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Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103.
www.ijprsonline.com
ISSN: 2348 –0882
==============================================================================

Biosynthesis Of Silver Nanoparticles Using Curcuma Longa And Their
Antibacterial Activity
S Elumalai1 and R Devika2
1

Associate professor, Dept. of Plant Biology & Plant Biotechnology, Presidency College, Chennai, India.
2
Research scholar, Dept. of Plant Biology & Plant Biotechnology, Presidency College, Chennai, India.

Email: devinanotech25@gmail.com

Abstract
A rapid advance of nanotechnology has the
potential approach for significant improvements in
disease prevention, diagnosis and treatment. In this
article, we report a simple and eco-friendly
biosynthesis of silver nanoparticles (Ag-NPs) using
silver nitrate as metal precursor in Curcuma longa.
These Ag-NPs were characterized by UV–vis
spectroscopy,
and
Transmission
electron
microscopy (TEM). These nanoparticles exhibited
maximum absorbance in specific nano meter range
in UV–vis spectroscopy. TEM micrographs
revealed the formation of well-dispersed Ag-NPs
with its size and morphology. Microbiology assay
founds that Ag-NPs are effective against V.cholera
bacteria. These developments raise exciting
opportunities to diagnose and treat pathogenic mode
of infection based on the various profiles to target
diseases.
Keywords: Silver nanoparticles, C. longa ,
Spectroscopic studies, Antibacterial Activity
1. Introduction
A novel biosynthesis of nanoparticles is an exciting
methods that have attracted significant attention due
to their potential use in many applications, such as
catalysis, drug delivery biosensor [1] antimicrobials
and therapeutics [2,3]. New application of
nanoparticles and nanomaterials are emerging
rapidly [4]. Biological methods of nanoparticles

synthesis using microorganism, enzyme, andplant
or plant extract have been suggested aspossible
ecofriendly alternatives to chemical andphysical
methods [5]. As part of our work, we have observed
that aqueous silver ions, when exposed to the
rhizome extract of C. longa, are reduced in solution,
thereby leading to the formation of an extremely
stable silver particle.
2. Materials and Methods
2.1 Collection and Extract Preparation
The rhizomes of C. longa were rinsed with fresh
seawater and distilled water to remove associated
debris. The cleaned material was then air dried to
dryness in the shade at 30°C. The dried samples
were finely powdered and stored at -20°C until use.
Approximately 10 g of C. longa biomass was taken
in a conical flask containing 100 mL of distilled
water, kept for 24 hrs and then the aqueous solution
components were separated by filtration. To this
solution, AgNO3 (10-3 M) was added and kept for
several hours at 24 hrs Periodically, aliquots of the
reaction solution were removed and the absorptions
were
measured
in
a
Elico
UV-Vis
spectrophotometer.
2.2 Synthesis and Characterization
For the synthesis of Ag- NPs 1ml of rhizome
extracts as test solution were incubated at room
temperature for 1-2 hours. The silver nanoparticle
solution thus obtained was purified by repeated
98
Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103.
www.ijprsonline.com
ISSN: 2348 –0882
==============================================================================

centrifugation at 15,000 rpm for 20min. Supernatant
is discarded and the pellet is dissolved in deionised
water. It is well known that Ag-NPs exhibit
yellowish brown color in aqueous solution due to
excitation of surface plasmon vibrations in Ag-NPs
[6]. The silver nanoparticles were confirmed by
colour changes and qualitatively characterized by
UV-visible spectrophotometer on a Elico UV- Vis
spectrophotometer. The bioreduction of Ag+ ions in
solution
was
monitored
using
UV-Vis
spectroscopyfrom zerotime reading was noted and
double distilled water as blank, and incubated at
culture condition. The samples were withdrawn at
various time intervals and the absorbance was
measured [7].
2.3 UV-Vis spectroscopy analysis
UV-Vis Spectral analysis was done by using
HITACHI U-2900 Spetrophotometer. The UV-Vis
Spectrophotometer analysis reveals the formation of
silver nanoparticles by showing surface Plasmon
resonance at 422 nm. UV-Vis Spectroscopy is one
of the most widely used techniques for structural
characterization of silver nanoparticles. The
absorption spectrum of the brown silver colloids
prepared by hydrazine reduction showed a surface
Plasmon absorption band with a maximum of 422
nm indicating the presence of spherical or roughly
spherical silver nanoparticles [8].
2.4 TEManalysis of silver nanoparticles
Sample for TEM analysis was prepared as
mentioned in IR sample preparations. The sample
was first sonicated (Vibronics VS 80) for 5 min.
Silver nanoparticles was loaded on carbon-coated
copper grids and solvent was allowed to evaporate
under Infra light for 30 min. TEM measurements
were performed on Phillips model CM 20

instrument operated at an accelerating voltage at
200 Kv.
2.5 Antibacterial activity:
Bactericidal effects of Ag-NPs were studied against
Vibrio cholera bacteria. Antimicrobial activity was
demonstrated by modified method described [9].
Then 0.1 ml of the diluted microbial cultures was
spread on sterile nutrient agar plate. The soaked and
dried discs of 6 mm diameter of Whatman filter
paper No: 1 were then placed on the seeded plates
and gently pressed down to ensure contact [10].
Four replicates were placed for control, Ag-NPs,
antibiotics and Ag-NPs combined with antibiotics
Chloramphenicol, in each disc to confirm the
inhibition a zone, and the plates were incubated at
37°C for 24 hours. After incubation period, the
inhibition zone around the discs were measured and
recorded, as the difference in diameter between the
disc (6 mm) and growth free zone of V. cholera
were measured.
2.6 Results & Discussion
Silver nanoparticles were synthesized from AgNO3
solution containing Ag+ ions by treating with the
rhizome extracts. The color of the solution changed
to deep brownish color within 30 min of reaction
with the Ag+ ions. The appearance of the deep
brownish color indicated formation of silver
nanoparticles. The formation of silver nanoparticles
was confirmed by color changes followed by UVVis spectrophotometer analysis. It is generally
recognized that UV-Vis spectroscopy could be used
to examine size and shape-controlled nanoparticles
in aqueous suspensions [11]. The UV-Vis
spectrophotometer proved to be very useful
technique for the analysis of some metal
99
Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103.
www.ijprsonline.com
ISSN: 2348 –0882
==============================================================================

nanoparticles. The UV- visible spectra (shown in
Fig 1) indicated a strong Plasmon resonance that
was located at ~422 nm. Presence of this strong
broad plasmon peak had been well documented for
various Me- NPs, with sizes ranging all the way
from 2 to 100 nm. The microstructures and size of
the biosynthesized silver nanoparticles were studied
by TEM analysis. The typical TEM images of the
silver nanoparticles synthesized by rhizome extract
as reducing agent are shown in Fig. 2. The
micrograph shows formation of spherical like
morphology. The spherical like particles show very
small size about 5-10 nm.

100
Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103.
www.ijprsonline.com
ISSN: 2348 –0882
==============================================================================

2. Rai, M., Yadav, A., Gade, A., 2009. Silver
nanoparticles as a new generation of
antimicrobials. Biotechnol Adv 27, 76-83.
3. Elechiguerra, J,L., Morones, J,R., Burt, J.L.,
Camacho, A,B., Gao, X., 2005. The
bactericidal effect of silver nanoparticles
with HIV-1. J Nanobiotechnology 3, 6.
4. Naiwa, H.S., Hand Book of Nanostructural
Materials and Nanotechnology 2000.11, 1–
5p.
Conclusion
The silver nanoparticles synthesized using extracts
of rhizome samples was confirmed by color
changes and was characterized by UV-visible
spectrophotometer; the UV-visible spectra showed a
broad peak located at 422nm for silver
nanoparticles. The TEM analysis shows large
spherical shape particle with 200 nm size and a
small spherical like 5-10 nm size particles. This
technique has proved to be very useful for the
synthesis of nanoparticles from biological material.
Hence, we conclude that the synthesized
nanoparticles from C. longa more efficient due to
its biological origin and its smaller size and it can
be further analyzed for the usage in drug delivery
process and anti microbicidal properties.
4. References
1. Jianrong, C., Yuqing, M.,
Xiaohua,
W.,
Sijjiao,
Nanotechnology
and
BiotechnolAdv 22,505.

Nongyue, H.,
L.,
2004.
biosensors.

5. Song, J.Y., and Kim, B.S.,2008. Rapid
biologicalsynthesis of silver nanoparticles
using plant leafextracts. Bioprocess Biosyst
Eng. 6,313
6. Shankar, S,S., Ahmad, A., Sastry, M.,
2003.Geranium leaf assisted biosynthesis of
silver nanoparticles. Biotechnol Prog 19,
1627-1631.
7. Mariekie, G. and Anthony, P.,2006.
Microbial production of Gold nanoparticles,
Gold Bulletin, 39/1.
8. Mukherjee,P.,
Senapati,S.,
Mandal,D.,
Ahmad,A.,
Khan,M,I.,
Kumar,R.,Sastry,M.,2002.Extracellular
biosynthesis of bimetallic Au-Ag alloy
nanoparticles.Chem. Biochem, 3, 461-463
9. Langfield, R,D., Scarano, F,J., Heitzman,
M,E., Kondo, M., Hammond, G,B., 2004.
Use of a modified microplate bioassay
101
Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103.
www.ijprsonline.com
ISSN: 2348 –0882
==============================================================================

method to investigate antibacterial activity
in the Peruvian medicinal plant Peperomia
galioides. J Ethnopharmacol 94, 279-281.
10. Wiley, B,J., Xiong, Y., Li, Z,Y., Yin, Y.,
Xia, Y., 2006. Right bipyramids of silver: a
new shape derived from single twinned
seeds. Nano Lett 6,765-768.

11. Yamanaka, M. and Hara, K., 2005. A review
on the application of inorganic nanostructured materials in the modification of
textiles. J Appl Environ Microbiol 71,75897593.

102

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Biosynthesis Of Silver Nanoparticles Using Curcuma Longa And Their Antibacterial Activity

  • 1. Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== Biosynthesis Of Silver Nanoparticles Using Curcuma Longa And Their Antibacterial Activity S Elumalai1 and R Devika2 1 Associate professor, Dept. of Plant Biology & Plant Biotechnology, Presidency College, Chennai, India. 2 Research scholar, Dept. of Plant Biology & Plant Biotechnology, Presidency College, Chennai, India. Email: devinanotech25@gmail.com Abstract A rapid advance of nanotechnology has the potential approach for significant improvements in disease prevention, diagnosis and treatment. In this article, we report a simple and eco-friendly biosynthesis of silver nanoparticles (Ag-NPs) using silver nitrate as metal precursor in Curcuma longa. These Ag-NPs were characterized by UV–vis spectroscopy, and Transmission electron microscopy (TEM). These nanoparticles exhibited maximum absorbance in specific nano meter range in UV–vis spectroscopy. TEM micrographs revealed the formation of well-dispersed Ag-NPs with its size and morphology. Microbiology assay founds that Ag-NPs are effective against V.cholera bacteria. These developments raise exciting opportunities to diagnose and treat pathogenic mode of infection based on the various profiles to target diseases. Keywords: Silver nanoparticles, C. longa , Spectroscopic studies, Antibacterial Activity 1. Introduction A novel biosynthesis of nanoparticles is an exciting methods that have attracted significant attention due to their potential use in many applications, such as catalysis, drug delivery biosensor [1] antimicrobials and therapeutics [2,3]. New application of nanoparticles and nanomaterials are emerging rapidly [4]. Biological methods of nanoparticles synthesis using microorganism, enzyme, andplant or plant extract have been suggested aspossible ecofriendly alternatives to chemical andphysical methods [5]. As part of our work, we have observed that aqueous silver ions, when exposed to the rhizome extract of C. longa, are reduced in solution, thereby leading to the formation of an extremely stable silver particle. 2. Materials and Methods 2.1 Collection and Extract Preparation The rhizomes of C. longa were rinsed with fresh seawater and distilled water to remove associated debris. The cleaned material was then air dried to dryness in the shade at 30°C. The dried samples were finely powdered and stored at -20°C until use. Approximately 10 g of C. longa biomass was taken in a conical flask containing 100 mL of distilled water, kept for 24 hrs and then the aqueous solution components were separated by filtration. To this solution, AgNO3 (10-3 M) was added and kept for several hours at 24 hrs Periodically, aliquots of the reaction solution were removed and the absorptions were measured in a Elico UV-Vis spectrophotometer. 2.2 Synthesis and Characterization For the synthesis of Ag- NPs 1ml of rhizome extracts as test solution were incubated at room temperature for 1-2 hours. The silver nanoparticle solution thus obtained was purified by repeated 98
  • 2. Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== centrifugation at 15,000 rpm for 20min. Supernatant is discarded and the pellet is dissolved in deionised water. It is well known that Ag-NPs exhibit yellowish brown color in aqueous solution due to excitation of surface plasmon vibrations in Ag-NPs [6]. The silver nanoparticles were confirmed by colour changes and qualitatively characterized by UV-visible spectrophotometer on a Elico UV- Vis spectrophotometer. The bioreduction of Ag+ ions in solution was monitored using UV-Vis spectroscopyfrom zerotime reading was noted and double distilled water as blank, and incubated at culture condition. The samples were withdrawn at various time intervals and the absorbance was measured [7]. 2.3 UV-Vis spectroscopy analysis UV-Vis Spectral analysis was done by using HITACHI U-2900 Spetrophotometer. The UV-Vis Spectrophotometer analysis reveals the formation of silver nanoparticles by showing surface Plasmon resonance at 422 nm. UV-Vis Spectroscopy is one of the most widely used techniques for structural characterization of silver nanoparticles. The absorption spectrum of the brown silver colloids prepared by hydrazine reduction showed a surface Plasmon absorption band with a maximum of 422 nm indicating the presence of spherical or roughly spherical silver nanoparticles [8]. 2.4 TEManalysis of silver nanoparticles Sample for TEM analysis was prepared as mentioned in IR sample preparations. The sample was first sonicated (Vibronics VS 80) for 5 min. Silver nanoparticles was loaded on carbon-coated copper grids and solvent was allowed to evaporate under Infra light for 30 min. TEM measurements were performed on Phillips model CM 20 instrument operated at an accelerating voltage at 200 Kv. 2.5 Antibacterial activity: Bactericidal effects of Ag-NPs were studied against Vibrio cholera bacteria. Antimicrobial activity was demonstrated by modified method described [9]. Then 0.1 ml of the diluted microbial cultures was spread on sterile nutrient agar plate. The soaked and dried discs of 6 mm diameter of Whatman filter paper No: 1 were then placed on the seeded plates and gently pressed down to ensure contact [10]. Four replicates were placed for control, Ag-NPs, antibiotics and Ag-NPs combined with antibiotics Chloramphenicol, in each disc to confirm the inhibition a zone, and the plates were incubated at 37°C for 24 hours. After incubation period, the inhibition zone around the discs were measured and recorded, as the difference in diameter between the disc (6 mm) and growth free zone of V. cholera were measured. 2.6 Results & Discussion Silver nanoparticles were synthesized from AgNO3 solution containing Ag+ ions by treating with the rhizome extracts. The color of the solution changed to deep brownish color within 30 min of reaction with the Ag+ ions. The appearance of the deep brownish color indicated formation of silver nanoparticles. The formation of silver nanoparticles was confirmed by color changes followed by UVVis spectrophotometer analysis. It is generally recognized that UV-Vis spectroscopy could be used to examine size and shape-controlled nanoparticles in aqueous suspensions [11]. The UV-Vis spectrophotometer proved to be very useful technique for the analysis of some metal 99
  • 3. Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== nanoparticles. The UV- visible spectra (shown in Fig 1) indicated a strong Plasmon resonance that was located at ~422 nm. Presence of this strong broad plasmon peak had been well documented for various Me- NPs, with sizes ranging all the way from 2 to 100 nm. The microstructures and size of the biosynthesized silver nanoparticles were studied by TEM analysis. The typical TEM images of the silver nanoparticles synthesized by rhizome extract as reducing agent are shown in Fig. 2. The micrograph shows formation of spherical like morphology. The spherical like particles show very small size about 5-10 nm. 100
  • 4. Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== 2. Rai, M., Yadav, A., Gade, A., 2009. Silver nanoparticles as a new generation of antimicrobials. Biotechnol Adv 27, 76-83. 3. Elechiguerra, J,L., Morones, J,R., Burt, J.L., Camacho, A,B., Gao, X., 2005. The bactericidal effect of silver nanoparticles with HIV-1. J Nanobiotechnology 3, 6. 4. Naiwa, H.S., Hand Book of Nanostructural Materials and Nanotechnology 2000.11, 1– 5p. Conclusion The silver nanoparticles synthesized using extracts of rhizome samples was confirmed by color changes and was characterized by UV-visible spectrophotometer; the UV-visible spectra showed a broad peak located at 422nm for silver nanoparticles. The TEM analysis shows large spherical shape particle with 200 nm size and a small spherical like 5-10 nm size particles. This technique has proved to be very useful for the synthesis of nanoparticles from biological material. Hence, we conclude that the synthesized nanoparticles from C. longa more efficient due to its biological origin and its smaller size and it can be further analyzed for the usage in drug delivery process and anti microbicidal properties. 4. References 1. Jianrong, C., Yuqing, M., Xiaohua, W., Sijjiao, Nanotechnology and BiotechnolAdv 22,505. Nongyue, H., L., 2004. biosensors. 5. Song, J.Y., and Kim, B.S.,2008. Rapid biologicalsynthesis of silver nanoparticles using plant leafextracts. Bioprocess Biosyst Eng. 6,313 6. Shankar, S,S., Ahmad, A., Sastry, M., 2003.Geranium leaf assisted biosynthesis of silver nanoparticles. Biotechnol Prog 19, 1627-1631. 7. Mariekie, G. and Anthony, P.,2006. Microbial production of Gold nanoparticles, Gold Bulletin, 39/1. 8. Mukherjee,P., Senapati,S., Mandal,D., Ahmad,A., Khan,M,I., Kumar,R.,Sastry,M.,2002.Extracellular biosynthesis of bimetallic Au-Ag alloy nanoparticles.Chem. Biochem, 3, 461-463 9. Langfield, R,D., Scarano, F,J., Heitzman, M,E., Kondo, M., Hammond, G,B., 2004. Use of a modified microplate bioassay 101
  • 5. Int. J. Pharm. Res. Sci., 2014, 02(1), 98-103. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== method to investigate antibacterial activity in the Peruvian medicinal plant Peperomia galioides. J Ethnopharmacol 94, 279-281. 10. Wiley, B,J., Xiong, Y., Li, Z,Y., Yin, Y., Xia, Y., 2006. Right bipyramids of silver: a new shape derived from single twinned seeds. Nano Lett 6,765-768. 11. Yamanaka, M. and Hara, K., 2005. A review on the application of inorganic nanostructured materials in the modification of textiles. J Appl Environ Microbiol 71,75897593. 102