INTRODUCTION: Nanotechnology refers to a field of applied science and technology
whose theme is the control of matter on the atomic and molecular scale, generally 100
nanometres or smaller, and the fabrication of devices or materials that lie within that size
range.
CNTs are allotropes of carbon with a nanostructure that can have a length-to-diameter ratio
of up to 28,000,000:1, which is significantly larger in comparison to any other material.
In the nanotechnology development, CNTs have attracted a great deal of attention due to their
unique structural, electrical and mechanical properties. Some of the properties ofCNTs for
like superior strength, flexibility, electrical conductivity ·andeasy chemical fictionalization
have been exploited for the biological application both at the cellular and molecular level.
Amino acid -metal complex: Proteins are bio polymers comprising of amino acids as
monomer. Amino acid~ are compounds containing -NH2 and COOH groups within a single
molecule. NH2 group possess one lone pair of electron on nitrogen atom. With the help of
these groups amino acids form complexes with metals. In these complexes different chains of
amino acids are clubbed together. These compounds on decomposition give carbon -metal
Nanotubes. When aqueous solution of Nickel Salt is allowed to react with amino acids
present in egg albumin the lone pair present on nitrogen of -NH2 and oxygen of COO- of
COOH group present in amino acid form complex with Ni (ii). In this way nickel (ii) forms
cross link between two amino acid chains.
The structure of amino acids present in egg albumin is very complex and it is very difficult to
produce exact structure of amino acid-metal complex. Egg albumins contain a number of
different chemical compounds, which also form complex with metal. This metal -amino
acids complexes are heated in muffle furnaGe till decomposition. Theses complexes on
decomposition give carbon Nanotubes.
Method of Preparation of CNTs Using Egg and Nickel Chloride in Aqueous
Medium
Preparation ofCNTs is a two step process in first step amino acid -metal complex is prepared
and in second step they are decomposed in muffle furnace.
Preparation of EggNiCh (Aq)
1CNTs are fonned only when the concentration of carbon at a particular point is
high. So, to club various protein molecules together, aqueous solution of Nickel Salt is
allowed to react with amino acids present in Egg. The lone pair present on nitrogen of -NH2
and COO· of COOH group present in amino acid fonn complex with Ni (ii). In this way
nickel (ii) fonns cross link between two amino acid chains.
First of all Egg yolk is separated from egg serum and 2gms of it is kept in glass
beaker. Now 5ml of IN solution ofNiCh is prepared in water and is mixed with egg yolk for
the reaction. The reaction mixture is kept undisturbed for 15 days· for precipitation and drying
of complex. After 15 days jelly like green complex is fonned. The complex is washed with
water and dried again. The weight of dry complex is 2.457gms. The dried complex is now
ready to decompose and for further studies.
Decomposition
The dried complex is decomposed at different temperatures 800°C and 900°C in a
muffle furnace. For decomposition 2gmof complex is weighed and kept in crucible. The
complex is then kept in muffle furnace for ten minutes at 800°C. The weight of decomposed
complex is O.l57gms.The same process is repeated at 900. The weights of complex is
O.l53gms.
The complex of amino Acid and metal is expected to contain following groups:
(i) NH2 group .
(ii) COOH group
(iii) CH2 group
(iv) Amide group etc.
The presence of these groups is confinned by using IR and NMR studies.
.Characterization of amino acid -metal complex:
IR Spectroscopy
The IR spectra of compound prepared by the reaction of Nickel salt and Egg
protein in aqueous is shown in Fig 1.1
2pj,
..."\}
Ii
2>
M
D"Hi
mn
tt ,r .,
u
., )I) I. ~l
Fig: 1.1 IR Spectra ofNickel salt and egg protein in aqueous medium
The results of IR spectra of Nickel salt and egg protein in aqueous medium are
given below:
1
1. Functional group C-O stretching is found at an absorption band of 1376 cm-1.
2. Functional group N-H stretching (symmetric and asymmetric) is found at an
.absorption band of 3369 cm-1.
33. Functional group -oH stretching in -COOH is found at a absorption band of
2925.78 cm-
I
. It can be because of C-H stretching in alkanes.
4. Functional group -oH stretching (superimposed upon -CH stretching) is found at
an absorption band of2854 cm-
I
. It can be because ofC-H stretching in alkanes'.
5. Functional group c=o (carbonyl group) linked to the carbon which is attached to
-NHz group found at a absorption band of 1744, 1774,1794 cm-I.
6. Functional group c=o stretching in -COOH is found at an absorption band of
1630 cm-
I
.
7. Functional group C-N stretching in amines is found at an absorption band of 1092
& 1167cm'l. .
8. NH stretching in NH 3 + in Zwitter ionic form is found at3007cm-
1
NMR Spectroscopy
The NMR spectra of compound prepared by the reaction of Nickel salt and Egg
protein in aqueous medium is shown in Fig 1.2:
4WO,,"'''''''''_
,
ca,~
(;Ii
1$
I I
(CHJCO CY'....
• I I I
~ •OiQQI!1I let;1so
Oq)l (~PH
- tlQ
~-C'·N,•.•
,-CI'.,'!-•.
-COu)----- C,CII.II·-
,CCli:O • i t::{;M~
~kGQ\c-- I ·eijQ4f,o
N~N -CCCtt,'- riC":
I
Cfl.ct.
·;.,J.~ , f
- I I
11.3 J+¥ Cr,COOH 1"-.(ill ~f\tb1 1Y4nt~
C.M Mt
1--··-1'----·
j ,Ii," ,
Fig: 1.2 NMR Spectra ofNickel salt and egg protein in aqueous medium
The results ofNMR spectra ofNickel salt .and egg protein in aqueous medium are
given below: 1
1. For (CH3)3C- the chemical shift (0) value is 1.45 ppm.
2. For CH2- and NH2 the chemical shift (0) value is 4.4 ppm.
3; For -CH- the chemical shift (0) value is 5.5 ppm.
4. For -COOH the chemical shift (0) value is 6 -7.8 ppm.
5From the results, ofNMRand IR it was found that the compound contain all the
above said groups. In other words, we can say that the desired chemical compound is formed.
Using IR and NMR data we can deduce the structure of amino acid. The structure
is drown in the Fig1.3
HO
Fig1.3 Structure deduced by NMR &IR spectra
This structure resemble with Lucien protein which is an essential constituent.of
egg albumin and all the groups identified in IR and NMR spectra are present in the structure.
From the result ofNMR and IR it is clear that +1 effect plays a vital role in the
p~eparation of protein nickel complex, the amino acid more no. of alkyl group form
compound with nickel. Chemical and physical evidences suggest that alkyl group, relative to
hydrogen atoms release electrons to carbon. If an alkyl group is attached to an electron
deficient carbon it donates electrons through the carbon-carbon a bonds.
2
The structure of r"ucien Ni
2
+ complex is given in the Fig 1.4
Fig. I.4Structure ofLucien Ni
2
+ complex
6Characterisation of study of properties of. CNTs: These CNTs are under process of
characterization using scanning probe instruments like DLS, SEM, XRD, etc. Their
properties were studied using VSM,TGA, DTA,DSC.
. Characterization of.CNTs Using DLS
CNTs were dissolved in ethylene glycol and analyzed.
Result of CNTs of Complex Egg NiCh (800Aq)
DLS Graph of CNTs prepared by using egg protein and nickel salt in aqueous
medium and by decomposing the compound in muffle furnace at 800°C is shown in Fig 15
7SIze Distribution Report by intensity
t¢.
~~
......_~W)
IOP_ ..... I..._
..........:
~
Malvern
......... ". ..............i., ee,
~_.'''''Ge,td
........ "at
_millltllil")ttt.ma
....... 11.• .,.....~ ......,....07••tOU:&"
---T rq; 2"UJ UaIlIM: to
c:o.t : ..,. _411 ~..
c. u; ;·'1 .
l""'.~" $t1f,~ t;
..... ~- 1:
. _ ...ft: C,. ~:
.......~:.~......,.....
.......
40B
iI,.
0.000
,,~
•• ...
00
.... 710.
14"
eooo
•••"" •• If••• • oj. , •.~ _ : •.-. "' -It - •••~fi; (1;-41),•• If •• If: •••••"' :
v. v"
~ ., ..
·
· " -,.- ,. .<
" - "', - - '" - '" ,.
I· :. -"' .•.•., ..•...••r·'········,· ,;,.:.:..'" .."' ...•.·t···.. ·· -.' -; ...- ".it••" ... :
f .J J ; .L i.~....... ..~ ~ ~ :
·
. ..-.---.....----.....--- Cl:'
Fig 1.5 DLS Graph ofCNTs prepared by complex ofEggNich (800Aq)
8Table:1 DLS Data
S.No. Type Sample Temp. z- Pdl Pkl Pk2 Pk3 Pkl Pk2 Pk3 Mean Multimodal Cumulants Number Volume
Name °C Ave Mean Mean Mean Area Area Area Count Fir error Fit Error Mean Mean
Int Int Int Int .Int Intensity Rate
°C d.nm d.nm d.nm d.n~ % % % kcps d.nm d.nm
1. Size 800c WI 25 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 540.9 0.00 0.00 0.00 0.00
2. Size 800c WI 25 193.1 1.00 4093 59.56 6428 53.6 35.0 11.4 232.1 0.00579 0.00179 56.20 2405
3. Size 800c W2 25 239.6 1.00 811.2 49.66 5179 58.3 29.1 12.6 257.4 0.00420 0.00575 45.95- 1045
4. Size 800c W3 24.9 813.0 1.00 4615 113.8 0.000 85.4 '14.6 0.0 1036.4 0.00604 0.00303 107.4 4546
9Discussion ofDLS Results
DLS gives us idea about the particle size. It is based on the fact larger the particle
size more light will be scattered by it. It is clear from the graph that when the metal protein
complex were decomposed at 800°C, the number of particles having nanodimensions (from
10 to 100nm) is more, as the from the graphs it is clear that the peaks corresponding to
particle size 10 to1OOnm are smaller than the peaks corresponding to 900 to 10000nm. This
shows high concentration of smaller particles as small particles scatter less amount of light.
O.Truspike' Vych et al
3
characterized Carbon Nanotubes Thermo Tropic
pneumatic Liquid Crystal Composites, using DLS and reported similar findings. Therefore,
DLS result conform the formation ofnanosized particles
Characterization Using SEM
SEM results give us idea about surface topography ofCNTs. It also gives us idea
about their surface morphology. The CNTs were analyzed at Nano centre of Allahabad
University.
SEM Images of CNTs of Complex Egg NiCh (800 Aq)
SEM images' of CNTs formed by the reaction of egg protein and nickel salt in aqueous
medium and decomposed at 800°C in muffle furnace. This image is shown in Fig 1.6.
The figure shows form~tion ofCNTs
4
,5,6
10Fig.I.6 SEM Images ofCNTs of complex Egg NiCh( 800Aq)
O.Truspike Vych etal
3
characterized Carbon Nanotubes Thermo Tropic
pneumatic Liquid Crystal Composites using ESEM the SEM figure resembles with the figure
ofmy CNT samples And conforms the formation ofCNTs
K.Atre etal
7
.of Nanomaterial and nanoresearch laboratory, USA synthesized
vertically aligned carbon nanotubes and characterized using SEM. Some of their SEM
images showed resemblance with our SEM images. This conform the formation of
CNTs.
Results of Sample Analyzed by Using XRD
Different samples ofCNTs were analyzed using XRD. The analyses were carried
out at lIT Roorkee. The results are given under:
XRD Graph of Complex Egg NiCh (900Aq)
The CNTs were prepared by using egg protein" and nickel salt in aqueous medium
and decomposed in muffle furnace at 9000
C.
The graph ofXRD analyses ofthe CNTs is shown in Fig 1.7 and data is given in table 2.
11Table 2: XRD data
20An2le Intensity d-value
11.261 470 d=7.85143
20.806 481 d=4.26593
24.118 440 d=3.68708
28.378 645 d=3.14258
29.426 467 d=3.03292
31.142 442 d=2.86963
37.32- 431 d=2.40752,
40.542 498 d=2.22334
43.357 448 d=2.08528
44.492 626 d=2.03467
50.184 400 d=1.81643
51.812 453 d=1.76311
73.684 350 d=I.28467
76.27 373 d=1.24742
91.199 338 d=1.07815
94.064 344 d=1.05271
XRDGraph 1000
900
800
> 700
:s 600
; 500
c400
- 300
200
100
o
o 20 40 60
2 ..1htt.scale
so 100
Fig: 1.7 XRD Graph of complex Egg NiCh (900Aq)
12The XRD results were found to be almost similar to the results reported by for multi walled
CNTs prepared by C.~.Bharadwalby pyrolysis of turpentine oil (29=25.6) and with loan
Stalin etal
9
,who prepared CNTs by catalytic pyrolysis of phenol formaldehyde resin(29
=26.2).
Characterization using TEM
TEM image of CNTs is shown in fig 1.8. The image of CNT is on magnification
scale of 100nm.
These images shows resemblance with the results ofChang Xin Chen, Wenzhe Chen
Yafei Zhang13
and LI Wei-xue, HAO Yuan" CUI Yong-fu, DAI Jian-feng" CHEN
Ti-jun, WANG Qing, , WU Zhong-li, LI Yang14
•
Fig.l.8 TEM image ofCNTs
13Study of Properties ofCNTs
With synthesis and characterization of CNTs the study of properties is also
equally important. So some major properties ofCNTs are studied, they are as under:
TGA, DTA, DTG Thermogram of CNTs of Complex Egg NiCh (950Aq)
TGA, DTA, DTG thermo gram ofCNTs prepared by the reaction ofegg protein and Nickel
salt in aqueous medium and decomposing the compound at 950°C.The analyses was carried
out by adjusting the furnace temperature at 11 OO°C.The thermogram is shown in Fig 1.9
14-150.0
neel
0.09 mg/min
////
352 eel
'204 ug/min
400 eel
68.0% /
, 160.0
--' 140.0
120.0
--100.0 ....
~
·····80.0
1043 eeT 60.0
40.8 %
\ ---i
!40.0
1000 eel
41.9 % .
020
0.00
-0.40 c
I;'
l:h
E
o
-G.60 b
-080
100 200 300 400 500 600 700 800 900 1000
Temp eel
Fig: 1.9 TGA, DTA, DTG Thermo gram ofCNTs of complex EggNiCh(900Aq)
Discussion of,TGA, DTA, DTG Thermogram ofCNTs ofComplex Egg NiCh (900Aq)
The CNTs were prepared using egg protein and nickel salt in aqueous medium
and decomposing the result and compound in muffle furnace at 900°C. At the time ofTGA,
DTA, DTG analysis the temperature of the furnace was adjusted to 1100°C. From the thermo
gram it is clear that dehydration takes place between the temperature ranges from 300 to
500°C. This is also confirmed by weight losses in DTA, DTG curves. The temperature of the
source reached up to 1043°C. At this temperature 40.8% sample was found to be un-
decomposed. The melting point was not achieved till this temperature.
Chang etal
lO studied thermal properties ofmultiwalled carbon nanotubes dispersed in
water and associated with biological effect using TGA and DTA.
Sumio Iijima etal!! also studied the thermal properties of carbon nanotubes
ultrasonicated in mono~hloro benzene using TGA.
Their thermo grams showed similarities with the thermo grams of my sample but
there CNTs decomposed at about 800 to 850°C.They carried out the test in air but my
samples were decomposed in the atmosphere of Nitrogen. That might be the reason of such
difference..
Study ofMagnetic Properties of CNTs
15Magnetic properties of CNTs are studied using VSM. The magnetic movement of
CNTs is compared with the blank. The samples were analyzed at lIT Roorkee.
VSM Graph' of Sample ofComplex Egg NiC12 (900Aq)
Graph ofMagnetic properties ofCNTs prepared by using Nickel salt and egg protein in
aqueous medium and by decomposing the compound at 900°C. The graph of blank is also
gwen for reference. The graph is shown in Fig 1.10 and graph of blank reference is given in
Fig l.ll
Table3:VSM Data(Sample)
Magnetic Magnetic
Field moment
9664 0.469
7466 0.458
7288 0.045
5435 0.0435
4488 0.042
4097 0.0408
4072 0.0397
3893 0.0393
3684 0.0387
3376 0.0344
2747 0.0328
2540 0.0321
1598 0.0297
368 0.0223
-4 -0.0009
-423 -0.0075
-843 -0.0127
-1261 -0.0103
-1777 -0.0126
-2195 -0.0104
16.----------------------------------------~-
0.6
0.5
,,~".- .._.~-_.~--~.~ ..-..,..,
l
I
I~ 141
1
8
I:E u
~ c
:ll'
I:E I·
I
I
I -4000
I
-2000
0.4
0.3
0.2
0.1
-0.1
2000 4000 6000
Magnetic Field
8000 10000 12000
Fig: 1.10 VSM Graph of Sample ofcomplex Egg NiCh (900Aq)
Table4:VSM Data(B1ank)
Magnetic Magnetic
field moment
9601 0.00375
8989 0.003711
178988 0.003683
8871 0.003623
8746 0.003514
7665 0.003458
7499 0.003341
3700 0.003294
3178 0.003212
2971 0.003195
2760 0.003045
2337 0.003033
2130 0.002861
1918 0.002501
1881 0.00197
-1265 0.001381
-2203 0.001327
-2623 0.001007
-3042· 0.000987
-4396 0.000742
0.004
0.0035
0.003
"l::
0.0025
I
Q/
vJ
E 0
:::iE
0.002 .lol
-:;:
I:
~ :::iE
~ 0.001
0.0005
_ .............. .............................................. & .....
-6000 -4000 -2000 0 2000 4000 6000 SOOO 10000 12000
I'IiI"gnetie Field
Fig: 1.11 VSM Graph ofBlank
LI Wei~Xue etal
12
studied the magnetic properties of multi-walled carbon nanotubes
encapsulated Fe/Co particles. There graph shows resemblance with the graph of my sample.
In both the graphs CNTs are paramagnetic in nature. Graphs show increase in magnetic
moment with increase in magnetic field strength.
18K.Atre etaf of Nanomaterial and mino research laboratory, USA synthesized
vertically aligned carbon nanotubes. They also studied magnetic properties using VSM.
There VSM graph also showed similar findings and resembled with the graph ofmy sample.
Conclusion
CNTs can be prepared by the reaction of Amino Acid with Nickel salt and by
decomposing the compound formed. Chemical compound can be formed in both aqueous and
organic medium. This shows that even after denaturation protein formed desired compound
with Nickel salt. This indicates that primary peptide structure is involved in compound
formation.
De~omposition temperature also plays a vital role in the formation of CNTs.
DLS and SEM results reveals that 800 and 900 are the optimum temperatures for the
. .
formation ofCNTs. SEM and DLS results also confirm the form,ation ofCNTs.
The formation of CNTs is again confirmed by XRD results. XRD results reveal
that MWNT are formed.
Some properties of CNTs were also studied TGA, DTG, and DTA results give
idea about thermal properties of CNTs. These studies tell us that the CNTs possess hi'gh
thermal stability and high melting point.
VSM studies confirm the paramagnetic nature ofCNTs.
Refere~ce:
1. B.DMistry, A Handbook of Spectroscopic Data Chemistry ,2009 pp27-
53&101-127
2. Organic Chemistry ,Alans, Wingrove, Robert.L.Carot pp63
3. O. Truspike Vych, N Callings, T. Hasan, V. Scardaci, AC Ferrari and co-
workers ,Characterization of Carbon Nanotubes Thermo Tropic Pneumatic
Liquid Crystal Composites, Journal ofPhysics Vol. 41 .2008.
4. B. Tiwari, I.P. Tripathi, Sanjay Saxena, Synthesis and Characterization'of
CNTs by Prepared by Denatured Protein, Inventi International Journal, Vol. 1 ,
2011.
5. B. Tiwari, I.P.Tripathi, Sanjay Saxena, Synthesis
. Carbon metal Nanotubes, Conference Proceedings
Physics, 2010.
and Characterization of
American Institute of
6. B. Tiwari, I.P. Tripathi and Sanjay Saxena, Synthesis of Carbon Nanotubes
Using Sp'inach, Characterization and Study ofMagnetic Properties. Journal of
Indian Chemical Society, Vo1.89, ppI143-1148, 2012.
197. K.Aatre et ai, Synthesis ofVertically Aligned Carbon Nanotube and Magnetic
CNTs For Cellular Growth and Detection, Proc ofSPIC, Vol693I, 2008.
8. C.R. Bhattacharji, A.Nath and coworker ,Synthesis and Characterization of
Carbon Nanotubes using a national precursor turpentine oil ,Science Journal
UBU vol.2, pp36-42,2011
9. loan Stalin Etal, Synthesis and Characterization of Carbon Nanotubes by
Catalytic Pyrolysis of Phenol formal resigns, Physics vo1.37, pp44-48,2007 .
10. X.Chang et aI, Characterization of Multiwalled Carbon Nanotubes Dispersing
in Water and Associate With Biological Effects, Journal of Nanomaterials
vo1.2011,2011
II. Sumio Iijima et aI, Thermo gravimetric analysis of single wall carbon
. nanotubes ultrasonicated in mono chIaro benzene, Chemical Physics
letters,vo1364 pp420-426,2004
12.L~ Wei-xue, HAG Yuan, , CUI Yong-fu, DAI Jian-feng, , CHEN Ti-jun,
WANG Qing" WU Zhong-Ii" LI Yang, Magnetic.Properties ofMulti-Walled
Carbon Nanotubes Encapsulated Fe/Co Particles, Journal ofTrans Non Ferrous
Matal Society of china Volume 17 pp. 696-699 , 2007.
I3.Chang Xin Chen, Wenzhe Chen Yafei Zhang, Synthesis ofCarbon Nanotubes by
Pulsed Laser Ablation at Normal Pressure in Metal Nano-Sol., Material Synthesis
Papers, Vol. 28 Issue 2, pp. 121-127,2005.
14.LI Wei-xue, HAG Yuan" CUI Yong-fu, DAI Jian-feng" CHEN Ti-jun, WANG
Qing, , WU Zhong-Ii , LI Yang, Magnetic properties of multi-walled carbon
nanotubes encapsulated with Fe/Co, Journal of Trans Non FerrousMetal Society of
China Vol. 17 pp. 696-699, 2007.