles are proved to be more advantageous over other reported traditional methods which are having more disadvantages as expensive, process is not that simple, use of toxic reagent etc. Further on investigation of these nanoparticles may pave a way for future use in the medical field for curing many diseases.

 

REFRENCES:

1.     Singh P, Katyal A, Katra R, Chandra R. Biological synthesis of triangular nanoparticles. Tetrahedron Letter. 49; 2008: 727-725.

2.     Thuesombat P, Hannonngbua S, Akasit S and Chad chawan S. Ecotoxicology and environmental safety effect of silver nanoparticles on rice (Oryza sativa L) seed germination and seedling growth. Exotoxicology and Environmental Safety. 1043; 2014: 302- 309.

3.     Chowdary S, Ghosh AK, Maji S and Bala NN. Antioxidant capacities of a flavonoid compound isolated from C. salignus leaves. International Journal of Pharma and Biosciences. 3(4); 2014: 112- 118.

4.     Jana NR, Wang ZL, Sau TK, Pal T. Seed mediated growth method to prepare cubic copper Nanoparticles. Current Science. 79; 2003: 1367-1370.

5.     Bharde,  Parikh RY, Bai.dakova, Joven M, Hannoyer S, Enoki B, Prasad T, Shouche B, Sastry S and Langmuir M.  Role of nanotechnology in novel drug delivery system. Journal of Pharmaceutical Science and Technology. 24; 2008: 5787-5794.

6.     Lisichkin GV, Russ VK, Sharma RA Yingard, Y Lin.  Extracellular biosynthesis and antimicrobial activity of silver nanoparticles. Advance Colloidal International Science. 45; 2009: 83-86.

7.     Gnanajobitha G, Annadurai G and Kannan C. Green synthesis of silver nanoparticles using Elettaria cardamomom and assessment of its antimicrobial activity. International Journal of  Pharma Science Research. 3; 2012: 330-323.

8.     Dinesh S, Karthikeyan S and Arumugam P. Biosynthesis of silver nanoparticles from Glycyrrhiza glabra root extract. Arch Applied Science Research, 4; 2012: 178-187.

9.     Chethan J, Sampath Kumar KK, Shailashree Sekhar and Prakash HS, Antioxidant, antibacterial and DNA protecting activity of selected medicinally important Asteraceae plants. International Journal of Pharmacy and Pharmaceutical Sciences. 4(2); 2012: 975- 981.

10.   Ghozail M, Patil Jai. Biosynthesis of silver nanoparticles and their effect on anti-cancerous cell –lines. International Journal of Nanoscience. 5; 2015: 12-28. 

11.   Henary HA, Kurzrock R Falchook, GS, Naing A, Moulder SL, Wheler JJ, Tsimberidou AM, Durand J, Yang P, Johansen MJ, Newman R, Khan R, Patel U and Hong DS. Final results of a first-in-human phase I trial of PBI-05204, an inhibitor of AKT, FGF-2, NF-Kb, and p70S6K in advanced cancer patients. Journal of Clinical Oncology. 29 (15); 2011: 3023-3025. 

12.   Newman RA, Yang P, Pawlus AD and Block KI. Cardiac Glycosides as Novel Cancer Therapeutic Agents.  Molecular Interventions. 8 (1); 2008: 36–49.

13.   Szabuniewicz M, McCrady JD and Camp BJ. Treatment of experimentally induced oleander poisoning.  Archives Internationales de Pharmacodynamie Etde Therapie. 189(1); 2001: 12–21.

14.   Shankar S, Rai A, Ahmad A and Sastry M. Rapid synthesis of Au, Ag and bimetallic Au core Ag shell nanoparticles using Neem leaf broth. Journal of colloidal Interference Sciences. 275; 2004: 496-502.

15.   Gopinath M, Subbaiya R, Masilamani M, Selvam and suresh D. Synthesis of copper Nanoparticles from Nerium oleander leaf aqueous leaf extract and its antibacterial activity. International Journal of Current Microbial Applications and Sciences. 3(9); 2014: 814-818.

16.   Lalitha A, Subbaiya R and Ponmurugan. Green synthesis of silver Nanoparticles from leaf extract Azhadachta indca. International Journal of Current Microbial Applications and Sciences.2; 2013: 228-235.

17.   TakayamaS, LinkG and SatoM. Microwave and radio frequency applications in proceeding of the fourth world congress on microwave and radio frequency application., 3; 2004: 311-318.

18.   Krishnaraj C, Japan EG and Rajasekar S. Synthesis of silver nanoparticles using Aclyphaindica leaf extracts and its antibacterial activity against water borne pathogens. Colloids Surf B. 76; 2010: 50-56.

19.   Ashok Kumar, Ravi S, Kathirvan V and Velmurugan S. Synthesis of silver nanoparticles usins A. idicum leaf extract and their antibacterial activity. Spectrochem Acta Part A; Molecular Spectroscopy. 134; 2015: 34-39.

20.   Logeshwari P, Silambarasan S and Abraham J. Synthesis of silver nanoparticles using plant extract and analysis of their antibacterial property. Journal of Saudi Chemical Society. 19(3); 2012: 312-317.

21.   Sasikala A, Linga Rao M, Savithramma N and Prasad TNVKV. Synthesis of silver nanoparticles from stem bark of Cachlospermum religiosum(L) Aston: an important medicinal plant and evaluation of their antimicrobial efficacy. Applied Nanoscience. 10; 2014: 1007-1010.

22.   SrePRR, Reka M, Poovazhagi R, Kumar MA and Murugesan. Antibacterial and cytotoxic effect of biologically synthesized silver nanoparticles using aqueous root extract of Erythrina indica. biomolecular spectroscopy. 135; 2015: 1137-1144.

23.   Shakeel Ahmed, Saifullah, Medasir Ahmed, Babu Lal Swami and Saqa Ikram. Green synthesis of silver nanoparticles using Azadirachta indica aqueous leaf extract. Journal of Radiation Research and Applied Science. 9; 2015: 1-7.

24.   RaveendraP and walker J. Role of biopolymers in green nanotechnology. Journal of American Chemical Sciences. 125; 2013: 13940-13941.

25.   Awwad AM, Salem NM and Abdeen AO. Biosynthesis of silver Nano using olea europaea leaf extract and its antibacterial activity Guobiene A- Analysis of silver Nanoparticles produced, by chemical reduction of silver salt solution. Material Science.  2(4); 2006: 1916-1920.

26.   Sadeghi B, Cholamhoseinpoor F. A study on the stability and green synthesis of silver nanoparticles using Ziziphora termour at room temperature. Spectrochem Acta Part A; Molecular Spectroscopy. 134; 2015: 310-315.

27.   Ramyal M, Subapriya MS. Green synthesis of silver nanoparticles. International Journal Pharma Medical Biological Sciences. 1(1); 2012: 54-61.

 

 

 

 

 

Accepted on 18.12.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(2):621-624.