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!!Om Sai Ram!!
Volume-2, Number- 1, January-April, 2011
ijCEPr www.ijcepr.com
International Journal of
Chemical,Chemical,Chemical,Chemical,
Environmental andEnvironmental andEnvironmental andEnvironmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Editor-in-Chief
Prof. (Dr.) Sanjay K. Sharma
STATUTORY WARNING
Articles, Data, Figures, Scientific Content and its Interpretation and Authenticity reported by author(s) and
published in ijCEPr are the exclusive views of author(s). The Editorial board, ijCEPr is not responsible for
any controversies arising out of them.
Published by:
23, ‘Anukampa’, Janakpuri, Opp. Heerapura Power Station,
Ajmer Road, Jaipur (India)
Phone: 0091-141-2810628, 09414202678
E-mail: [email protected]
Website: www.ijcepr.com
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i
ijCEPr www.ijcepr.com
International Journal of Chemical,Chemical,Chemical,Chemical, Environmental andEnvironmental andEnvironmental andEnvironmental and Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Volume-2, Number-1, January-April, 1-66 (2011)
Contents…
CHEMICAL RESEARCH
Synthesis of 2-Substituted-1,3,4-Oxadiazole Derivatives
Vijay V. Dabholkar and Nitin V. Bhusari
1-4
Molecular Interaction Studies between H-Bonded Ternary Mixtures of p-Cresol with simple
Aldehydes in Cyclohexane at Different Temperatures M. Aravinthraj, S. Venkatesan
and M. Kamaraj
5-11
Chemical Analysis on Different Oils Use in Tyre Tread Cap Compound
N. Kumar, R.K. Khandelwal, P.L. Meena, K. S. Meena, T.K. Chaki, D.K. Mahla and S. Dasgupta
12-19
Infra Red Spectral and X-ray Diffraction Study of Fe (II), Co(II), Cu (II), Metal Chelates
with N1-(5, 6-dimethoxypyrimidin-4-yl) Sulphanilamide
Jitendra H. Deshmukh and M. N. Deshpande
20-25
Comparative Study between soda Lignin and soda Anthraquinone lignin in terms of
Physiochemical Properties of Ipomoea carena Preeti Nandkumar
26-29
ENVIRONMENTAL RESEARCH
Use of Millet Husk as a Biosorbent for the Removal of chromium and Manganese Ions from
the Aqueous Solutions.
Manju Chaudhary
30-33
Study Regarding Lake Water Pollution with Heavy Metals in Nagpur City (India)
P.J. Puri, M.K.N. Yenkie, S. P. Sangal, N.V. Gandhare, G. B. Sarote
34-39
Knowledge, Attitude and Practices regarding Waste Management in Selected Hostel
Students of University of Rajasthan, Jaipur Lalita Arora and Sunita Agarwal
40-43
Utilisation of Thiocyanate (SCN-) by a Metabolically Active Bacterial Consortium as the Sole
Source of Cellular Nitrogen
Yogesh B. Patil
44-48
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ii
Construction of an open loop temperature control system for thin film fabrication in PC
based instrumentation.
K.Tamilselvan* , K.Anuradha, S.Deepa , O.N.Balasundaram and S.Palaniswamy
49-51
PHARMACEUTICAL RESEARCH
Accumulation of Natural Antioxidants in Ferns Exposed to Mutagenic Stress
Alok Kr. Singh, Santosh Kr. Singh, Satish K. Verma, H.V. Singh, A.K. Mishra, Pavan K.
Agrawal, Abhishek Mathur and Md. Aslam Siddiqui
52-55
Reversed Phase HPLC Analysis of Valsartan in Pharmaceutical Dosage Forms V. Bhaskara Raju and A. Lakshmana Rao
56-60
A Review on Fibrinolytic Enzyme: Nattokinase
Haritha Meruvu and Meena Vangalapati
61-66
INDEX of Contributors of this issue
Authors Guidelines: for RASAYAN J. Chem.
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iii
ijCEPr www.ijcepr.com
International Journal of Chemical, Environmental and Pharmaceutical ResearchChemical, Environmental and Pharmaceutical ResearchChemical, Environmental and Pharmaceutical ResearchChemical, Environmental and Pharmaceutical Research
Volume-2, Number-1, January-April, 1-66 (2011)
AUTHOR INDEX OF THIS ISSUE
IJCEPR widely covers all fields of Chemical, Environmental and Pharmaceutical Research.
Manuscript Categories: Full-length paper, Review Articles, Short/Rapid Communications.
Manuscripts should be addressed to:
Prof. (Dr.) Sanjay K. Sharma
Editor-in-Chief
23, ‘Anukampa’,Janakpuri, Opp. Heerapura Power Station,
Ajmer Road, Jaipur-302024 (India)
E-mail: [email protected]
Phone:0141-2810628(O), 09414202678(M)
A. Lakshmana Rao,56
A.K. Mishra,52
Abhishek Mathur,52
Alok Kr. Singh,52
D.K. Mahla,12
G. B. Sarote,34
H.V. Singh,52
Haritha Meruvu,61
Jitendra H. Deshmukh,20
K. S. Meena,12
K.Anuradha,49
K.Tamilselvan,49
Lalita Arora,40
M. Aravinthraj,5
M. Kamaraj,5
M. N. Deshpande,20
M.K.N. Yenkie,34
Manju Chaudhary,30
Md. Aslam Siddiqui,52
Meena Vangalapati,61 N. Kumar,12
N.V. Gandhare,34
Nitin V. Bhusari,1
O.N.Balasundaram,49
P.J. Puri,34
P.L. Meena,12
Pavan K. Agrawal,52
Preeti Nandkumar,26
R.K. Khandelwal,12
S. Dasgupta,12
S. P. Sangal,34
S. Venkatesan,5
S.Deepa,49
S.Palaniswamy,49
Santosh Kr. Singh,52
Satish K. Verma,52
Sunita Agarwal,40
T.K. Chaki,12
V. Bhaskara Raju,56
Vijay V. Dabholkar,1
Yogesh B. Patil,44
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical Pharmaceutical Pharmaceutical Pharmaceutical ResearchResearchResearchResearch
Vol. 2, No.1, 1-4
January-April, 2011
Vijay V. Dabholkar and Nitin V. Bhusari
Synthesis of 2-Substituted-1,3,4-Oxadiazole Derivatives
Vijay V. Dabholkar* and Nitin V. Bhusari
Organic Research Laboratory, Department of Chemistry, KC College, Churchgate, Mumbai-20 1Mumbai University, Maharashtra, Mumbai (India)
*E-mail: [email protected] Article History:
Received: 30 December 2010
Accepted: 20 January 2011
ABSTRACT Diethyladipate on reaction with Hydrazine hydrate gave Succinohydrazide (1) which on further treatment with Carbon disulfide,
Aromatic aldehydes and Cynogen bromide yielded 1,2[di-(2-Mercapto-1,3,4-oxadiazole-5yl)] ethane (2), 1,2[di-(2-Phenyl-1,3,4-
oxadiazole-5yl)] ethane (3a-f) and 1,2[di-(2-Amino-1,3,4-oxadiazole-5 yl)] ethane (4) respectively. The structures of the
compounds have been elucidated on the basis of spectral analysis.
Keywords: Diethyladipate, Succinohydrazide, Carbon disulfide and Cynogen bromide. ©2011 ijCEPr. All rights reserved
INTRODUCTION The chemistry of heterocyclic compounds has been an interesting field of study for a long time. The synthesis of
novel Oxadiazole derivatives and investigation of their chemical and biological behavior have gained more
importance in recent decades for biological, medical and agricultural reasons. Different classes of Oxadiazole
compounds possess an extensive spectrum of pharmacological activities. In particular, compounds bearing 1,3,4-
Oxadiazole nucleus are known to exhibit unique anti-edema and anti-inflammatory activity [1-4,9]. Differently
substituted Oxadiazole moiety has also been found to have other important activities such as analgesic [3,4]
antimicrobial [5,6], antimycobacterial [7], anticonvulsant [8], antitumor [9], antimalarial [10] and anti-hepatitis B
viral activities [11]. Substituted 1,3,4-Oxadiazoles exhibit antibacterial [12], Pesticidal [13] and antifungal [14]
activities.
1,3,4-oxadiazoles are biologically active [15], synthetically useful and important heterocyclic compounds. for these
reasons the chemistry of 1,3,4-oxadiazoles have been the subject of many investigations [16-19].One pot synthesis
of 1,3,4-oxadiazoles by the reaction of appropriate hydrazide and carboxylic acid has been reported [20]. Cerric
ammonium nitrate has received considerable attention as an inexpensive and easily available catalyst for various
organic reactions such as Oxidation, Oxidative addition, Nitration, Photo-oxidation, Polymerization etc. In recent
report Cerric ammonium mediated synthesis of 1,3,4-Oxadiazoles has also been described [21,22].
MATERIALS AND METHODS Melting points of all synthesized compounds were determined in open capillary tubes on an electrothermal apparatus
and are uncorrected. The progress of reaction was monitored by thin layer chromatography on silica gel coated
aluminium plates (Merck) as adsorbent and UV light as visualizing agent. IR spectra (KBr in cm-1
) were recorded on
a Perkin-Elmer spectrophotometer in the range of 4000-400 cm-1. 1H NMR spectra were recorded on a Varian 500
MHz NMR spectrometer using CDCl3/DMSO-d6 as solvent and TMS as an internal standard (chemical shifts in
δppm).
Succinohydrazide (1): General procedure
A mixture of Diethyladipate (0.08 mole), Hydrazine hydrate (3.85 ml, 0.08 mole), and ethanol (10 ml) was refluxed
on water bath for 4-5 hrs. The mixture becomes almost solid. This reaction mixture is allowed to cool at room
temperature. The white solid so obtained is then filtered and washed with cold ethanol and finally recrystallized
from hot water to yield (1).
1,2[di-(2-Mercapto-1,3,4-oxadiazole-5yl)] ethane (2)
A mixture of Succinohydrazide (1.0 gm, 0.0068 mole), Carbon disulfide (0.82 ml, 0.0136 mole), and Potassium
hydroxide (0.768 gm, 0.014 mole) was refluxed in ethanol (10 ml) on water bath for 5-6 hrs. The reaction is then
allowed to cool. The red colored solid so obtained is then filtered and washed with ethanol and finally recrystallized
from hot dichloromethane to yield (2).
1,2[di-(2-Phenyl-1,3,4-oxadiazole-5yl)] ethane (3a-f)
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Vol.2, No.1, 1-4 (2011)
Vijay V. Dabholkar and Nitin V. Bhusari 2
A mixture of Succinohydrazide (1.0 gm, 0.0068 mole), aromatic aldehydes (0.0136 mole), and Cerric ammonium
sulphate (0.5 gm, 0.0008 mole), in Dichloromethane (10 ml) as a solvent was taken in 100 ml round bottom flask
and the mixture was refluxed on water bath for 4-5 hrs. After monitoring the reaction on TLC, the reaction mixture
was cooled and dumped on to the ice, filtered and recrystallized from ethanol.
1,2[di-(2-Amino-1,3,4-oxadiazole-5 yl)] ethane (4) A mixture of Succinohydrazide (1.0 gm, 0.0068 mole), Cynogen bromide (0.0136 mole), and Sodium bicarbonate (1
g, 0.012 mole), in ethanol (10 ml) as a solvent was taken in 100 mL round bottom flask and the mixture was
refluxed on water bath for 4-5 hrs. After monitoring the reaction on TLC, the reaction mixture was cooled and
dumped on to the ice, filtered and recrystallized from dimethylformamide to yield (4).
The schematic data of the compound 2, 3(a-f) and 4 are listed in the Table-1.
Antimicrobial evaluation
Representative samples were screened for their antimicrobial and antifungal activity against gram-negative bacteria,
E coli and P aeruginosa and gram-positive bacteria, S aureus, and C diphtheriae using disc diffusion method [23,24].
The zone of inhibition was measured in mm and the activity was compared with standard drug. The results of
antibacterial screening studies are reported in Table-2.
RESULTS AND DISCUSSION Diethyladipate on reaction with Hydrazine hydrate gave Succinohydrazide (1) which on further treatment with
Carbon disulfide, Aromatic aldehydes and Cynogen bromide yielded 1,2[di-(2-Mercapto-1,3,4-oxadiazole-5yl)]
ethane (2), 1,2[di-(2-Phenyl-1,3,4-oxadiazole-5yl)] ethane (3a-f) and 1,2[di-(2-Amino-1,3,4-oxadiazole-5 yl)]
ethane (4) respectively with good yield.
Further, the representative compounds were screened for their antimicrobial activity against gram negative as well
as gram positive bacteria, which shows convincing activity.
ACKNOWLEDGEMENTS The authors are grateful to the Principal Ms. Manju J. Nichani and Management of K.C. College, Mumbai for
providing necessary facilities. Authors are also thankful to the Director, Institute of Science, Mumbai for providing
spectral analyses.
Table-1: Characterization of synthesized compounds 2, 3(a-f) and 4.
Compd. R Mol. Formula Yield
(%)
m.p.
(°C )
Spectral data
IR (KBr cm-1
)/1H NMR/
13C NMR (ppm) in
DMSO-d6
2 - C6H6N4O2S2 52 62-64 IR (KBr): 2581 (S-H), 1363 (C=N). 1H NMR:
2.7 (t, 4H, CH2), 9.7 (s, 2H, SH).
[Found: C,35.27, H,2.57, N,24.35, S,27.85%.
Required: C,31.30, H,2.61, N,24.35, S,27.83%.]
3a C6H5 C18H14N4O2 68 114-115 IR (KBr): 1621-1432 (Ar), 1377 (C=N), 1H
NMR: 2.9 (t, 4H, CH2),7.4 (m,10H,ArH)
[Found: C,67.27, H,3.87, N,17.35%.
Required: C,67.92, H,4.4, N,17.61%.]
3b p-OCH3-C6H5 C20H18N4O4 71 97-99 IR (KBr): 1373 (C=N), 1142 (-OCH3). 1H NMR: 2.9 (t, 4H, CH2),3.7 (s, 6H, OCH3), 6.9
(d, 4H), 7.5 (d,4H).13
C NMR: 55.3 (OCH3), 72.4
(CH2),114.3-128.5 (Ar-C), 152.3 (C=N)
[Found: C,63.29, H,4.38, N,14.42%.
Required: C,63.49, H,4.76, N,14.81%.]
3c p-Cl-C6H5 C18H12N4O2Cl2 56 90-94 IR (KBr): 1353 (C=N), 781 (C-Cl).
[Found: C,55.31, H,3.04, N,14.21%.
Required: C,55.81, H,3.1, N,14.47%].
3d o-OH-C6H5 C18H14N4O4 61 124-127 IR (KBr): 3302 (-OH), 1308 (C=N).
[Found: C,61.65, H,4.06, N,16.07%.
Required: C,61.71, H,4.00, N,16.00%].
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Vol.2, No.1, 1-4 (2011)
Vijay V. Dabholkar and Nitin V. Bhusari 3
3e CH=CH-C6H5 C22H18N4O2 58 112-114 IR (KBr): 3021 (CH=CH), 1328 (C=N).
[Found: C,70.99, H,4.53, N,15.01%.
Required: C,71.35, H,4.86, N,15.14%.]
3f p-OCH3-m-OH-
C6H5
C20H20N4O6 57 93-95 IR (KBr): 3311 (-OH), 1331 (C=N),
[Found: C,58.02, H,4.77, N,13.52%.
Required: C,58.25, H,4.85, N,13.59%.].
4 - C18H16N6O2 64 86-89 IR (KBr): 1381 (C=N), 1267 (NH2). 1H NMR: 2.9 (t, 4H, CH2), 8.9 (s, 4H, NH2).
[Found: C,62.10 H,4.48, N,24.06%.
Required: C,62.06, H,4.59, N,24.14%].
Table-2: Antibacterial Activity of compound 2, 3(a-f) and 4
Zone of inhibition (in mm)
Gram Positive Gram negative Comp.
S.aureus C.diphtheria P.aeruginosa E.coli
2 22 20 21 19
3a 21 18 20 18
3b 18 19 18 14
3c 16 18 17 18
3d 21 22 16 17
3e 20 21 18 15
3f 18 19 21 14
4 17 21 21 16
Amphicilin
trihydrate 26 28 24 21
DMSO 0 0 0 0
* Diameter of the disc was 6mm, concentration of the compounds taken was about 100 µg/mL.
NH2
O
N N
O
NN
NH2
O
N N
O
NN
R
R
H2N
HN
O
O
NH
NH2
O
O
O
OC2H5
C2H5
SH
O
N N
O
NN
SH
alc.KOHCAS/MDC
NaHCO3/C2H5OH
(1)
CS2
Ar-CHOCNBr
NH2.NH2.H2OC2H5OH
(2) (3)(4)
Scheme-1
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Vol.2, No.1, 1-4 (2011)
Vijay V. Dabholkar and Nitin V. Bhusari 4
REFERENCES 1 Omar F. A., Mahfouz M.N., Rahman A.M., Eur. J Med. Chem., 31 (1996) 819
2 Franski R., Asian J. Chem., 17 (2005)2063.
3 Narayana B., Vijayraj K. K., Ashalatha B. V., Kumari N. S., Arch. Pharm. (Weinheim) ,338 (2005) 373.
4 Amir M., Kumar S., Acta Pharm., 31 (2007) 57
5 Gaonkar L. S., Rai M. K., Eur. J. Med. Chem., 41 (2006)841.
6 Mishra P., Rajak H., Mehta A., J. Gen. Appl. Microbiol., 51 (2005) 133.
7 Ali M. A.,Yar M. S., Bioorg. Med. Chem. Lett., 17 (2007) 3314.
8 Zargahi A., Tabalabai S. A., Faizi M., Ahadian A., Navabi P., Zanganeh V., Shafiee A., Bioorg. Med.
Chem. Lett., 15 (2005) 1863.
9 Bezerra N. M., De-Oliveira S. P., Srivastava R. M., Da Silva, J. R. Farmaco., 60 (2005) 955.
10 Zareef M., Iqbal R., De Dominquez N. G., Rodrigues J., Zaidi J. H., Arfan M., Supuran C. T., J. Enzyme
Inhib. Med. Chem., 22 (2007)301.
11 Tan T. M., Chen Y., Kang K. H., Bai Li Y., Lim S. G., Ang T. H., Lam Y., Antiviral Res., 71, (2006)7.
12 Hui P. X., Chu H. C., Zhang Y. Z., Wang Q. and Zhang Q., Indian J. Chem., 41B (2002), 2176.
13 Khanum A. S., Shashikanth S., Sudha S. B., Deepak A. S. and Shetty S. H., Pest Manag. Sci., 60 (2004),
1119.
14 Palaska E., Sahin G., Kelicen P., Durlu T. N. and Altinok G., J.R. Farmaco., 57 (2002) 101.
15 Perez S., Lasheral B., Oset C., Carmen A., J. Heterocycl. Chem., 34 (1997) 1527.
16 Hutt M. P., Elslanger E. F., Werbet M. L., J. Heterocycl. Chem., 8 (1970) 511.
17 Baltazzi E., Wysocki A., J. Chem. Ind. , 31 (1963) 1080.
18 Chiba T., Mitsuhiro O., J Org. Chem., 57 (1992) 1375.
19 Shah V. R., Vadodaria M., Parikh A. R., Ind. J Chem., 36B (1997) 101.
20 Bentiss F. and Laqrenee M., J. Heteocycl. Chem., 36 (1999), 1029.
21 Minoo D., Peyman S., Mostafa B. and Mahboobeh B., Tetrahedron Lett., 47 (2006), 6983.
22 Kalluraya B., Jyothi N. Rao and Sujith V. K., Indian J Heterocycl. Chem. ,17 (2008), 359.
23 Cruickshank R., Duguid J. P. and Marmion B. P., Medicinal Microbiology,12th edn, Vol 11, (1975),
(Churchill Livingstone, London).
24 Arthington- Skaggs B. A., Motley M. and Morrison C. J., J. Clin. Microbiology, 38, (2000) 2254.
[IJCEPR-139/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 5-11
January-April, 2011
M. Aravinthraj et al.
Molecular Interaction Studies between H-Bonded Ternary Mixtures of
p-Cresol with simple Aldehydes in Cyclohexane at Different Temperatures
M. Aravinthraj1,*, S. Venkatesan
2 and M. Kamaraj
1
1Department of Physics, Sacred Heart College, Tirupattur, Tamilnadu, India. 2Department of Chemistry, Sacred Heart College, Tirupattur, Tamilnadu, India.
*E-mail: [email protected] Article History:
Received:12 February 2011
Accepted:27 February 2011
ABSTRACT
The ultrasonic velocity (U), Density (ρ) and Viscosity (η) have been measured for three ternary liquid mixtures of P-Cresol +
Cyclohexane + Formaldehyde, P-Cresol + Cyclohexane + Acetaldehyde and P-Cresol + Cyclohexane + Benzaldehyde at 303K,
313K and 323K. The experimental data have been used to calculate the acoustical parameters such as adiabatic
compressibility (β), free length (Lf), free volume (Vf), internal pressure (πi), viscous relaxation time (τ) and Gibb’s free energy
(∆G) were evaluated. The obtained results support the occurrence of dipole-dipole interactions and molecular association through
intermolecular hydrogen bonding in these ternary liquid mixtures.
Key words: Ternary liquid mixture, Acoustical parameter, Molecular association, dipole-dipole interactions. ©2011 ijCEPr. All rights reserved
INTRODUCTION
The studies of multi-component (Binary and ternary liquid) mixtures and solutions have found wide applications in
chemical, textile, leather and nuclear industries [1-3]. The study and understanding of thermo dynamical and
transport properties of liquid mixtures and solutions are more essential for their application in these industries. It
increases interest among several workers for the study of molecular interaction in binary [4, 5] and ternary [6, 7]
liquid mixtures in recent past employing ultrasonic velocity measurement. This precisely helps to understanding the
molecular interactions and structural behavior of molecules and their mixture.
In this paper, we report on the ultrasonic study of four ternary liquid mixtures (p-Cresol + Cyclohexane +
Formaldehyde, p-Cresol + Cyclohexane + Acetaldehyde and p-Cresol + Cyclohexane + Benzaldehyde). The
cyclohexane is a non-polar unassociated, inert hydrocarbons possesses globular structure [8], it has ring structure as
benzene without any S electron and serves as reference point for comparison of the molecular interactions. Cresols
are organic compounds which are methyl phenols. It has isomeric and methyl group substituted onto the benzene
ring of a phenol molecule. It has three forms (ortho, meta and para), one of the o-,m- or p- positions relative to the
OH group. The compounds are highly flammable moderately soluble in water and soluble in ethanol, ether, acetone
cyclohexane and alkalies. Chemically these alkyl phenols undergo electro-philic substitution reactions at the
condensation reaction with aldehydes, ketones [9]. The interactions between cresols and aldehydes may probably be
dipole-dipole or may be due to intra molecular hydrogen bonding [10].
EXPERIMENTAL
The chemicals used in the present work were Analytical reagent (AR) grades with a minimum assay of 99.9%,
obtained from E-Merck (Germany) and Loba chemicals; they are used without further purification. In all the
systems, the various concentrations of the ternary liquid mixtures were prepared in terms of mole fraction, out of
which the mole fraction of the second compound, cyclohexane (X2=0.3) was kept fixed while the mole fraction of
the remaining two (X1 and X3) were varied from 0.0 to 0.7. The ultrasonic velocity was measured by a single crystal
interferometer with a high degree of accuracy operating frequency of 2MHz supplied by M/s. Mittal Enterprises,
New Delhi. Water was circulated around the double walled sample holder to maintain the experimental
temperatures. (Say 303K, 313K and 323K) The density of all compounds was measured by a 10 ml specific gravity
bottle calibrated with double distilled water and acetone. An Ostwald’s viscometer with 10ml capacity was used for
the viscosity measurements of all the compounds. The viscometer was immersed in fresh conductivity water bath
that can be operated at desired temperatures. The flow time of water (tw) and the flow time of solution (ts) were
measured with a digital stop clock with an accuracy of 0.01s (RACER HS-10w).
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Vol.2, No.1, 5-11 (2011)
M. Aravinthraj et al.
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Theory and Calculations
The longitudinal ultrasonic velocity (U), density (ρ) and viscosity (η) of unknown liquid mixtures at any
experimental temperatures are calculated using the following relations. And the density, viscosity of water at
different temperatures was taken from literature [11, 12]. The accuracy in the measurement of density in this method
depends on the accuracy of the weight. The accuracy in the measurement of density is of the order of ± 0.1 kgm-3
.
And the accuracy in the measurement of viscosity depends on the accuracy in the determination of time and density.
The overall accuracy of the measurement of viscosity in this method is ± 0.001Nsm-1
.
Ultrasonic velocity (U)
(1)
Where is the frequency of Ultrasonic wave in Hz, and λ is the wavelength of the Ultrasonic wave in solution
under study in meter.
Viscosity (ηs)
(2)
Where is the co-efficient of viscosity of water in Nsm-1
, is the density of water in Kg/m3, is the flow of
time for water in seconds, is the density of solution in Kg/m3 , and is the flow of time solution in seconds.
Density (ρ2)
(3)
Where are weights of distilled water and experimental liquid and are the densities of water and
experimental liquid.
Using experimentally determined values of ultrasonic velocity (U), density (ρ) and viscosity (η), the following
acoustic and thermodynamic parameters are evaluated.
Adiabatic compressibility (β)
(4)
Where U is the velocity measured in meter/second and ρ is density measured in Kg/m3.
Free Length (Lf)
(5)
Where KT is Jacob’s constant and β is the adiabatic compressibility of a liquid mixtures measured in N-1
m2.
Free Volume (Vf) Suranarayana et al [13] obtained a formula for free volume in term of the ultrasonic velocity (V) and the viscosity of
the liquid (η) as,
(6)
Where Meff is the effective molecular weight ( in which mi and xi are the molecular weight and the
mole fraction of the individual constituents respectively) and K is a temperature independent constant equal to
4.28×109 for all liquids.
Internal Pressure (πi)
On the basis of statistical thermodynamics, Surayanarayana [14] derived an expression for the determination of
internal pressure through the use of the concept of free volume
(7)
Where T is absolute temperature in Kelvin, R is the universal gas constant and b is the cubic packing fraction factor
is assumed to be ‘2’ for all liquid systems.
Viscous relaxation time (ττττ)
(8)
Where η is the viscosity of the solution in Nsm-2
and β is adiabatic compressibility in N-1
m2
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Vol.2, No.1, 5-11 (2011)
M. Aravinthraj et al.
7
Gibb’s free energy (∆G)
The Gibb’s free energy of activation flow in the mixtures can be obtained on the basis of Erying rate process theory
and it can be able to calculate from the relation,
(9)
Where τ is the viscous relaxation time measured in meters, K is the Boltzmann’s constant, h is a Plank’s constant
and T is the temperature measured in kelvin.
RESULT AND DISCUSSION The experimentally measured values of density, viscosity and ultrasonic velocity of the three ternary systems of p-
Cresol + Cyclohexane + Formaldehyde, p-Cresol + Cyclohexane + Acetaldehyde and p-Cresol + Cyclohexane +
Benzaldehyde at 303K, 313K and 323K are presented in Table-1. The systems taken for the present study is
basically an H-Bonded complexes of phenol (p-cresol) with aldehydes in cyclohexane. Here mole fraction of
cyclohexane is fixed as 0.3 and the mole fraction of the other two components are varied from 0.0 to 0.7.
Acoustical parameters such as adiabatic compressibility (β), intermolecular free length (Lf), free volume (Vf),
internal pressure (πi), kinetic parameter like viscous relaxation time (τ) and thermo dynamical parameter like Gibb’s
Free Energy (∆G) were calculated from the measured ultrasonic velocity, density and viscosity using standard
equations given in theory and calculations.
From the measured data, it is observed that there is a sudden fall of density in all the systems for second molar
concentration, and from the second concentration onwards p-cresol is made to dissolve in cyclohexane for which
aldehydes are used as doped. When aldehyde and cyclohexane are taken without p-cresol, the density is found to be
higher. This serves the possibility of greater interaction between the aldehydes and solvent due to dipole-dipole
interaction as well as possibility of weak H-bond shown in Figure (a).
The solvent has hydrogens placed at the 1,3-diaxial position, such that any of these hydrogens are free to interact
with non-bonding electrons (or) n-electron from the carbonyl oxygen of aldehydes Figure (b). This serves to the
increasing density with the increasing of aldehydes concentration. From the second molar concentration onwards a
gradual increasing trend in density is observed. Figure (a) shows the interaction between Oxygen of the p-cresol and
flagpole Hydrogen of the Cyclohexane. This dipole-dipole interaction serves for the gradual increase in density.
Fig. (a) Fig. (b)
From the second concentration onwards the influence of non bonding electron is frankly reduced when p-cresol is
began to introduce. Another kind of factor is observed when the mole fraction of aldehydes is taken as 0(zero), and
the mole fraction of p-cresol is taken as 0.7, here also the density is greater because of the possible interaction
between the non bonding electrons of oxygen and the cyclohexane. The mole fraction of cyclohexane is taken as 0.3
M, which is constant for the overall experiment. From the second concentration onwards p-cresol and aldehydes are
mixed with different molar concentration in cyclohexane, and then increasing trend is observed in density, viscosity
and velocity. This is because of more hydrogen bonding and dipole-dipole interaction observed as shown in Fig (c)
and Fig (d) respectively. This behavior at such concentrations is same as the ideal mixtures behavior can be
attributed to intermolecular interactions in the systems studied [16]. It is evident from table-1.
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Vol.2, No.1, 5-11 (2011)
M. Aravinthraj et al.
8
Fig. (c) Fig. (d)
It is found that from the table-2, the adiabatic compressibility and intermolecular free length decreases with increase
in concentration of p-cresol in all the three systems. This shows an inverse behavior as compared to ultrasonic
velocity. This indicates that there is a significant interaction between solute and solvent molecules. It can be taken as
the indication of complex formation.
The addition of interacting molecules breaks up the molecular clustering of the other, releasing several interaction,
because p-cresol has the possibility of forming intermolecular H-bonding with such carbonyl oxygen. However free
volume increases with increase in molar concentration of p-cresol, the p-cresol system has the lone pair of electron
in oxygen of p-cresol system interact with Hydrogen in hydroxyl group of another p-cresol system. This is clearly
observed in the increasing trend in free volume with increasing molar concentration of p-cresol. At the same time
the mole fraction of doped is purposely decreased, because this factor holds well only from the introduction of p-
cresol.
From Table-3 it is found that, internal pressure decreases with increase in concentration of p-cresol, because the
hydrogen releasing ability of p-cresol is becoming low. The decreasing density with the increase in concentration of
p-cresol and the decreasing concentration of aldehydes is contributed from the above explanation. Also a decrease
in internal pressure with the decrease in size of the alkyl group of aldehyde is observed. This is naturally due to the
increasing available space with the decreasing size of alkyl group.
However, the viscous relaxation time increases with increasing molar concentration of p-cresol as shown in Table -3
and a decrease in all these values is also noted with the increasing temperature. This is clearly due to the increasing
free space between the molecules and the weakening of intermolecular forces. Generally, viscous relaxation time is
of the order of 10-12
s, for identifying structural relaxation between the component molecules. This shows the
presence of molecular interactions [17], Gibb’s free energy confirms the same, which indicates the need for smaller
time for the cooperative process or the rearrangement of the molecules in the mixtures.
Table -1: Density (ρ), Viscosity (η) and Velocity (U) of p-Cresol + Cyclohexane + Formaldehyde, p-Cresol +
Cyclohexane + Acetaldehyde and p-Cresol + Cyclohexane + Benzaldehyde at 303K, 313K and 323K
Density (ρ) Viscosity (η) Velocity (U) Mole fraction
(Kg/m3) (10
-3Nsm
-2) (ms
-1)
X1 X2 303K 313K 323K 303K 313K 323K 303K 313K 323K
p-Cresol + Cyclohexane + Formaldehyde
0.000 0.705 887.7 883.8 880.3 1.704 1.339 1.106 1310.1 1306.5 1298.1
0.100 0.606 846.2 841.1 837.0 2.890 2.344 1.829 1323.9 1318.7 1309.9
0.197 0.498 870.6 866.1 861.2 2.955 2.389 1.853 1338.1 1330.0 1318.9
0.301 0.403 871.3 867.9 863.3 3.112 2.398 1.899 1345.9 1339.2 1323.2
0.399 0.297 873.0 867.9 864.0 3.075 2.435 1.911 1357.3 1348.9 1336.8
0.497 0.195 875.1 870.3 866.8 3.232 2.458 2.023 1364.5 1353.1 1349.3
0.607 0.104 877.0 873.5 869.8 3.345 2.701 1.985 1374.7 1363.4 1354.8
0.705 0.000 879.0 875.2 870.6 3.538 2.901 2.189 1361.0 1357.3 1349.3
p-Cresol + Cyclohexane + Acetaldehyde
0.000 0.701 871.2 868.1 863.9 1.415 1.050 0.880 1309.3 1300.5 1295.1
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Vol.2, No.1, 5-11 (2011)
M. Aravinthraj et al.
9
0.102 0.604 842.1 837.5 830.3 2.190 1.845 1.407 1319.8 1310.1 1301.9
0.199 0.501 867.1 852.8 847.7 2.503 2.106 1.524 1326.9 1320.1 1312.1
0.305 0.398 868.3 863.3 859.5 2.621 2.161 1.577 1333.8 1328.6 1322.4
0.398 0.301 870.3 866.8 862.3 2.859 2.223 1.590 1349.7 1337.5 1331.6
0.504 0.200 874.4 869.3 865.8 3.105 2.236 1.777 1356.4 1350.0 1339.6
0.601 0.098 875.0 872.5 867.2 3.256 2.388 1.954 1360.0 1354.0 1347.8
0.700 0.000 879.0 875.5 869.6 3.506 2.896 2.185 1360.1 1355.5 1350.9
p-Cresol + Cyclohexane + Benzaldehyde
0.000 0.700 870.2 868.5 867.3 1.024 0.885 0.773 1307.9 1304.5 1300.3
0.101 0.603 838.2 835.2 832.3 1.216 1.004 0.890 1310.7 1296.0 1289.1
0.203 0.501 858.8 855.2 851.8 1.426 1.193 1.046 1321.7 1305.8 1295.0
0.299 0.404 860.1 859.8 857.4 1.711 1.362 1.199 1329.7 1310.7 1302.2
0.403 0.308 869.3 867.2 865.9 1.950 1.604 1.315 1338.4 1318.8 1309.4
0.503 0.199 870.0 868.9 864.6 2.427 2.025 1.560 1345.2 1331.9 1320.5
0.599 0.104 874.4 872.9 871.8 2.879 2.463 1.758 1350.4 1333.6 1326.0
0.703 0.000 878.0 876.9 872.0 4.096 2.850 2.223 1359.5 1351.4 1328.6
Table -2: Adiabatic compressibility (β), Free Volume (Vf), and Free Length (Lf) of p-Cresol + Cyclohexane +
Formaldehyde, p-Cresol + Cyclohexane + Acetaldehyde and p-Cresol + Cyclohexane + Benzaldehyde at 303K,
313K and 323K
Adiabatic compressibility
(β)
Free length
(Lf)
Free Volume
(Vf) Mole fraction
(10-10
N-1
m2) (10
-10m) (10
-7 m
3 mol
-1)
X1 X2 303K 313K 323K 303K 313K 323K 303K 313K 323K
p-Cresol + Cyclohexane + Formaldehyde
0.000 0.705 6.5630 6.6280 6.7406 5.0564 5.0814 5.1243 0.1719 0.2457 0.3241
0.100 0.606 6.7417 6.8365 6.9626 5.1248 5.1607 5.2080 0.0908 0.1236 0.1776
0.197 0.498 6.4143 6.5264 6.6750 4.9988 5.0423 5.0993 0.1045 0.1424 0.2059
0.301 0.403 6.3355 6.4244 6.6158 4.9680 5.0027 5.0767 0.1158 0.1700 0.2368
0.399 0.297 6.2174 6.3316 6.4759 4.9215 4.9665 5.0227 0.1461 0.2054 0.2915
0.497 0.195 6.1368 6.2751 6.3367 4.8895 4.9443 4.9684 0.1719 0.2561 0.3414
0.607 0.104 6.0332 6.1580 6.2630 4.8480 4.8979 4.9395 0.2121 0.2886 0.4538
0.705 0.000 6.1412 6.2017 6.3090 4.8912 4.9153 4.9576 0.2670 0.3584 0.5419
p-Cresol + Cyclohexane + Acetaldehyde
0.000 0.701 6.6950 6.8109 6.9006 5.1070 5.1510 5.1848 0.3690 0.5713 0.7399
0.102 0.604 6.8164 6.9559 7.1049 5.1531 5.2055 5.2610 0.2160 0.2763 0.4108
0.199 0.501 6.5500 6.7283 6.8512 5.0514 5.1197 5.1662 0.1998 0.2569 0.4137
0.305 0.398 6.4735 6.5618 6.6524 5.0218 5.0559 5.0907 0.2143 0.2846 0.4533
0.398 0.301 6.3074 6.4486 6.5395 4.9570 5.0121 5.0473 0.2182 0.3139 0.5154
0.504 0.200 6.2175 6.3114 6.4360 4.9215 4.9585 5.0072 0.2265 0.3683 0.5137
0.601 0.098 6.1789 6.2513 6.3472 4.9062 4.9349 4.9726 0.2509 0.3968 0.5324
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Vol.2, No.1, 5-11 (2011)
M. Aravinthraj et al.
10
0.700 0.000 6.1495 6.2163 6.3011 4.8945 4.9210 4.9545 0.2703 0.3583 0.5438
p-Cresol + Cyclohexane + Benzaldehyde
0.000 0.700 6.7172 6.7653 6.8191 5.1155 5.1337 5.1541 1.5849 1.9652 2.3944
0.101 0.603 6.9437 7.1278 7.2292 5.2010 5.2695 5.3068 1.2331 1.6165 1.9208
0.203 0.501 6.6652 6.8571 6.9996 5.0956 5.1685 5.2219 0.9860 1.2654 1.5216
0.299 0.404 6.5750 6.7692 6.8774 5.0610 5.1352 5.1761 0.7586 1.0451 1.2539
0.403 0.308 6.4210 6.6292 6.7353 5.0014 5.0818 5.1223 0.6322 0.8285 1.1041
0.503 0.199 6.3511 6.4868 6.6325 4.9741 5.0270 5.0831 0.4594 0.5938 0.8670
0.599 0.104 6.2709 6.4413 6.5228 4.9426 5.0093 5.0409 0.3587 0.4447 0.7310
0.703 0.000 6.1616 6.2441 5.3932 4.8993 4.9320 4.5837 0.2141 0.3655 0.5946
Table -3: Internal Pressure (πi), Viscous Relaxation Time (τ) and Gibb’s free energy (∆G) of p-Cresol +
Cyclohexane + Formaldehyde, p-Cresol + Cyclohexane + Acetaldehyde and p-Cresol + Cyclohexane +
Benzaldehyde at 303K, 313K and 323K
Internal pressure
(πi)
Viscous Relaxation Time
(τ)
Gibb’s Free
Energy (∆G) Mole fraction
(10-6
Pa) (10-12
s) (10-20
KJ mol-1
)
X1 X2 303K 313K 323K 303K 313K 323K 303K 313K 323K
p-Cresol + Cyclohexane + Formaldehyde
0.000 0.705 14.090 12.472 11.342 1.4911 1.1836 0.9943 0.4077 0.3898 0.3686
0.100 0.606 15.867 14.397 12.878 2.5982 2.1373 1.6985 0.5086 0.4948 0.4722
0.197 0.498 14.769 13.754 11.907 2.5280 2.0793 1.6497 0.5036 0.4897 0.4666
0.301 0.403 12.561 11.314 9.971 2.6295 2.0545 1.6758 0.5017 0.4874 0.4696
0.399 0.297 10.921 9.529 8.429 2.5493 2.0563 1.6505 0.5051 0.4876 0.4667
0.497 0.195 9.357 8.127 6.977 2.6448 2.0565 1.7094 0.5118 0.4857 0.4735
0.607 0.104 7.818 6.712 6.034 2.6908 2.2184 1.6581 0.5149 0.5018 0.4676
0.705 0.000 6.829 5.661 4.915 2.8985 2.3989 1.8416 0.5284 0.5165 0.4879
p-Cresol + Cyclohexane + Acetaldehyde
0.000 0.701 8.691 7.495 6.854 1.2634 0.9539 0.8101 0.3776 0.3435 0.3289
0.102 0.604 9.684 8.887 7.742 1.9907 1.7115 1.3336 0.4602 0.4531 0.4254
0.199 0.501 9.630 8.997 7.603 2.1867 1.8898 1.3923 0.4773 0.4717 0.4337
0.305 0.398 8.883 7.948 6.873 2.2624 1.8907 1.3992 0.4834 0.4718 0.4347
0.398 0.301 8.345 7.268 6.096 2.4048 1.9118 1.3870 0.4945 0.4739 0.4330
0.504 0.200 7.719 6.539 5.837 2.5746 1.8818 1.5252 0.5069 0.4709 0.4514
0.601 0.098 6.924 5.931 5.356 2.6831 1.9910 1.6542 0.5144 0.4815 0.4671
0.700 0.000 6.782 5.664 4.907 2.8753 2.4005 1.8364 0.5269 0.5166 0.4873
p-Cresol + Cyclohexane + Benzaldehyde
0.000 0.700 3.464 3.220 3.012 0.9178 0.7988 0.7035 0.3196 0.3102 0.3016
0.101 0.603 3.669 3.344 3.150 1.1262 0.9543 0.8582 0.3568 0.3436 0.3401
0.203 0.501 4.013 3.682 3.453 1.2676 1.0910 0.9767 0.3782 0.3687 0.3651
0.299 0.404 4.379 3.605 3.362 1.5007 1.2301 1.0996 0.4089 0.3912 0.3881
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Vol.2, No.1, 5-11 (2011)
M. Aravinthraj et al.
11
0.403 0.308 4.679 4.269 3.875 1.6696 1.4183 1.1814 0.4283 0.4179 0.4019
0.503 0.199 5.203 4.773 4.193 2.0560 1.7520 1.3800 0.4661 0.4575 0.4320
0.599 0.104 5.663 5.265 4.458 2.4073 2.1157 1.5297 0.4947 0.4929 0.4519
0.703 0.000 6.737 5.632 4.771 3.3652 2.3731 1.5989 0.5555 0.5145 0.4605
CONCLUSION From the experimental and calculated parameters, there is a significant interactions observed only after introducing
p-Cresol. This increases density, viscosity and velocity of all the liquid mixtures. This increasing trend is due to the
intermolecular hydrogen bonding between carbonyl oxygen of aldehydes with hydroxyl hydrogen in p-cresol and
dipole- dipole interaction between the same compounds with carbonyl carbon of aldehydes and hydroxyl oxygen.
Hence this investigation provides comprehensive idea about the molecular interactions between solute and solvent.
The order of interactions is found to be Formaldehyde > Acetaldehyde > Benzaldehyde.
REFERENCE
1. Rowlinson J.S., and Swinton P. L., 1982. Liquids and liquid mixtures. 3rd
edition, Butterworth scientific,
London.
2. Acree W.E., 1984. Thermodynamic properties of Non-electrolytic solutions, academic press, New York.
3. Prausnitz J.M., Linchenthalr and Azevedo E.G., 1986. Molecular thermodynamics of fluid phase equlibria.
2nd
edition, Engle wood cliffs, Prentic Hall Inc.
4. Sastry N.V., and Patel S.R., Int. J. Thermo phys, 21(5)(2000).
5. Prabakar S., and Rajagopal., Ind J pure & appli ultra, 27, (2005), 4.
6. Sharma V.K., Kalra., Romi and Kotach., Ind J Chem., 42A, 292.
7. Asghar J., Liakath Ali Khan F and Subramani K., Rasaya J Chem, 3(4)( 2010), 697.
8. Ali, A.S. Adida Hydar and A.K Nain., Ind J Phys, 76B (15), (2002), 661.
9. Thirumaran S and Deepesh George., ARPN, 4(4)(2009), 1.
10. Jayakumar S., Karunanithi S and Kannappan V., Ind J pure & Appli Phys, 34(1996) 761.
11. Riddick J.A., Bunger W.B and Sanano T.K., 1984. Techniques in Chemistry, Vol. II, Organic Solvent. 4th
edition, John Willey, New York.
12. Hand book of chemistry and physics, 1984-85, 65th
edition, the chemical rubber company, Cleveland, Ohio,
USA.
13. Surayanarayana. C.V. and kuppusamy.T , J.Acou.soc.Ind, 4, (1976), 75.
14. Surayanarayana. C.V. ,J.Acou.soc.Ind, 7, (1979), 131.
15. Baldev Raj., Rajendran V and Planichami P ., 2004. Science and technology of ultrasonics., 3rd
edition,
Narosa publication, India.
16. Guptha A K, Krishnakumar and Birendra Kumar Karn, J.Ind. Council.Chem, 26(1)(2009), 77.
17. Kannappan A N, Keasvasamy R and Ponnuswamy V, Am.J Eng and Appli.Sci, , 1(2)( 2008), 95.
[IJCEPR-151/2011]
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International Journal of ChemicalChemicalChemicalChemical, Environmental and , Environmental and , Environmental and , Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 12-19
January-April, 2011
N. Kumar et al.
Chemical Analysis on Different Oils Use in Tyre Tread Cap Compound
N. Kumar*, R.K. Khandelwal1, P.L. Meena
1, K. S. Meena
1, T.K. Chaki
2 , D.K. Mahla
2
and S. Dasgupta3
1P.G.Department of Chemistry, M.L.V.Govt.P.G.College, Bhilwara-311001 Rajasthan 2Indian Institute of Technology Kharagpur West Bengal-721302, India 3J K Tyre HASETRI Kankroli, Rajasthan
*E-mail: [email protected] Article History:
Received: 17 March 2011
Accepted:23 March 2011
ABSTRACT
The global market place is increasingly demanding safe process oils to reduce the environmental impact of tires. The replacement
of classified distillate aromatic extracts by non-carcinogenic MES, TDAE, or naphthenic process oils will reduce the PAH
emissions. . In the present work three types of low PCA and one regular high PCA Petroleum oils were chemically analyzed. The
oils were characterized for different chemical analysis . These low PCA oils can act as the best alternative processing aids for
rubber industry. The rheological, properties of SSBR loaded with different LPCA & HPCA oils have been studied. in order to
obtain similar properties. The data show that the best results are obtained using LPCA. A comparative study has been carried
out on SSBR filled with various oils.
Key words: low PCA oils, Polycyclic aromatics, carcinogenesis, PAH, risk assessment. ©2011 ijCEPr. All rights reserved
INTRODUCTION In compounding rubber and rubber composition for use in pneumatic tyre, it is common to utilize processing oils to
soften and extend the rubber[7]. Typical, aromatic processing oils, having a certain content of poly aromatic
compound or polyaromatic hydrocarbons ,have bee used .recently , regulatory concerns have necessisted the use of
processing oils having alower PCA content.
Rubber formulations used in various tyre components previously have been designed using conventional processing
oils .How ever, in changing to the use of the lower PCA content oils ,some lose in rubber compound performance is
noted. It is , there for necessary to develop new rubber compounds that provide desirable performance levels wile in
corporating the use low PCA oils[8].
EXPERIMENTAL Materials studied are given in Table 1.
Physicochemical characterization
The oils were characterized for Specific gravity by hydrometer (ASTM D 1298) , flash and fire point (ASTM D92),
pour point (ASTM D97), specific gravity (ASTM D1298), saybolt viscosity (ASTM D88), Fourier transform
infrared (FTIR) spectroscopic study of the petroleum oils was performed in a FTIR System from PERKIN ELMER,
USA for checking functional groups present[1-3].
Compound mixing Mixing of rubber compound was carried out using a two-wing rotor laboratory Banbury mixer (Stewart Bolling,
USA) in three stages (master batch remill and final batch) and the formulations are given in Table
Master batch mixing was done setting the temperature control unit (TCU) at 90°C and rotor speed at 60 rpm.. After
the power integrator (PI) indicated achievement of 0.32 kWh, the master batch was dumped. The dump temperature
of the master batches was found to be within 140 - 150°C. The master batches were sheeted out in a laboratory two-
roll mill. Further mixing of the master batches were carried out after a maturing period of 8 hours[8].
For final batch mixing, the TCU was kept at 600C and rotor speed at 30 rpm. The earlier prepared master batch was
mixed with sulfur, accelerator and scorch inhibitor. The batch was dumped at a PI reading of 0.12 kWh. The dump
temperature of the batches was found to be within 95 – 105°C. The final batches were also sheeted out on a
laboratory two-roll mill8-12
Formulation is according to Table 2.
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Table-1: Material Required
1 S SBR having regular aromatic oil
2 S SBR having low PCA oil (3830)
3 Regular oil and LPCA OIL No.1,2,3,
4 Filler N339 black
5 ZnO
6 Stearic Acid
7 6PPD
8 MC Wax
9 MS 40
10 S
11 TBBS
12 DCBS
13 PVI
RESULTS AND DISCUSSION 1. Flash and Fire Point
The flash point of a volatile liquid is the lowest temperature at which it can vaporize to form an ignitable mixture in
air. Measuring a liquid's flash point requires an ignition source. At the flash point, the vapor may cease to burn when
the source of ignition is removed. The flash point is not to be confused with the autoignition temperature, which
does not require an ignition source. The fire point, a higher temperature, is defined as the temperature at which the
vapor continues to burn after being ignited. Neither the flash point nor the fire point is related to the temperature of
the ignition source or of the burning liquid, which are much higher.
The flash point is often used as a descriptive characteristic of liquid fuel, and it is also used to help characterize the
fire hazards of liquids. “Flash point” refers to both flammable liquids and combustible liquids. There are various
standards for defining each term. The flash and fire point results are shown in Table 3.
Flash and fire point is one of the important criteria for determining the process safety while handling the rubber
compound during mixing, calendaring, extrusion etc. Higher flash and fire point of oils always indicates good
process safety. High flash and fire point of oils may be due to presence of carbonyl groups, alkaloids groups
etc[11].
2. Pour Point
The pour point of a liquid is the lowest temperature at which it will pour or flow under prescribed conditions. It is a
rough indication of the lowest temperature at which oil is readily pumpable.
Also, the pour point can be defined as the minimum temperature of a liquid, particularly a lubricant, after which, on
decreasing the temperature, the liquid ceases to flow.The pour point results are shown in Table 4. All the Low PCA
oils show pour point less than 0°C except aromatic oil. Lower pour point improves the handling of oils during
winter season. Surrounding temperature during winter season reduces drastically when some processing oils require
heating arrangement for ease of flow to the Banbury chamber for mixing. Additional energy consumption is required
for such heating process. With natural oils, such additional processing costs can be eliminated.
3. Saybolt viscosity
Viscosity describes a fluid's internal resistance to flow and may be thought of as a measure of fluid friction. For
example, high-viscosity felsic magma will create a tall, steep stratovolcano, because it cannot flow far before it
cools, while low-viscosity mafic lava will create a wide, shallow-sloped shield volcano. All real fluids (except
superfluids) have some resistance to stress and therefore are viscous, but a fluid which has no resistance to shear
stress is known as an ideal fluid or inviscid fluid.
The study of flowing matter is known as rheology, which includes viscosity and related concepts.The saybolt
viscosity results are shown in Table 5. High Saybolt viscosity indicates higher aromaticity.
4. Aniline Point
Aniline point is defined as the temperature at which equal volumes of aniline(C6H5NH2) and diesel oil are
completely miscible. The value gives an indication of the aromatic content of diesel oil, since aniline is an aromatic
compound which is dissolved on heating by the aromatics in diesel oil. The greater the aniline point, the lower the
aromatics in diesel oil. A higher aniline point also indicates a higher proportion of paraffin. The diesel index is
directly related to aniline point as:
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N. Kumar et al.
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DIESEL INDEX = ((ANILINE POINT(DEG F))(API GRAVITY))/100
A higher aniline point (and therefore a lower aromatic content) in diesel oil is desirable, in order to prevent
autoignition in diesel engines.In cases where the Aromatic content in the oil is very high, in such cases "Mixed
Aniline Point" needs to me measured to determine the approximate content of Aromatic in the oil.The aniline point
results are shown in Table 6.
All the Low PCA oils show higher values whereas High PCA oils show lower values. Aniline point indicates the
presence of aromatic ring in the oils. Higher the aromatic groups lower the aniline point.
5. Specific gravity
Specific gravity is the heaviness of a substance compared to that of water, and it is expressed without units. In the
metric system specific gravity is the same as in the English system[10].
In relationship to liquids, the term specific gravity is used to describe the weight or density of a liquid compared to
an equal volume of fresh water at 4°C (39° F). If the liquid you are comparing will float on this water it has a
specific gravity of less than one. If it sinks into the fresh water the specific gravity is more than one. As you have
already guessed fresh water at 4°C (39° F) has been assigned a value of one. The specific gravity results are shown
in Table 7.
6. Aromatic content (CA)
The aromatic content results are shown in Table 8.Higher aromatic content is basically the presence of
polycyclic group in the oils.
7. FTIR Study for surface group
IR bands of aromatic C-H stretching at 3010 and 3080 cm-1
and overtone and combination band due to C-H out-of-
plane at 1600-2000 cm-1
were observed. Also, IR band of aromatic ring CC stretching and aromatic C-H in-plane
appeared in 1000-1200 cm-1
region[9].The FTIR results are shown in Table 9.
CONCLUSION The recent change in world scenario in shifting towards low PCA oils and restriction on high PCA oils. Present
study is focused on chemical, analytical and compound characterization of petroleum oils. These oils were found to
be suitable on the basis of low PCA content.. All non-carcinogenic process oils contain very low concentrations of
polycyclic aromatic hydrocarbons and meet the 1 mg/kg benzo[a]pyrene limit set by the VDA. Hence, the
replacement of HPCA by non-carcinogenic process oils in oil extended natural or synthetic rubber and therefore in
finished tires will reduce the PAH emission from tire wear by more than 98 %. Test results are intended to support
the rubber and tire industries in their environmental challenge to replace the classified aromatic oils. Further
extensive compounding and evaluation work will be required by each company using its proprietary tire formulation
technology. Demand for these oils is expected to rise as car manufacturers realise that carcinogenic emissions from
tires can hereby be greatly reduced. It has demonstrated on a commercial scale that this challenge can be met by a
change to safer alternatives such as LPCA . The production LPCA oil are already on the market[12].
ACKNOWLEDGEMENTS The author would like to thank IIT Kharagpur (W.B.) & J K Tyre HASETRI Kankroli, Rajasthan for excellent
cooperation, extensive evaluations and discussion.
210
220
230
240
250
260
270
Aro.oil
Reg.
LP
CA-1
LP
CA-2
LP
CA-3
Flash and fire point (°C)
Flash and fire point
(°C)
Fig.-1: Flash and fire point
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Table-2: Formulation
Table-3: : Flash and fire point
Name of oils Flash and fire point(°C)
Aromatic oil regular 235
Low PCA oil No.1 240
Low PCA oil No.2 230
Low PCA oil No.3 267
Table-4: Pour point
Name of oils Pour point (°C)
Aromatic oil regular 7
Low PCA oil No.1 -8
Low PCA oil No.2 4
Low PCA oil No.3 -4
Table-5: Saybolt viscosity
Name of oils Saybolt viscosity(sec)
Aromatic oil regular 130
Low PCA oil No.1 105
Low PCA oil No.2 65
Low PCA oil No.3 120
Ingredients HPCA LPCA-1 LPCA-2 LPCA-3
RMA4 27 27 27 27
BR 35 35 35 35
VSL5525 52 -- -- --
Tufden3830 -- 52 52 52
N339 60 60 60 60
Reg Ar. Oil 5 -- -- --
LPCA-1 -- 5 -- --
LPCA-2 -- -- 5 --
LPCA-3 -- -- -- 5
ZnO(WS) 2.25 2.25 2.25 2.25
St Acid 0.5 0.5 0.5 0.5
6PPD 1.9 1.9 1.9 1.9
MC Wax 2.4 2.4 2.4 2.4
MS 40 1 1 1 1
S(108) 2.2 2.2 2.2 2.2
TBBS 1.2 1.2 1.2 1.2
DCBS 0.6 0.6 0.6 0.6
PVI 0.15 0.15 0.15 0.15
Batch weight 191.2 191.2 191.2 191.2
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Table-6: Aniline point
Name of oils Aniline point (°C)
Aromatic oil regular 46.5
Low PCA oil No.1 100
Low PCA oil No.2 30
Low PCA oil No.3 105
Table -7: Specific gravity
Name of oils Specific gravity
Aromatic oil regular 1.001
Low PCA oil No.1 0.916
Low PCA oil No.2 0.938
Low PCA oil No.3 0.921
Table -8: Aromatic content
S.No. Carbon type
analysis (%)
Low
PCA
1
Low
PCA
2
Low
PCA 3
Aromatic
oil
1
2
3
CA
CP
C N
19.8
59.8
20.4
-
-
-
16.1
68.4
15.5
36.8
58.7
4.5
Table-9: FTIR study
Name of oils Surface group present
Aromatic oil
regular
Alkyl group –CH2-R St.
Aromatic substituent C-H St.
Low PCA oil
No.1
Alkyl group CH2-R St.
Alkyl group CH2-R St.
Aromatic substituent C-H St.
Low PCA oil
No.2
Aliphatic hydrocarbon C-H St.
Aliphatic hydrocarbon –CH2-R St.
Aromatic substituent C-H St
Low PCA oil
No.3
Aliphatic hydrocarbon St.
Aliphatic hydrocarbon (Short chain compound or substituent)
210
220
230
240
250
260
270
Aro.oil
Reg.
LP
CA-1
LP
CA-2
LP
CA-3
Flash and fire point (°C)
Flash and fire point
(°C)
Fig.-2: Flash and fire point
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-8
-6
-4
-2
0
2
4
6
8
Aro.oil
Reg.
LPCA-
1
LPCA-
2
LPCA-
3
Pour point (°C)
Pour point
(°C)
Fig.-3: Pour point
0
20
40
60
80
100
120
140
Aro.oil
Reg.
LPCA-
1
LPCA-
2
LPCA-
3
Saybolt viscosity (sec)
Saybolt viscosity
(sec)
Fig.-4: Saybolt viscosity
0
20
40
60
80
100
120
Aro.oil
Reg.
LPCA-
1
LPCA-
2
LPCA-
3
Aniline point (°C)
Aniline point (°C)
Fig.-5: Aniline point
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0.86
0.88
0.9
0.92
0.94
0.96
0.98
1
1.02
Aro.oil
Reg.
LPCA-
1
LPCA-
2
LPCA-
3
Specific gravity
Specific gravity
Fig.-6: The specific gravity
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
0.0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
86.1
cm-1
%T
2906.97
2729.56
2360.48
1907.41
1601.68
1460.29
1377.05
1166.60
1060.17
1032.83
960.56
864.49
810.24
749.81
728.31
577.79
428.26
3482.62
2727.09
2670.21 1605.36
1460.29
1376.76
1306.43
1168.61
1031.86
965.14
869.22
814.48
722.55
564.38
2870.61
2728.51
2360.15
1603.27
1460.26
1376.91
1306.59
1156.33
1031.85
963.58
867.54
813.52
726.71
576.98
463.88
Fig.-7: FTIR of oils: Black color high PCA/ Blue color Oil No1/ Red color Oil 2
Table-10: Rheometric Properties @ 1930C/2.5min
TEST HPCA LPCA-1 LPCA-2 LPCA-3
MIN TQ. (lb-in) 0.23 0.23 0.22 0.22
MAX.TQ.(lb-in) 1.19 1.84 1.87 1.82
Final TQIlb-in) 13.61 13.18 13.06 13.26
tS1 (min) 0.49 0.49 0.5 0.48
tS2 (min) 0.66 0.64 0.64 0.63
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Vol.2, No.1, 12-19 (2011)
N. Kumar et al.
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Table-11: Rheometric Properties @ 160 C/30min (Final)
REFERENCES
1. Internet “Definitions of terms relating to oil”.
2. ASTM D1566-06, “Standard Terminology Relating to Rubber”.
3. Encyclopedia of Polymer Science and Engineering, “Cellular Materials to Composites”, IInd
Edition,
A Wiley-Interscience Publication, 3, 619 (1985).
4. ASTM D2230-96 (Reapproved 2002), “Rubber property-Extrudability of Unvulcanised
Compounds”.
5. J. S. Dick and H. Pawlowski, Rubber World, 211(1995) 20.
6. A.Y. Coran and J. B. Donnet, Rubber Chem. Tech., 65(1992) 973.
7. H. D. Luginsland, J. Frohlich and A. Wehmeier, Rubber Chem. Tech.75(2002) 563.
8. S. Das Gupta, presented at 19th
Indian Rubber Manufacturers Research Association (IRMRA) Conference”,
p. 187, Bombay, India, December 2005.
9. S. Das Gupta, S. L. Agrawal and R. Mukhopadhyay, “Proceedings of the 19th
Indian Rubber Manufacturers
Research Association (IRMRA) Conference, Bombay, India, December 2005.
10. en.wikipedia.org/wiki/Oil
11. United State patent No. US6,984,687,B2 Jan.10,2006.
12. J. Fraser Stoddart a,*, Howard M. Colquhoun, Tetrahedron 64 (2008) 8231.
[IJCEPR-153/2011]
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tC10 (min) 0.53 0.51 0.52 0.5
tC40 (min) 0.83 0.8 0.81 0.8
tC50 (min) 0.88 0.84 0.86 0.84
tC90 (min) 1.19 1.15 1.17 1.14
TEST HPCA LPCA-1 LPCA-2 LPCA-3
MIN TQ. (lb-in) 2.45 2.67 2.64 2.76
MAX.TQ.(lb-in) 15.84 15.76 15.4 16.09
Final TQ.(Ilb-in) 14.56 14.21 13.88 14.57
tS1 (min) 4.1 4.09 4 4.22
tS2 (min) 5.07 4.94 4.74 5.08
tC10 (min) 4.59 4.47 4.32 4.64
tC40 (min) 5.69 5.58 5.38 5.77
tC50 (min) 5.87 5.77 5.59 5.97
tC90 (min) 7.79 7.66 7.57 7.85
Max-Min Tq.(lb-in) 13.39 13.09 12.76 13.33
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 20-25
January-April, 2011
Jitendra H. Deshmukh and M. N. Deshpande
Infra Red Spectral and X-ray Diffraction Study of Fe (II), Co(II), Cu (II),
Metal Chelates with N1-(5, 6-dimethoxypyrimidin-4-yl) Sulphanilamide
Jitendra H. Deshmukh*1 and M. N. Deshpande
2
1Department of chemistry, Yeshwant Mahavidyalaya, Nanded-431602 2Head P.G. Department of chemistry, NES, Science College, Nanded-4316053
*E-mail: [email protected] Article History:
Received: 14 April 2011
Accepted:25 April 2011
ABSTRACT N1-(5, 6-dimethoxypyrimidin-4-yl) sulphanilamide (DMPS)forms chelates having general formula [Fe (DMPS)2 Cl2] H2O, [Co
(DMPS)Cl22H2O] and [Cu(DMPS)ClH2O]Cl .Infra red ,electronic and X-ray diffraction study of Fe(II), Co(II), Cu(II) metal
chelates.
Keywords: N1-(5,6-dimethoxypyrimidin-4-yl)sulphanilamide, Infra red, electronic and X-ray diffraction study ©2011 ijCEPr. All rights reserved
INTRODUCTION Literature survey reveals that complexes of metal salts are more potent and less toxic in many cases as compared to
the parent drug[1]. These complexes are found to be interesting due to their biological application like antifungal[2],
antibacterial[3] activity. Large number of drugs has been used to synthesize the complexes with many metals with a
view to enhance their therapeutic action[4-6]. In continuation of our studies on the metal complexes of N1-(5,6
dimethoxypyrimidin-4-yl) sulphanilamide we report here Infra red ,electronic and X-ray diffraction study of Fe(II),
Co(II), Cu(II) metal chelates.
EXPERIMENTAL All the chemicals used in the present study were from BDH grade. Metal salts and solvents were used of reagent
grade. Metal ion solution and ligand solution in appropriate as desired and pH of resulting mixture were maintained
about 6.8 to 7.1 by putting alcoholic ammonia solution. The reaction mixture was refluxed for three hours by
keeping the round bottom flask on steam bath, The solid thus separated on cooling was filtered ,washed and dried in
vacuum desiccators over anhydrous CaCl2.The detailed preparation of complexes were discussed in earlier papers.
RESULTS AND DISCUSSION Infrared Spectral study
Important absorption frequencies of ligand and complexes along with their assignment are presented in the table-1.
The assignments are well supported by literature survey. The comparision of IR spectrum data of Fe (II), Co (II)
and Cu (II) complexes with ligand N1_
(5, 6-dimethoxypyrimidin-4-yl) sulphanilamide helps in determining bonding
pattern in the complexes.
Table-1: Infrared spectral data of the ligand (DMPS) and their metal complexes
IR spectra of ligand shows strong bond in the region 3238 cm-1
which is assigned to ν(NH2) Stretching vibration[7].
Ligand shows band at 3116 cm-1
assigned to ν(NH) stretching[8]. Band at 1010 cm-1
is due presence of (OCH3)
group in ligand[9].
Compound ν(NH2) ν(NH) ν(OCH3) ν(M-Cl) ν(M-N) ν(M-O)
DMPS 3238 3216 1010 - - -
[Fe(DMPS)2 Cl2] H2O 3296 3256 1010 340 459 553
[Co(DMPS) Cl2 2H2O] 3334 3248 1010 317 417 519
[Cu (DMPS) Cl H2O] Cl 3200-
3449
3200-
3449 1010 299 432 502
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Vol.2, No.1, 20-25 (2011)
Jitendra H. Deshmukh and M. N. Deshpande 21
The IR of Fe (II) complex shows strong band at 3406 cm-1
assigned to presence of lattice water in the
complex[10].In the IR Spectra of Fe (II) complex, ν(NH2) band shifted and observed at 3296 cm-1
indicating
formation of co-ordinate bond with metal. Similarly ν(NH) band in ligand is at 3216 cm-1
which shift to higher
frequency region 3256 cm-1
in complex. The additional bands around 459 cm-1
and 553 cm-1
are assigned to ν(M-N)
and ν(M-0) Stretching vibration, respectively. These bands were not observed in ligand. Co-ordination with the
chlorine atom is supported by the appearance of band in the far IR region at 340 cm-1
which may be assigned to
ν(M-Cl) linkage[11].
IR spectra of Co (II) complex shows intense bond at 3334cm-1
due to ν(NH2) band. This band in ligand observed at
3338 cm-1
. This band shifting indicates coordination of nitrogen with metal ion. In ligand ν(NH) observed at 3216
cm-1
but Co(II) complex it is shifted towards higher frequency and appears at 3248 cm-1.
In the far IR spectra region
of the complex, the bands at 417cm-1
and 519cm-1
observed can be assigned to ν(M-N) and ν(M-O) stretching
vibration respectively. Strong band at 3465 cm-1
indicates the presence of lattice water and ν(M-Cl) stretching
vibration is observed at 317cm-1
.
In the IR spectra Cu (II) Complex band at 3475 cm-1
is observed indicating the presence of coordinated water.
ν(NH2) and ν(NH) bands merge in the complex and appeared 3200-3449 cm-1.
Similarly low intense band is
observed at 432 cm-1
and 502 cm-1
due to formation of ν(M-N) and ν(M-O) bond. Band at 299 cm-1
indicates
formation of coordinate bond of chlorine with metal ν(M-Cl).
Electronic spectra
The electronic spectrum deals with transitions in UV and Visible region. The electronic spectra of the complexing
agent N1_
(5, 6-dimethoxypyrimidin-4-yl) sulphanilamide and their metal complexes were obtained from SAIF, IIT
Chennai. The ligand exhibit strong bands around 36000 cm-1
with a shoulder at 33,000cm-1
assigned to π→π* and
n→π* transitions respectively[12]. Different absorption bands and corresponding transitions are given in the table 2.
Table-2: Electronic spectral data in cm-1
and magnetic moment values of
Fe (II), Co (II) and Cu (II) complexes with DMPS
Electronic spectra of Fe (II) complex have different transitions and different absorbance peaks due to presence of the
metal ion in the complex. Fe (II) complex is d6 system having four unpaired electrons. Therefore complex is
paramagnetic. Due to presence of the d-d transitions some different peaks are observed in electronic spectra of the
complex than the electronic spectra of the ligand. In electronic spectra of Fe(II)complex low intensity bands 23774
cm-1
, 21505 cm-1
and 16606 cm-1
no specific assignments are made[13].Complex may acquire octahedral geometry
outer hybrid orbital are used to form co-ordinate bond between donor atom of ligand and the central metal ion. The
magnetic moment shown by metal complex is 5.05 BM. Electronic spectra of Co (II) complex exhibit three
absorbance peaks at 22779 cm-1
, 16339 cm-1
and 14925 cm-1
. These absorbance maxima due to 4T1g(F)→4
T1g(P), 4T1g(F)→4
A2g(F) and 4T1g(F) →
4T2g, transitions respectively characteristics of the octahedral geometry around
Co(II)metal ion[14]. Magnetic moment is found to be 3.85 BM. Electronic spectra of Cu (II) complex have three
bands at 22727 cm-1
, 17921cm-1
and 14513cm-1
indicating the transition between the ligand to copper metal ion.
The geometry of the complex is tetrahedral. Magnetic susceptibility indicates the presence of one unpaired electron.
The presence of above bands in electronic spectra of Cu (II) complex indicates 4A2→4T1g transition and also the
transitions due to charge transfer. Magnetic moment for the complexes is found to be 2.08 BM.
Electron Spin Resonance study
Compound Absorbance band
cm-1
Transitions Magnetic
Moment (BM)
[Fe (DMPS)2Cl2] H2O 23774, 21505,
16606
No specific assignment are
made
5.05
[Co(DMPS)Cl2 2H2O] 22779, 16339,
14925
4T1g(F)→4
T1g(P) 4T1g(F)→4
A2g(F), 4T1g(F) →
4T2g,
3.85
[Cu (DMPS) Cl H2O] Cl 22727, 17921,
14513
4A2→
4T1g
charge transfer
2.08
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Vol.2, No.1, 20-25 (2011)
Jitendra H. Deshmukh and M. N. Deshpande 22
The ESR spectrum of the Fe (II), Co (II) and Cu (II) complexes was recorded at room temperature using
tetracyanoethylene radical as ‘g’ marker. The H|| and H⊥ values were measured from the spectrum and used to
calculate the g|| and g⊥ values by using the formula are given in the table-3.
ESR spectra of Fe (II) complex in polycrystalline state shows two peaks. From the observed ESR of Fe (II)
complex, the ‘g’ value is 2.203 which is less than 2.3 indicating the coordinate bond between donors atoms of ligand
with metal ion have partial covalent character.From ESR spectra of Co (II) complex, g|| obtained is 1.84 and g⊥ is
2.36. gav value for Co (II) complex is found to be 2.18 which is less than 2.3 indicating covalent character of the
metal ligand bond[15].
ESR spectra of Cu (II) complex show two peaks. One of intense absorption peak at high field and the other of less
intensity peak at low field. From these two peaks, g|| and g⊥ have been calculated. The gav value of Cu (II) is 2.34
which are more than 2.3 indicating the presence of ionic character in the complex.
Table-3: ESR spectral data of Fe(II), Co(II) and Cu(II) complexes with DMPS
ESR spectral
parameters [Fe (DMPS)2Cl2] H2O
[Co(DMPS)Cl2
2H2O]
[Cu (DMPS) Cl H2O]
Cl
g|| 1.78 1.84 1.91
g⊥ 2.40 2.36 2.55
gav 2.203 2.18 2.34
X-ray diffraction Study
The crystal lattice parameters of Fe (II), Co (II), Cu (II) complexes with DMPS were found out by X-ray diffraction
powder method. The X-ray diffraction of complexes was recorded in the range 200 to 80
0 on 2θ value. The major
refluxes were measured and the corresponding d-values were obtained. An independent indexing for each of these
refluxes was carried out by least square method[16]. The miller indices (h k l) were calculated and refined using
Back-cal program by computational method and data has been summarized in the following tables.
Table-4:Cell data and crystal lattice parameters for [Fe (DMPS)2Cl2] H2O complex
a (A0) = 30.052210 ± 0.035028 Volume (A0)3 = 19578.25
b (A0) = 30.052210 ± 0.032651 Dcal = 1.0387 g/cm
3
c (A0) = 25.031710 ± 0.093737 Dobs = 1.2985 g/cm
3
Standard deviation=0.049395 Z = 16
= 4.9% Crystal system = Hexagonal
α=90° β=90° γ=120° Porosity (%) = 20
I/Io Dobs Dcal h k l
11 4.362115 4.378966 6 -1 2
49 3.902708 3.919956 7 -1 1
65 3.789483 3.783258 7 -1 2
38 3.682723 3.677650 8 -3 1
32 3.533657 3.532010 7 -3 4
72 3.486692 3.467847 8 -2 2
33 3.115265 3.110134 7 -1 5
16 2.847374 2.851161 10 -3 2
51 2.455439 2.451257 9 2 3
100 2.196959 2.198960 10 3 1
28 2.049400 2.051326 6 6 7
27 1.990465 1.990256 9 6 0
19 1.882965 1.885334 9 2 9
17 1.798814 1.798980 8 8 4
)g2g(3
1g ||av ⊥+=
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Vol.2, No.1, 20-25 (2011)
Jitendra H. Deshmukh and M. N. Deshpande 23
16 1.662780 1.662464 10 8 1
The cell data and crystal parameters of Fe (II) complex is given in the table indicates that the complex have
hexagonal crystal system.
Table-5: Cell data and crystal lattice parameters for [Co (DMPS)Cl2 2H2O] complex
a (A0) = 30.059050 ± 0.047643 Volume (A
0)
3 = 19552.39
b (A0) = 30.0659050 ± 0.045038 Dcal = 1.2940 g/cm
3
c (A0) = 24.970650 ± 0.053964 Dobs = 1.3988 g/cm
3
Standard deviation = 0.0050235 Z = 32
= 0.50% Crystal system = Hexagonal
α= 900 β= 90
0 γ =120
0 Porosity (%) = 7.49
I/Io Dobs Dcal h k l
20 3.902708 3.921863 7 -1 1
9 3.646795 3.641020 4 -2 6
7 3.631565 3.611176 8 -2 0
25 2.675749 2.677321 5 0 8
14 2.478043 2.479616 8 -1 7
23 2.411530 2.414165 8 4 2
14 2.308712 2.309016 10 0 5
13 2.214781 2.215209 10 1 5
9 2.019460 2.022433 10 4 3
16 1.935065 1.936715 9 6 3
39 1.870411 1.871525 10 2 8
16 1.702111 1.700870 9 8 4
19 1.572922 1.569306 10 9 2
Cell data and crystal lattice parameters of Co (II) complex attributed to hexagonal crystal system.
Table-6: Cell data and crystal lattice parameters for [Cu (DMPS) Cl H2O] Cl Complex
a (A0) = 21.878380 ± 0.015787 Volume (A
0)
3 = 14362.44
b (A0) = 23.412220 ± 0.041224 Dcal = 0.8558 g/cm
3
c (A0) = 28.039510 ± 0.054712 Dobs = 1.0963 g/cm
3
Standard deviation = 0.0030459 Z = 16
= 0.3% Crystal system = Orthorhombic
α= 900 β= 90
0 γ = 90
0 Porosity (%) = 24.05
I/Io Dobs Dcal h k l
8 4.087049 4.098696 5 2 0
27 3.789483 3.793231 3 5 2
17 3.533657 3.528988 6 0 2
12 3.440960 3.434774 3 5 4
11 3.396491 3.397027 6 0 3
100 2.878237 2.873455 7 2 3
7 2.675844 2.680874 7 3 4
10 2.524552 2.526895 6 0 8
11 2.478043 2.477680 8 4 0
10 2.369269 2.368666 6 0 9
11 2.232888 2.232415 8 6 1
7 2.128633 2.125735 10 2 2
10 1.935064 1.935126 9 3 8
Cell data and crystal lattice parameters of Cu (II) complex indicates that complex have orthorhombic crystal system.
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Vol.2, No.1, 20-25 (2011)
Jitendra H. Deshmukh and M. N. Deshpande 24
Cl
Cl
N
N
N
O
O
C H 3
C H 3
S O
O
N H
H
N
N
N
O
O
CH 3
CH 3
SO
O
N
H
H
H
Fe
H
OH 2
Mol. formula C24H30N8O9S2Cl2Fe; Mol.wt. = 765.64
Dichloro bis N1(5, 6-dimethoxypyrimidin-4-yl) sulphanilamide Fe (II) complex.
O
H
H
O
H
H
Cl
Cl
N
N
N
O
O
C H 3
C H 3
S O
O
N H
H
Co
H
Mol. Formula: C12H18N4O6SCl2Co; Mol. wt. = 476.32
Dichloro N1–(5, 6-dimethoxypyrimidin-4-yl) sulphanilamide diaquo Co (II) Complex
Cl N
S
O
O
N
N
N
H
O
O
CH3
CH3
H
H
O
HH
Cu
Cl
Molecular Formula: C12H16N4O5S Cl2Cu ; Mol. wt. = 462.82
Monochloro N1-(5,6-dimethoxypyrimidin-4-yl) sulphanilamide aquo Cu(II) Chloride Complex.
CONCLUSION All the complexes are paramagnetic. Electronic spectrum of each metal complexes produce intense peak at higher
wave number. From the foregoing observations, the suggested chemical structures for the prepared complexes
under investigation are as follows.
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Vol.2, No.1, 20-25 (2011)
Jitendra H. Deshmukh and M. N. Deshpande 25
ACKNOWLEDGEMENTS The author would like to thanks the Principal and Head Department of Chemistry, Science College, Nanded for
providing all necessary facilities. In addition, the author wishes to acknowledge Principal Yeshwant College,
Nanded for encouragement.
REFERENCES 1. Singh A. and Singh P., Indian J. Chem.,39A (2000)874
2. Sharma R.C. and Parashar R.K., J. Inorg. BioChem. ,32(1998) 163.
3. Adbel-Waheb Z.H., Mashaly M.M., Salman A.A., El-Shetary B.A. and Faheim A.A. Spectrachim Acta., 60
(2004)2861.
4. Reedijk J., Pure Appl.Chem. ,59(1987) 181.
5. Lochrer P.J. and Einhorn L.H., Ann.inten, Med .,100(1984) 704.
6. Bell R.A., Lock C.J.L., Scholten C. and Villiant J.F., Inorg. Chem. Acta, 274 (1998) 137.
7. Farinan J.A., Patel K.S. and Nelson L.O.; J. Inorg. Nucl. Chem., 38(1976) 77.
8. Mitu L. and Kriza Angela, Asian J. of Chem. , 19(1)(2007) 658.
9. Bellamy L.J., The infrared organic molecules,1958, John Wiley Sons, Inc. New York.
10. Nakamoto K., 1963, IR spectra Wiley New York .
11. Goldstein M. and Unworth D., Inorg. Chem. Acta., 4(1970)37.
12. Seetharama Rao T., Laxma Reddy K. and Lingaih Ind. Acad Sci. (Chem.Sci.) , 100(5)(1988) 333.
13. Anuradha G.H. and Chandrapal A.V., Ori. J. Chem., 23 (1) (2007) 287.
14. Mathew and Watson (1971).
15. Kivelson D. and Meinan R., J. Chem.Phys., 35 (1961) 149.
16. Stout G.H. and . Jensen L.H., 1968, X-ray structure determination a practical guide, MacMillan; New
York.
[IJCEPR-161/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 26-29
January-April, 2011
Preeti Nandkumar
Comparative Study between soda Lignin and soda Anthraquinone lignin in
terms of Physiochemical Properties of Ipomoea carena
Preeti Nandkumar Department of Applied Chemistry,
M. P. Christian College of Engineering & Technology, Bhilai – 490026.
E-mail: [email protected] Article History:
Received: 17 March 2011
Accepted: 27 April 2011
ABSTRACT
Non wood plants are more common as raw material where wood is scarce. In view of its easy availability of ipomoea carnea was
utilized as raw material.In this study soda lignin and soda anthraquinone lignin were studied.A comparision has been made
between the physico chemical properties and structural features of isolated lignin.These characterisation was done by Fourier
Transform Infrared specrometry(FTIR),Ultraviolet(UV)and High Perormance Liquid chromatography(HPLC).Nitrobenzene
oxidation was performed with the two types of lignin especially soda lignin and soda anthrquinone lignin.According to the FTIR
report, there is no significant difference in terms of functional groups that exists in both the lignin. HPLC results however
identified that in both the lignin samples the presence of vanillin and syringaldehyde was found.
Key words: Ipomoea carnea, lignin, anthraquinone, black liquor. ©2011 ijCEPr. All rights reserved
INTRODUCTION Ipomoea carnea is a common weed and locally known as BESHRAM. It is also known as bush morning glory which
is fast growing and attains optimum size in about a year’s time. Due to its high adaptability and resistance towards
adverse climatic conditions, it may grow in all types of climate and soils, marshy as well as dry. A large diffused or
straggling shrub with milky juice native of South America, the plant was originally used for making fence for the
road side fields, but due to its massive growth and rapid propagation it has grown rapidly in barren waste lands.
Plantations of ipomoea carnea may be undertaken in the month of June, July with the onset of monsoon[1].
Ipomoea carnea plant is poisonous to animals. Its leaves contain a polysaccharide-ipomus, one glucoside-
anthracene a gum-gelapin and saponin .Out of the two materials, one is soluble in water and the other is in ether.
Both polysaccharide and anthracene present in ipomoea carnea are water soluble poisons when enters into the
central nervous system it damages the respiratory track[2], the scarcity and the restricted supply of high quality pulp
and the rising price of utilities will force paper mills to adopt new technologies to conserve energy, minimum inputs,
keeping environmental aspects in view, much efforts have been directed towards finding a chemical pulping process
giving higher pulp yield coupled with economic and environmental considerations[3].
The process of producing cellulosic pulp from ipomoea carnea jacq requires delignification with sodium hydroxide
under pressure.This process frees the cellulosic fiber from ipomoea carnea and produces a large quantity of black
liquor that is discharged into surface water without effective treatment[4].Based on the study of ipomoea carnea as
lignocellulosic raw material for the pulp and paper industry, sodium hydroxide lignin extracted from soda pulping
has been compared with soda anthraquinone (AQ) lignin extracted from soda AQ pulping in this study. Lignin
extracted during the pulping process has so far not being investigated for its usefulness. Before its application can be
considered, knowledge of its structural characterization is required; this study represents such an effort.
Lignin is an amorphous polyphenolic material arising from an enzyme mediated dehydrogenates polymerization of
three major phenyl propanoid monomer which is coniferyl, sinapyl and p-coumaryl alcohol. The lignin structural
elements are linked by carbon-carbon and ether bond to form tridimensional network associated with the
hemicelluloses polysaccharide inside the cell wall. Lignin is usually insoluble in all solvents and can only be
degraded by physical or chemical treatment. During the chemical pulping process at high temperature and high
pressures degradation of lignin occurs and dissolves into the spent liquor. The delignification reactions involved the
cleavage of non phenolic β-O-4 linkage, phenolic α-O-4 linkage and releasing from the associated by the
polysaccharide. Addition of small quantity of anthraquinone to the alkaline pulping process increases lignin
removal by promoting cleavage of interunit bonds in the lignin molecules that are not cleaved in the absence of
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Vol.2, No.1, 26-29 (2011)
Preeti Nandkumar
27
anthraquinone. Anthraquinone helps to minimize recondensations reactions by reacting with the carbohydrates to
increase lignin removal during pulping process[5,6].
This study was conducted to characterize soda lignin and soda anthraquinone lignin in terms of their
physicochemical properties and their structural features. The objective is to determine that adding anthraquinone to
the pulping process changes the properties of the lignin produced. Complimentary destructive nitrobenzene
oxidation and non destructive Infrared (IR), Ultraviolet (UV) and high performance liquid chromatography (HPLC)
to evaluate the cross linked lignins and their linkages to cell wall polysaccharide.
MATERIAL AND METHODS Raw material used for the laboratory experiment is ipomoea carnea jacq. The sample were collected, cleaned,
chipped and screened. The screened chips were used for the experiment. About 500 gms of screened chips of
ipomoea carnea jacq was pulped by soda pulping and soda anthraquinone pulping in a 20 liter stainless steel rotatory
digester unit with 25% NaOH (cooking liquor) in 3hrs at a maximum cooking temperature of 170oC at a pressure of
10psi with a cooking liquor to ipomoea carnea ratio of 10:1 by weight. For soda AQ pulping 0.1% anthraquinone
was added to the soda pulping system.
The soda and soda anthraquinone lignin were precipitated from the black liquor by acidifying it to pH 50mg of dry
soda lignin or soda anthraquinone lignin was added to 7ml of 2M, NaOH and 0.4ml of nitrobenzene in a 15 ml steel
autoclave. The autoclave was sealed tightly with a screw cap fitted with Teflon gasket and heated to 165oC for 3hrs
in an oil bath. After heating the autoclave was cooled quickly by immersion in ice water. The soda lignin mixture
was transferred to a liquid-liquid extractor for continuous extraction with 10ml chloroform to remove any remaining
nitrobenzene reduction products and excess nitrobenzene. The oxidized mixture was acidified with conc.HCl to pH
3-4 and then extracted with 20ml chloroform. The chloroform was removed by using a rotatory evaporator at 40oC
under reduced pressure to obtain nitrobenzene oxidation mixture which was used with a stock solution for further
analysis.
Table-1:Yield and molar ratio of degradation products of soda lignin and soda AQ lignin by nitrobenzene oxidation
Oxidation
peak
Oxidation product Soda retention
time
Liquid
yield %
Soda AQ
retention time
Lignin yield
%
A p-hydroxy benzoic acid(H1) 4.29 4.98 4.3 0.64
B Vanillic Acid (V1) 5.23 3.98 5.28 5.65
C Syringic Acid (S1) 5.55 4.74 5.6 4.92
D p-hydroxy benzaldehyde(H2) 6.53 26.54 6.58 15.97
E Vanillin (V2) 8.44 30.33 8.51 36.86
F p-coumaric acid(B) 10.13 26.54 10.17 31.95
G Syringaldehyde (S2) 12.38 2.84 12.42 3.69
Molar
Ratio
S/S
V/S
H/S
1
5
4
1
5
2
S=S1+S2, V=V1+V2, H=H1+H2
Table-2: IR Stretching frequencies
S.No. Type of Bond Stretching frequencies Intensity
1. O-H Bond 3430-3400cm-1
Strong,
broad
2. C-H Bond( in methyl group) 2940-2930cm-1
Medium
3. C-O ( in carbonyl compounds) 1720-1660cm-1
Strong
4. C-O (in conjugated carbonyl compounds with aromatic ring 1712-1702cm-1
Medium
5. Aromatic ring 1609-1604cm-1
1516-1510cm-1
1426-1422cm-1
Strong
6. C-H Bond (bending vibrations from aromatic group ) 1470-1460cm-1
Medium
7. C-O ( in syringyl group ) 1330-1325cm-1
1117-1115cm-1
Weak
Medium
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Vol.2, No.1, 26-29 (2011)
Preeti Nandkumar
28
8. C-O(in syringyl and guaiacyl group) 1220-1215cm-1
Strong
9. C-O(in guaiacyl group) 1158-1155cm-1
1038-1030cm-1
Strong
10. Bending vibrations inside aromatic plane for guaiacyl ring 1038-1030cm-1
Strong
11. C-H deformation and ring vibration 838-834 cm-1
Medium
High performance liquid chromatography (HPLC) was used to analyze the nitrobenzene mixture. Stock
solution(0.25ml) was pipette into 25ml volumetric flask and made up to volume with acetonitrile:water (1:2
v/v).forty micro liter of the filtrate was injected into an HPLC system equipped with hypersil bond C18 column to
identify oxidation product. A 1:8 mixture of acetonitrile: water containing 1% acetic acid was used as an eluent with
a flow rate of 2ml min-1.the eluent was monitored with an UV (ultraviolet) detector at 280nm.IR spectra were
recorded with a Perkin Elmer spectrophotometer for each sample. KBr pellets were prepared containing 1% finely
ground sample. For UV spectra-A Hitachi spectrophotometer was used to obtain the results. Prior to the
analysis,5mg samples were dissolved in 10ml 90% (v/v) dioxane:water.The sample was then measured its
absorbance for range of 210 to 350 nm.
Fig.-1
RESULTS AND DISCUSSION Yield of soda anthraquinone lignin was much higher as compared with the yield of soda lignin. It was found that
anthraquinone lignin was 9.6% high than soda lignin. The amount of solubilized lignin in soda AQ black liquor is
higher because anthraquinone serves as a catalyst for the soda pulping process.AQ has a marked catalytic effect on
the delignification.AQ acts in a redox sequence and cycles between its oxidized and reduced forms. The oxidized
AQ form reacts with quinine methide segments of the lignin polymer to increase the rate of delignification.
Nitrobenzene oxidation is one of the standard methods for analyzing lignin by chemical degradation technique in
order to gain information about the composition of the original polymer. The production of aromatic aldehyde upon
oxidation of lignin with alkaline nitrobenzene takes place. Three monomeric lignin units’ i.e. p-hydroxyphenyl (H),
guaicyl (V) and syringyl (S) based on the amount of their degradation product. The degradation product of
syringaldehyde and vanillin were analysed.Syringaldehyde was found to be predominant followed by vanillin as a
second major degradation product. HPLC chromatogram for soda lignin and soda AQ lignin are similar. In general
the S: V: H ratio for both syringaldehyde and vanillin. The lignin’s are about the same which is 1:5:4 for soda lignin
and 1:5:2 for soda AQ lignin.
IR spectra (Fig.-1) of soda lignin and soda AQ lignin precipitates have a strong and broad band at 3406cm-1
which is
a characteristic of OH group or phenolic compound from soda AQ lignin. The band width and strength could be due
to moisture in the sample, since the OH vibration of water usually is very broad. The clear peak at 2934-2844 cm-1
for the soda AQ lignin is attributed to the vibration of a Methoxyl (-OCH3) group while slightly different values
were observed for soda lignin (2936-2844 cm-1
).the band at 1462 cm-1
is assigned to CH stretching of methyl or
methylene groups and the broad medium band at 1712 cm-1
is due to conjugated carbonyl stretching. The three
bands at 1606, 1515 and 1425 cm-1
are characteristics of aromatic rings due to aromatic vibrations and the band at
832cm-1
indicates CH deformation and ring vibrations. The bands at 1329 cm-1
for soda AQ lignin and 1328 cm-1
for
soda lignin may be due to the vibration of C aryl-O in syringyl derivatives. The bands at 1328-1329cm-1
and 1216
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Vol.2, No.1, 26-29 (2011)
Preeti Nandkumar
29
cm-1
corresponds to a syringyl units and the small bands at 1033-041cm-1
are assigned to guaiacyl units of lignin
molecules
CONCLUSION Addition of anthraquinone to the pulping process does not affect the quality of lignin precipitated from soda black
liquor; even though it nearly doubles the amount of lignin precipitated from black liquor. Rate of delignification was
higher with 0.1% addition of anthraquinone.
The production of aromatic aldehyde upon oxidation of lignin with alkaline nitrobenzene takes place the product
yielded are vanillin and syringaldehyde. Molar ratio of syringyl and guaicyl unit varies from species to species even
in the same genus and exerts its influence on rate of delignification. Higher the ratio better is the material from
delignification and from processing lignin containing spent liquor points of view.
REFERENCES 1. Ipomoea Carnea; The Wealth of India CSIR publication, raw material, 5 (1950) 58.
2. Nair Preeti , Shukla R.N., Indian Journal of Applied and Pure Bio., 19(2) (2004)189.
3. Venica, A., C.L.Chen and J.S.Gratzl, Delignification of hardwoods during alkaline pulping; reactions,
mechanisms and characteristics of dissolved lignin’s during soda aqueous pulping of poplar, Tappi
proceeding, pp.503, 1989.
4. Sucking I.D., The role of anthraquinone in sulphite pulping TAPPI wood and pulping chemistry Tappi
proceedings, pp. 503(1989).
5. Lin, S.Y., and C.W.Dence, Method in Lignin Chemistry, Springer pp 65-67, 71-73, 75-80.,(1992).
6. Sun, R.C., J. Tompkinson and G.L.Jones ,Fractional Characterization of Ash by Successive Extraction with
Organic Solvents.,68 (2000)111.
[IJCEPR-154/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 30-33
January-April, 2011
Manju Chaudhary
Use of Millet Husk as a Biosorbent for the Removal of chromium and
Manganese Ions from the Aqueous Solutions.
Manju Chaudhary Department of chemistry, Siddhi Vinayak College of Science and Higher Education, Alwar, Rajasthan.
E-mail: [email protected] Article History:
Received:28 January 2011
Accepted:5 February 2011
ABSTRACT
Movement of heavy metal ions into the water sources has made it unfit for consumption. Rural areas situated near the industrial
area are facing the problem of contaminated water. Present work is an effort to develop a self sufficient, easy to assemble and eco
friendly system for the removal of heavy metal ions from the water. Millet husk readily available in the rural areas of eastern
Rajasthan has been tried and tested for the removal of chromium and manganese ions from the aqueous system. The results are
exiting and indicate that it could be used for the removal of heavy metal ions from the water. A column of 12 inches height and 2
inches diameter of pretreated millet husk was used as a biosorbent. This column is sufficient to treat a 50 ml solution (0.012g/L)
of chromium ions and 65ml solution (0.001g/L) of manganese ions. The rate of flow of the solution was kept 2ml/min at room
temperature and Ph3-4.
Key words: biosorbent , millet husk, biosorption, Chromium ions, manganese ions. ©2011 ijCEPr. All rights reserved
INTRODUCTION Growing industries gave good job opportunities and pace to the economic growth of India and at the same time pose
serious threat to the environment. Water being most important part of industries and environment has been affected
most. Quality of water in the nearby areas of the industrial growth is facing a serious problem of heavy metal
contamination. Surface water as well as underground water is equally affected. About 20 metals have been identified
as toxic to human health and out of this half are emitted into the environment in quantities that pose risk to human
health.
A number of chemical and physical processes are available for the removal of these toxic ions from the water
samples. Of all the available processes which can remove these toxic ions from water have some or the other
technical or economical problem. A study of such systems and their difficulties has been reported [1]. Therefore
some methods which are environment friendly and low cost are desirable. A study on the different types of
biosorbents and their probable use has been presented by Volesky et al [2]. Here is a useful method which in recent
years has gained a lot of attention.
Biosorption can be defined as a ‘non directed physico-chemical interaction that may occur between
metal/radionuclide species and microbial cells [3].A variety of micro organisms as biosorbents have been identified
and reported[4-7] for the removal of heavy metal ions. Some plant wastes like obtained from agriculture farms have
also been reported to be efficient biosorbents[8-15].Removal of chromium[16-18], iron[16,19], lead[19,20] and
copper [19-21])have also been studied and reported .
In the present paper the biosorption of lead, copper and cadmium on the two biosorbents that are available in plenty
in southern Rajasthan are reported.
MATERIALS AND METHOD Preparation of biosorbent The biowaste that is the husk of millet and oat have been procured from the nearby village. The husk was washed
thoroughly with water to remove sand etc. Then it was treated with dilute HCl to remove natural colour of the husk
and then again washed with demineralised water. Thoroughly washed husk have dried in an hot air oven at about 60 0C for 24 hours. The properly dried husk is now ground to a fine powder and kept in air tight jars and taken as and
when required.
A steel column of 20 inches height and 2inches diameter has been taken. The biosorbent was filled into the column
till a height of 12 inches only. All the experiments have been carried out using this column only.
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Vol.1, No.2, 30-33 (2011)
Manju Chaudhary 31
Preparation of the solutions: Standard solutions (1000mg/L) of chromium (VI) and manganese (VII) were prepared
in the laboratory by dissolving potassiumdichromate and potassium permanganate in doubly distilled water. Stock
solutions were kept in covered and labeled bottles. These stock solutions were used after further dilution according
to the requirement. The pH of the solutions was adjusted using NaOH and HCl.
Method
The test solution containing metal ions was allowed to flow through the prepared column of 12 inches height and 2
inches diameter. The flow of test solution was maintained by using a burette. The flow rate was maintained 3 ml/min
for all the experiments except when effect of variation of the rate of flow was studied.
A clean tumbler properly washed with deionised water was used to collect the percolated water. All the test
solutions were examined before and after the treatment using atomic absorption spectrophotometer.
Sorption efficiency (%) of the biosorbent was calculated in terms of amount of metal ion adsorbed per gram of the
biomass using following formula.
Q = (C0-C)/M X V
Sorption efficiency (%) = C0 – C /C0 X 100
Where Q is the amount of metal ion biosorbed per gram of the biomasss mg/g.C0 and C are the initial and final
concentrations of the test solution. V is the volume of the test solution; M is the mass of the biomass (g).
RESULTS AND DISCUSSIONS Effect of pH on the sorption capacity of the biosorbent
Biosorption of metal ions on to the surface of the biosorbent is greatly influenced by the pH of the solution. It is
expected the sorption of the metal ions increase with increase in the pH of the solution. The possible reason may be
that at lower pH, H+
and H3O+ ions compete with the metal ions for adsorption onto the surface of biosorbent. At
higher pH less no of H+ ions are there to occupy the adsorbent sites are available for the metal cations. The pH of the
solution was varied from 2 to7. The pH of the normal water is around 7 therefore the pH 7 is chosen to conclude the
adsorption capacities of the biosorbent. The case is different with the chromium ions.Chromium ions are available as
anions CrO32-
or HCrO3- in the solution therefore they showed a different character. It has been found that
biosorption of chromium decrease with increase in the pH of the solution. Probable reason may the presence more
number of positive sites on the biosorbent at lower pH which can adsorbe chromate ions. Similar is the case with the
manganese ions.
It is clear from the following table that the extent of adsorption decreases with increase in the pH of the solution for
manganese ions and chromate ions. The results are shown in table1.
Table-1
Effect of Rate of flow of solution through column
Rate of flow of solution through the biosorbent column effect the net percentage removal of metal ions from the
solution. If the solution travels quickly down the column then the metal ions get less time to adhere to the surface of
the biosorbent. Similarly if the rate of flow through the column is slow than the metal ions will get sufficient time to
get adhere to the surface of the biosorbent. Therefore the effect of rate of flow on the percentage removal of the ions
from the test solutions must be considered and studied. Following figures illustrates that with the increase in the
contact time between the adsorbent and the metal ions from test solutions percentage removal of the ions is also
increased. The results are shown as below in table 2.
pH Percentage removal of Cr(VII) Percentage removal of Mn(VII)
1
2
3
4
5
6
7
78%
88%
65%
49%
18%
8%
2%
75%
90%
82%
32%
12%
2%
0
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Vol.1, No.2, 30-33 (2011)
Manju Chaudhary 32
Table-2
Rate of flow of solution
(mL/ min)
Percentage removal of
Cr(VII)
Percentage removal of
Mn(VII)
2
4
6
8
10
88%
72%
66%
62%
53%
90%
53%
48%
34%
20%
Effect of height of column
Keeping the diameter of the column fix and varying the height of the column it is seen that the sorption of ions is
greatly influenced. Increase in the height of the column means increase in the number of adsorption sites. Therefore
the extent of sorption is to increase with the increase in the height of the column. Height of the column is varied
from 8inches to 14 inches. Results are shown below in Table 3.
Table-3
Effect of initial concentration of the metal ion solution
Initial concentration of metal ions in the solutions affects the rate sorption of ions on the sorbent surface. Initially the
rate of adsorption increase with increase in initial concentration of the ions in solution. This is due to large number
of adsorption sites available for few metal ions in the solution. On further increase in the concentration the rate of
adsorption nearly become stable when number of metal ions and number of adsorption sites become equal. Beyond
this concentration there is no increase in the rate of adsorption. The initial concentration of metal ions taken for the
study is 20 mg/L each. The equilibrium is reached at about 80 mg/L and then after no further increase in the rate of
adsorption is noticed. The results are shown below in table 4.
Table-4
Initial concentration of the metal
ions(mg/L)
Metal uptake of Cr(VII) (mg/g) Metal uptake of Mn(VII) mg/g
20
40
60
80
100
2.5
4.8
5.4
6.8
7.2
1.5
3.6
5.2
6.8
6.8
CONCLUSION Biosorption capacities of Pennisetum typhoideum (millet husk) are found enough for the removal of chromium and
mangenese ions from the waste water. The optimum ph range for the purpose lies in the acidic range. There is a
need to study the system in more detail and find the practical utility. Widely available and ecofriendly biowastes
like husk of millet and oat can serve the purpose of water purifier in the rural areas.
ACKNOWLEDGEMENTS I am thankful to Ms Seema Bhardwaj and Ms Akansha Arora for helping me in editing and formatting of the paper.
My sincere thanks are due to the college and chemistry department who provided me the laboratory facility.
Height of column in inches
(diameter 2 inches in all the
cases)
Percentage removal of Cr(VI) Percentage removal of Mn(VII)
2
4
6
8
10
12
23%
35%
50%
64%
76%
88%
18%
34%
52%
68%
83%
90%
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Vol.1, No.2, 30-33 (2011)
Manju Chaudhary 33
REFERENCES 1. Alhalya N., Ramchandra T.V., Kannamadi R. D., Res.J.Chem.Environ., 7 (2003) 71.
2. Volesky B. and Holan Z. R., Biotechnol Prog., II (1995) 235.
3. Shumate S. E., Strankberg G. W., Comprehensive Biotechnology; Pergamon Press, New York (1985) 235.
4. Kapoor A., Virarangahavan T., Bioresour.Technol, 63(1998) 109.
5. Kar R. N., Sahoo B. N., Shukla C. B., Pollut.Res.,11 (1992) 1.
6. Kolishka T., Galin P., Z.Naluforsch, 57 (200) 629.
7. Loderro P., Cordero B., Grille Z., Herror R.,Sastre de Vicente ME, Biotechnol.Bioeng., 88(2004) 237.
8. Sarin V. and Pant K.K., Bioresour.Technol., 97 (2006) 15.
9. Ricordel S., Taha S., Cisse I. and Dorange G., Sep.Purf.Technol., 24 (2001) 389.
10. Selwakumari S., Murugan M., Pattabi S., Sathish Kumar M., Bull.Environ. Cont. Toxicol., 69(2001) 195.
11. Gangsun and Weixing Shi, Ind. Eng.Chem.Res., 37(4) (1998) 1324.
12. Espinola A., Adamiuan R. and Gomes L.M.D., Waste Treat Clean Technol.Proc., 3(1999) 2057.
13. Iqbal M., Saeed A. and Akhtar N., Biores. Technol., 81(2002) 153.
14. Tee T. W. and Khan R.A.M., Environ.Technol. Lett., 9(1988) 1123.
15. Orhan Y. and Buyukgungar H., Water Sci.Technol., 28(1993) 247.
16. Ahalya N., Kanamadi R.D. and Ramachandra T. V., J.Environmental Biology 28(4) (2007) 765.
17. Vinodini V., Anabarasu and Nilanjana Das, Int. J.of natural products and resources, 1(2)(2009) 174.
18. Saifuddin M., Nomanbhay and Kumaran Paanisamy, Environmental Biotechnology, 8(1) (2005).
19. Fan Z., Xiaotao J., Int. J. Chem., 4(2002) 34.
20. Haluk C., Ulki Y., Water S. A., 27 (2001) 15.
21. Muraleedharan T.R., Venkobachr C., Biotechnol.Bioeng, 35(1990) 320.
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 34-39
January-April, 2011
P.J.Puri et al.
Study Regarding Lake Water Pollution with Heavy Metals in Nagpur City
(India)
P.J. Puri*,1
, M.K.N. Yenkie1, S. P. Sangal
2, N.V. Gandhare
2 and G. B. Sarote
3
*,1Department of Chemistry, LIT, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur - 440 001, 1 Department of Chemistry, LIT, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur - 440 001, 2Department of Chemistry, Nabira Mahavidyalaya, RTM, Nagpur University, Katol - 66302 3Regional Forensic Science Laboratory, Dhantoli, Nagpur – 440 012
E-mail: [email protected] Article History:
Received:6 January 2011
Accepted:20 January 2011
ABSTRACT This paper is intended to be a study concerning water pollution with heavy metals in Nagpur City, Maharashtra, India. The levels
of the occurrence of heavy metals like cadmium (cd), iron (Fe), zinc (Zn), arsenic (As), mercury (Hg), lead (Pb) and chromium
(Cr) were estimated in Futala, Ambazari, Gandhisagar and Gorewada lake, within Nagpur city, for the session January to
December 2008. Sampling points were selected on the basis of their importance. The monitoring was made over a period of one
year comprising of three seasons; summer, winter and rainy season respectively. The study demon started gradual increase in
pollution input of heavy metals in studied lakes. The yearly variation in the concentration of heavy metals had definite upward
trends. Present study revealed that dissolved constituents of Fe, Pb, Zn and Cr were above ranges of unpolluted water indicating
their contamination throughout the season in cases of Pb, Fe and Zn and occasional for As and Hg. The metals Zn, Fe, Cd, Ni
almost remained in natural level while arsenic (As) was always below the detection limit of 0.0001ppm. The Futala, Ambazari
and Gandhisagar except Gorewada lake could be identified as probable area of contamination of these metals. The average levels
of metals in studied lakes followed the order Zn > Cr > Fe > Cd > Pb > Hg > As.
Keywords: Heavy metals, pollution, Lakes, Water Quality. ©2011 ijCEPr. All rights reserved
INTRODUCTION Heavy metals are important environmental pollutants and their toxicity is a problem of increasing significance for
ecological, evolutionary, and environmental reasons[1]. Heavy metals have played great roles in genesis of present
day civilization. In ancient times, the wealth of Emperors and Kings was attributed to the possession of metals like
iron, gold, silver copper etc. in different forms. Still today, the dependence of heavy metals has not decreased as
these are very commonly used in agriculture, medicine, engineering etc. The magnitude of danger of environmental
pollution by heavy metals was probably for first time realized with the Minimata disaster in Japan, where thousand
of peoples suffered with mercury poisoning after consuming the fish caught in Minimata Bay. The bay got
contaminated with mercury released from vinyl chloride plant between 1953 and 1960[2]. Similarly, it was also
reported in Japan in 1955 that cadmium caused itai-itai Byo’ disease in human beings, mainly in women over forty.
This was due to high level of cadmium in local foodstuffs attributable to irrigation water from soil heaps of an
abandoned mine. Minimata raised its ugly head once again, not in Japan this time, but in fishing communities of
Amazon rain forest. Heavy metal pollution has been worked out in recent days[3-13]. Thus in view of this widely
used practice; it was of interest to undertake further investigation on these lines. The purpose of study is to
promoting and coordinating activities in the field of environmental chemistry as well as health related water
microbiology and hygiene.
MATERIALS AND METHOD Physicochemical characteristics (only pH, conductivity and turbidity), of water samples were determined within
twenty four hours of the collection of water samples using standard methods[14]. Total trace metals were
determined in acidified water samples after pre concentration by atomic absorption spectrophotometric methods.
The metals Fe, Pb, Hg, Cu, Zn, and As were estimated using Atomic Absorption Spectrometer (Make - GBC
Australia, Model GBC 932). Air-acetylene flame technique was used for all these metals and hydride generator
technique was used for arsenic. For dissolved metals, water samples were preserved by adding HNO3 and filtered
through Millipore filtering unit. One liter of the filtered sample was evaporated to dryness and digested with HNO3,
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Vol.2, No.1, 34-39 (2011)
P.J.Puri et al.
35
HCIO4 mixture. The digested samples were made up to the volume and dissolved metals were analyzed by AAS.
Iron, manganese and zinc were subjected to ten-fold concentration by evaporation. Cadmium, chromium, copper and
lead were concentrated by complexing with ammonium pyrrolidine dithiocarbamate (APDC) and subsequently
extracting the complex in methyl isobutyl ketone (MIBK). Iron, manganese and zinc in aquatic solution and
cadmium, chromium, copper and lead in organic solution, were determined by atomizing respective solutions in
Atomic Absorption Spectrometer. The instrument was operated in flame mode using air as oxidant and acetylene as
fuel Operation parameters were optimized for maximum response. Background correction was made for lead and
cadmium and flame rich in acetylene was used for chromium determination. Water samples were analyzed by both
classical and automated instrumental methods as appropriated in standard methods for analysis of water and waste
water[14]. All reagents used were of analytical grade and instruments pre-calibrated appropriately prior to
measurement. Replicate analyses were carried out for each determination to ascertain reproducibility and quality
assurance
RESULTS AND DISCUSSION Seasonal variations were noted in the physicochemical properties of studied lake water Different properties like pH,
EC, Temperature, HCO3- and
conductivity showed maximum values during summer, while minimum values were
recorded during autumn season. The observed trend could be attributed to the evaporation of water from studied lake
(Gandhisagar, Ambazari, Futala and Gorewada Lake) during summer and subsequent due to precipitation and runoff
from catchment area during rainy season[15-16]. High pH values in all studied lakes during summer could be
ascribed to increased photosynthetic assimilation of dissolved inorganic carbon by planktons[17]. A similar effect
could also be produced by water evaporation through loss of half bound CO2 and precipitation of mono-
carbonate[18]. The alkaline pH and high alkalinity of Futala, Ambazari and Gandhisagar lake water might be due to
use of detergents by neighboring population for washing of cloths, vehicles, and utensils. Higher alkalinity in Futala,
Ambazari and Gandhisagar indicated the potential susceptibility of these water bodies for eutrophication. Lake water
bodies with alkalinity values above 100 mgL-1
is considered nutritionally rich [19] and on the basis of this
observation most of lakes in Nagpur city could be considered prone to eutrophication problems.
Seasonal variation in different heavy metal concentration in water from Futala, Ambazari, Gandhisagar and
Gorewada lake are presented in Table 1-4. Graphical representation of seasonal variation in different heavy metal
concentration in water from Futala, Ambazari, Gandhisagar and Gorewada lake are presented in Figure1-4. Figure 5
represents map showing Gandhisagar, Ambazari, Gorewada and Futala lake, Nagpur (MS) India. The concentration
of heavy meals in water of studied lakes remained below toxic limits with few exceptions. A remarkable high
concentration of (Fe) iron, ranged from 0.022 mgL-1
to 0.035 mgL-1
, 0.014 mgL-1
to 0.031 mgL-1
, 0.025 mgL-1
to 0.031 mgL-1
and 0.012 mgL-1
to 0.016 mgL-1
in Futala, Gandhisagar, Ambazari and Gorewada lake
respectively. The Fe content indicated that this metal was abundant in soil and rocks of catchment area from where
the water reaches to these lakes. As regards the effect of season on heavy metals concentration in water of Futala,
Ambazari and Gorewada lakes, concentration of metals like Cd, Cr, Fe, Ni, Pb, Zn, Hg were maximum during
summer and rainy season while minimum concentrations were observed during autumn season. This trend could be
attributed to the evaporation of water from lakes during summer and subsequent dilution due to precipitation and run
off from catchment area during rainy season[20]. The variation of level of occurrence of heavy metal in Ambazari,
Futala and Gandhisagar lake were found different from each other due to the variation of the solubility of the
existing forms of metal in water as well as their availability in the immediate environment. Among metals the level
of Zinc ranged from 0.025 mgL-1
to 0.0.048 mgL-1
, 0.016 mgL-1
to 0.041 mgL-1
, 0.021 mgL-1
to 0.032 mgL-1
and 0.012 mgL-1
to 0.021 mgL-1
in Futala, Gandhisagar, Ambazari and Gorewada lake respectively. Arsenic (As)
level was always less than 0.1ppb in Ambazari, Futala, Gorewada and Gandhisagar respectively. The average level
of metals (in ppm) followed the similar order Zn > Cr > Fe > Cd > Pb > Hg > As for both Futala and Gandhisagar
lake respectively. The ranges of variation in present study revealed that the dissolved constitutes of Pb, Cd, Zn, Cr
and Zn was above the ranges of unpolluted water indicating their contamination in water. Cadmium was detected in
traces and ranged from 0.010 mgL-1
to 0.012 mgL-1
, 0.004 mgL-1
to 0.008 mgL-1
, 0.003 mgL-1
to 0.016 mgL-1
and 0.001 mgL-1
to 0.002 mgL-1
in Futala, Gandhisagar, Ambazari and Gorewada lake respectively. Cadmium is
a non essential metal that is toxic even when present in very low concentration. The toxic effect of cadmium is
exacerbated by the fact that it has an extremely long biological half – life and is therefore retained for long periods
of time in organisms after bioaccumulation. Cadmium is a respiratory poison and may contribute to high blood
pressure and heart diseases[21]. Cadmium has been found to be toxic to fish and other aquatic organisms. Its effect
on man includes kidney damage and serves pain in bones (itai – itai in Japan). The level of lead (Pb) was higher
during summer and rainy season as compared to autumn season in all studied lakes. Variations in lead (Pb) level
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Vol.2, No.1, 34-39 (2011)
P.J.Puri et al.
36
varied from 0.014 mgL-1
to 0.026 mgL-1
, 0.018 mgL-1
to 0.022 mgL-1
, 0.025 mgL-1
to 0.034 mgL-1
and 0.011
mgL-1
to 0.021 mgL-1
in Futala, Gandhisagar, Ambazari and Gorewada lake respectively. Adverse Chronic effects
may occur at 0.5 mgL-1
to 1.0 mgL-1
(Pb). At levels greater than 0.1 mgL-1
possible neurological damage in
fetuses and children may possible. The possible source of Pb in studied lakes could be from domestic sewage,
immersion of idols of God and Goddess during festival season and effluent discharge from waste disposal sites as
well as geology of catchments. Chromium was detected with lower levels in autumn and higher throughout
monitoring period during summer and rainy season respectively. The heavy metals concentration in all studied lakes
showed distinct temporal and spatial variations. Among metals, concentration of arsenic (As) remained always
below the detection level throughout study period in entire stretch except Futala and Ambazari lake. Thus it could be
presumed that levels of (As) arsenic in Gorewada lake remained almost in natural level and there was probably no
anthropogenic input in Gorewada lake for its enrichment. Since pH of studied lake water lies in the range of neutral
to alkaline the levels of studied metals could not rise so much as there are natural mechanism to remove these metals
from aqueous solution and prevent from enrichment. A high degree of yearly variation was observed in zinc
concentration. Its yearly variation showed an upward trend. In natural water system Zn remains as either hydroxide
or carbonate form with having almost same solubility which is higher than solubility of existing forms of other
metals. This could be the reason for comparatively higher values of Zn in studied lake water. The average level of
metals followed the order Zn > Cr > Fe > Cd > Pb > Hg > As for both Futala and Gorewada lake respectively.
Higher values of metals in all studied lakes are due to washing activities, recreational activities, immersion of idols
of God & Goddess during and after festival seasons, vehicle washing, farming (agricultural) activities, weathering of
minerals and soils, atmospheric deposition, storm water run off resulting from rainfall, and (poor) sewage. Thus,
water system with enriched toxic metals can serve as reservoirs and may becomes a potential source to supply toxic
metals in the environment. chromium content in studied lakes ranges from 0.028 mgL-1
to 0.042 mgL-1
, 0.028
mgL-1
to 0.036 mgL-1
, 0.014 mgL-1
to 0.028 mgL-1
and 0.016 mgL-1
to 0.018 mgL-1
in Futala, Gandhisagar,
Ambazari and Gorewada lake respectively. Chromium ingestion over admissible limits leads to allergic phenomena
and lung cancer. Mercury is considered to be the most toxic metal. In organic form it enters the human through fish.
Fishes being one of main aquatic organism in food chain may often accumulate large amount of certain metals[22].
Highly significant difference was noticed in case of mercury (Hg) in water samples collected from Futala, Ambazari
and Gandhisagar lake. The concentration of mercury in water varied from 0.005 mgL-1
to 0.018 mgL-1
, 0.008
mgL-1
to 0.018 mgL-1
, 0.010 mgL-1
to 0.021 mgL-1
and 0.001 mgL-1
to 0.003 mgL-1
in Futala, Gandhisagar,
Ambazari and Gorewada lake respectively. The continuous increase in heavy metal contamination in studied lake is
a cause of concern as these metals have ability to bioaccumulate in tissues of various biota’s and may also affect
distribution and density of benthic organisms as well as composition and diversity of faunal communities. Since pH
and temperature affects solubility and toxicity of metals in aquatic ecosystem, this pH and temperature ranges were
also used to access the metal toxicities in studied lakes. Metals such as Cd, Pb and Zn are most likely to have
increased detrimental environmental effects as a result of lowered pH..
Table-1: Heavy metal content (ppm) during different seasons of the year at Futala lake.
Heavy Metals (mgL-1
) Season
Zn Cr Fe Cd Pb As Hg
Summer 0.048 0.042 0.035 0.012 0.026 BDL 0.018
Winter 0.025 0.039 0.022 0.010 0.014 BDL 0.005
Rainy 0.036 0.028 0.024 0.011 0.021 BDL 0.016
BDL :Below Detectable Limit
Table-2: Heavy metal content (ppm) during different seasons of the year at Gandhisagar lake.
Heavy Metals (mgL-1
) Season
Zn Cr Fe Cd Pb As Hg
Summer 0.041 0.036 0.031 0.008 0.022 BDL 0.012
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Vol.2, No.1, 34-39 (2011)
P.J.Puri et al.
37
Winter 0.018 0.028 0.014 0.004 0.018 BDL 0.008
Rainy 0.016 0.030 0.018 0.006 0.020 BDL 0.018
BDL :Below Detectable Limit
Table-3: Heavy metal content (ppm) during different seasons of the year at Ambazari lake.
Heavy Metals (mgL-1
) Season
Zn Cr Fe Cd Pb As Hg
Summer 0.032 0.028 0.025 0.005 0.031 BDL 0.016
Winter 0.025 0.016 0.029 0.003 0.025 BDL 0.010
Rainy 0.021 0.014 0.031 0.016 0.034 BDL 0.021
BDL :Below Detectable Limit
Table-4: Heavy metal content (ppm) during different seasons of the year at Gorewada lake.
Heavy Metals (mgL-1
) Season
Zn Cr Fe Cd Pb As Hg
Summer 0.018 0.020 0.011 0.002 0.016 BDL 0.001
Winter 0.012 0.018 0.012 0.001 0.014 BDL 0.001
Rainy 0.020 0.013 0.001 0.010 0.002 BDL 0.003
BDL :Below Detectable Limit
Fig-1: Heavy metal content (ppm) during different seasons of the year at Futala lake.
0
0.02
0.04
0.06
Zn Cr Fe Cd Pb Hg As
Co
nce
ntr
ati
on
(p
pm
)
Heavy Metals
Summer
Winter
Rainy
Fig.-2: Heavy metal content (ppm) during different seasons of the year at Gandhisagar lake.
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Vol.2, No.1, 34-39 (2011)
P.J.Puri et al.
38
Fig.-3: Heavy metal content (ppm) during different seasons of the year at Ambazari lake.
Fig.-4: Heavy metal content (ppm) during different seasons of the year at Gorewada lake.
CONCLUSION The heavy metal concentration in studied lake showed distinct temporal and spatial variations. There was significant
seasonal variation in metal concentration within the study period. The dry season registered elevated levels of metals
as compared to wet season. Dilution effect of rainy season due to storm run off into receiving lakes and excessive
evaporation of surface water with its attendant pre-concentration of most of metals may be responsible for
observed trend. The results of study have indicated gross pollution of lakes especially regards heavy metals. This
poses a healthy risk to several communities in catchment who rely on these lakes primarily for their domestic
sources without treatment. An elevated level of heavy metals in water is a good indication of man – induced
pollution as a result of (poor) sewage, domestic waste and immersion of idols of God and Goddess during and after
festival season into studied lakes. There were definite upward yearly trends in the concentration of chromium, iron,
lead and zinc in studied lakes which indicated increased input of their pollution load. The levels of mercury (Hg)
and arsenic (As) in studied lake water were comparatively lower. The average level of metals followed the order Zn
> Cr > Fe > Cd > Pb > Hg > As and for all studied lakes. Through some of detected heavy metals are beneficial for
human and plants up to a certain limit; it may be harmful beyond that. Adoption of adequate measures to remove
heavy metals load from industrial waste water, prevention of immersion of idols of God and Goddess along with
fruits, flowers and worship materials along with washing activities are suggested to avoid further deterioration of
lake water quality.
ACKNOWLEDGMENTS The authors hereby acknowledge the kind and wholehearted support of the Dr. S. B. Gholse Director, LIT, RTM,
Nagpur University, Nagpur.
REFERENCE 1. Nagajyoti, P.C., Dinakar, N., Prasad, T.N.V.K.V., Journal of Applied Sciences Research, 4(1) (2008) 110.
2. Friedman, M., Environ. Sci. Technol., 6 (5) (1972) 457.
3. Akeson, M. and D. Munns, Journal Plant Nutri., 13 (1990) 467.
4. Anderson, A. and K. O. Nilsson, Sewish Jounal Agricult. Res., 6 (1976) 15.
5. Antonovics, J., A. D. Bradshw and B.G. Turner : Adv. Ecol. Res., 7 (1975) 1.
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Vol.2, No.1, 34-39 (2011)
P.J.Puri et al.
39
6. Asami Teuro: Sci. Res. Fac. Agric,. Iberaki Univ ., 23 (1975) 43.
7. Azad, A.S., B.R. Arora, B. Singh and G.S. Sekhon: Ind. J. Ecol., II (1) (1984) 1.
8. Borovik, A.S., CRC Press Boca Raton. Florida, 3-5 (1990).
9. Bowen, J. E. : Plant Physiol, 44 (1973) 255.
10. Dutta, I. and Mookerjee, A., Indian J. Environ. Health, 22(3) (1980) 220.
11. Fytianos, K., V. Samanidou and T. Agelidis, Ambia 15 (1) (1986) 42.
12. Mc. Calla, T.M., J.R. Perterson and C.I., Lue ; Ed. Elliot, L.F. and F.J. Stevensn, Sashngton, 28-36 (1977).
13. Vander Werff, M. and J. P. Margaret : Chemosphere. 11(8). (1982).
14. APHA; Standards Methods for the examination of water and waste water, 18th
edition, (1998).
15. Wiklander, L. and Kantol Vahtras : Geoderma 19 (2) (1977) 123.
16. Radhika, C. G. Mini, J., & Gangaderr, T.: Pollution Research, 23, (2004) 49.
17. King, D.L., Journal for the Water Pollution Control Federation , 42(1970) 2035.
18. Khan, M.A. G., & Choudhary, S.H., Tropical Ecology, 35, (1994) 35.
19. Patil, P.R., Chaudhari, D.N. & Kinage, M.S. Environemental Ecology, 22, (2004) 65.
20. Bhatt, L.R. Lacoul, P., Lekhak, H.D., & Jha, P.K. : Pollution Research, 18 (1999) 353.
21. Friberg, L. and C. J. Elinder; Health and Diseases, 18 (1988) 559.
22. Purandara, B.K., Vararajan, N. and Jayashree K. Poll., Res, 22 (2) (2003) 189.
Fig. -5: Map showing Gandhisagar, Ambazari, Gorewada and Futala Lake, Nagpur (MS) India.
[IJCEPR-142/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 40-43
January-April, 2011
Lalita Arora and Sunita Agarwal
Knowledge, Attitude and Practices regarding Waste Management in Selected
Hostel Students of University of Rajasthan, Jaipur
Lalita Arora* and Sunita Agarwal Department of Home Science, University of Rajasthan, Jaipur, India
*E mail: [email protected] Article History:
Received: 7 February 2011
Accepted: 27 February 2011
ABSTRACT
The risk of unhealthy disposal of solid waste is one of the important problems in many societies. Environmental knowledge
attitude practices of young people (like students) appears to be crucial as their point of view ultimately plays an important role in
providing solution to future environmental problems. The study was conducted aiming to find knowledge attitude and practices
of University students with respect to waste management. Total 300 students were included in this study. Data collected by self
administered questionnaire and analyzed, using‘t’ test. It was found that knowledge attitude and practices of University students
regarding waste management was low, less favorable and moderate respectively and correlation between knowledge and attitude,
attitude and practices was not found, but significant correlation was found between knowledge and practices.
Keywords: Knowledge, attitude, practices, waste. ©2011 ijCEPr. All rights reserved
INTRODUCTION With the development of civilization and globalization drastic changes have come in our life style and in every
activity like education, recreation, traveling, feeding, clothing and housing, we are generating lots of wastes. The
modern 'culture of consumerism' has aggravated the waste problem. To this has added the culture of 'disposable'
where large number of goods in the society is being manufactured for 'one time use' and to be discarded as waste
after use. These wastes products create particularly serious problems for the municipalities and its safe disposal is
becoming a serious environmental problem and an ecological crisis is slowly brewing up, threatening to choke the
earth and its life supporting systems.
A number of studies have been carried out by various organizations which provide an estimate about the quantity of
waste generation in various cities. According to Asokan et al [1] about 960 million tones of solid waste is being
generated in India annually, as byproducts during industrial mining municipal, agricultural and other processes. Of
this 350 million tones are organic waste from agricultural sources, 290 million tones are inorganic waste of
industrial and mining sectors and 4.5 million tones are hazardous in nature.
Unhealthy disposal of solid waste is considered as one of the most important problems in many societies. The
problem of waste management has arisen recently in developing countries where there is little history of the
implementation of formal and informal community environmental education awareness program.
Environmental attitude of young people appears to be crucial as they ultimately play a direct role in providing
knowledge based solutions to in coming environmental problems[3,5].
The few studies conducted regarding children and young people, show that the level of environmental awareness is
relatively low [6]. The information acquired is mostly factual in nature and is not systematized.
Begum, R. et al [2] found that majority of the doctors, nurses, and housekeepers have unsatisfactory knowledge and
inadequate practice related to health care waste management.
Keeping all this in view, the present study was planned to analyze their knowledge attitude and practices of hostel
students regarding waste management.
MATERIALS AND METHODS This study attempts to identify the knowledge attitude and practices of University hostel students regarding waste
management. Total 807 students were residing in selected girl’s hostels out of which 300 students were selected for
study. The selection of the respondents was done by stratified sampling method. Students were divided into strata
according to their level of education (PG and UG). 150 students from UG hostels and 150 students from PG hostels
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Vol.2, No.1, 40-43 (2011)
Lalita Arora and Sunita Agarwal
41
were selected by selecting every 3rd number from the list of students. Then each of these strata was further
subdivided according to their stream of education viz Sc and NonSc and 75 students from each stratum were
selected. The self administered questionnaires were used to identify knowledge attitude and practices of University
students in the study area. Before it was used, the questionnaire was pretested in the pilot study. Split half method
was used to calculate the reliability. Reliability of the questionnaire was .96, .94 and .96 for knowledge attitude and
practices questionnaire respectively. Information collected through questionnaire included (1) General information
on respondents including age, education, family type and size etc. (2) knowledge regarding waste management (3)
attitude regarding waste management (4) practices regarding waste management.
The respondents were well informed about the purpose of the study and about the questionnaire by the research
investigator prior to data collection. After collecting data, data were edited and tabulated before data analysis.
Descriptive statistics i.e. percentage, mean and standard deviation were used to describe studied variables. ‘t’ test
and correlation tests were used according to the objective of this study.
RESULTS AND DISCUSSION Knowledge regarding waste management
Knowledge about waste management was enquired using (300) questionnaire. The responses were given scores and
thus the students were categorized as possessing low, medium and high level of knowledge. It was found that
162(54%) of the respondents could be classified as possessing low knowledge, whilst 138(46%) students were
having medium level of knowledge regarding waste management (Table-1).
Attitude regarding waste management
The responses on attitude were classified into less favorable, favorable and most favorable. It was highly striking to
note that majority of hostel students (64.33%) had less favourable attitude towards waste management and only
6.10% (Table-1) were found to have most favourable attitude.
Practices regarding waste management
The responses to practices by respondents are shown in Table-1. Those who had good practices were assumed to be
managing the waste in proper manner and be able protect themselves and environment from negative impact of
waste. From the results of this study it was found that only 1.33% of the respondents could be classified as having
good practices, whilst more than half of the respondents had moderate practices and nearly half of the respondents
140 (46.66%) were found to have poor practices towards waste management. This indicates that they need to
improve their practices regarding waste management.
Hebel-Ulrich, Maja[8] has found that many responses regarding knowledge indicate that the awareness about
hygiene exists, but is not being practiced. Also the observation of several risk behaviors, such as open defecation,
lack of personal hygiene and irresponsible waste management suggests the need for hygiene educational program.
Factors influencing knowledge of the respondents
Factors influencing included in this part of the study were level of education and stream of education. ’t’ test was
used to find out the difference in knowledge scores according to their level of education and stream of education. It
can be observed from the Table-2 that there was a significant difference in the knowledge regarding waste
management base on educational levels of the respondents. It means that PG students have higher scores of
knowledge as compared to UG students. Saini, S. et al [11] measured the knowledge regarding biomedical waste
management. Their results show that consultants, residents, and scientists respectively have 85%, 81%, and 86%
knowledge about the bio medical waste management. Nurses and sanitary staff, operation theatre and laboratory
staff have respectively 60%, 14%, 14%, and 12% awareness of the subject. This shows that the people with higher
education have more awareness about the waste management issues. A significant difference was also observed
between Sc and NonSc students which signify that stream of education makes an impact on knowledge regarding
waste management. According to Ehrampoush, M.H. et.al. [4] the knowledge of the students regarding waste
management was not appropriate. About 66% of students did not participate in segregation and recycling of solid
waste.
Factors influencing attitude of the respondents It was found that level of education did not make any impact on attitude of the respondents regarding waste
management as no significant difference was observed between UG and PG students regarding attitude as shown in
Table-3. Paengkaew, W. et.al [9] observed that majority of Asian students appeared to have lack of environmental
consciousness and attitude needed to protect their environment. Therefore it is important to develop skills,
awareness, and attitude and put in to practice.
But stream of education is showing a significant difference on attitude. This may be due to that Sc students have
some chapters on environmental pollution and waste management in their course and therefore they are little aware
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Vol.2, No.1, 40-43 (2011)
Lalita Arora and Sunita Agarwal
42
regarding waste management. As per the study done by Saini, S. et al [11] measured the attitude regarding
biomedical waste management of doctors, nurses, and other support staff. They found that the people with higher
education and knowledge have better attitudes towards the subject.
Factors influencing practices of the respondents
Practices of students were affected by both the variables i.e. level of education and stream of education. To find out
the difference both the variables ‘t’ test was performed and ‘t’ values were found to be 3.86 and 4.14 (Table-4) for
level of education and stream of education respectively. From ‘t’ values a significant difference between UG V/s PG
and between Sc V/s NonSc was found suggesting that level and stream of the respondents affect the practices
regarding waste management. Pothimamaka, J. (2008) found that more than half of the house holds had no waste
separation practices and they concluded that their practices were not appropriate towards solid waste management
and people must be taught to deal with solid waste by separating it in their homes, schools and work places.
Association between knowledge, attitude and practices regarding waste management.
Pearson ‘r’ correlation test was used to find out the association between knowledge, attitude and practices regarding
waste management. As shown in Table-5 it was observed that there was a significant association between
knowledge and practices with the correlation coefficient of 0.167 at 0.01 levels. It means those who possess good
knowledge also have good level of practices, thus are able to manage the waste in proper manner.
Grodzinska Jurczak,M.S and Friedlin, K. [6] also found that a correlation between the level of students’ knowledge
and their activities was found regarding waste management.
According to the Table-5 no significant association between knowledge and attitude with correlation coefficient of
0.04 and attitude and practices with correlation coefficient of 0.003 was found for waste management. Same results
were found from Wai, S. Tantrakarnapa, K and Huangprasert, S. [12] that there was a significant association
between knowledge and practices with correlation coefficient of 0.39 and knowledge and attitude with correlation
coefficient of 0.289. But there was no significant association between attitude and practices for environmental
sanitation.
CONCLUSION The majority of the respondents have unsatisfactory knowledge attitude and inadequate practices related to waste
management. This study has shown a need to improve the knowledge about waste management to protect
environment from negative impact of waste. It is recommended to implement the need based training programme for
students at their school hostels and work places.
Table-1: Knowledge Attitude and Practices regarding waste management
S.No. variables Category Number (%)
Low 162 54
Medium 138 46
1.
Knowledge
High - -
Less favourable 193 64.33
Favourable 89 29.66
2.
Attitude
Most favourable 18 6.10
Poor 140 46.66
Moderate 156 52
3.
Practices
Good 4 1.33
Table-2: Factors influencing Knowledge of the respondents
S.No. Factors Mean S.D. ‘t’ value
1. Level of education
UG 10.21 2.73
PG 10.87 2.95
2.00*
2. Stream of education
Sc. 11.09 2.86
NonSc 9.99 2.76
3.38*
* Significant ** Non Significant
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Vol.2, No.1, 40-43 (2011)
Lalita Arora and Sunita Agarwal
43
Table-3: Factors influencing Attitude of the respondents
S.No. Factors Mean S.D. ‘t’ value
1. Level of education
UG 62.61 22.02
PG 66.31 23.44
1.40**
2. Stream of education
Sc. 67.17 24.15
NonSc 61.75 21.05
2.06*
* Significant ** NonSignificant
Table-4: Factors influencing Practices of the respondents
S.No. Factors Mean S.D. ‘t’ value
1. Level of education
UG 20.09 8.00
PG 23.18 5.03
3.86*
2. Stream of education
Sc. 23.29 7.38
NonSc 19.99 6.37
4.14*
* Significant ** NonSignificant
Table-5: Association between variables
S.No. Variables Co-efficient of reliability
1. Knowledge and Practices 0.16*
2. Knowledge and Attitude 0.04**
3. Attitude and practices -.003**
* Significant ** Nonsignificant
REFERENCES 1. Asokan, P., Sexena, M. and Asolekar, S., Building and Environment., 42 (2007) 2311.
2. Begum. Ara, Rawshan and Pereira, Jacqueline, Joy., Asian Journal of Water, Environment and Pollution.,
5(3) (2008) 15.
3. Bradley, C. J., Waliczek. T. M. and Zajicek, J. M., Journal of Environmental Education., 30(3) (1999) 17.
4. Ehrampoush, M.H., Baghiani Moghadam, M.H., Iranian Journal of Environmental Health Science
Engineering., 2(2) (2005) 26.
5. Eagles, P.F.J. and Demare, R., Journal of Environmental Education., 30(4) (1999) 33.
6. Grodzinska-Jurczak, M.S., Resource Conversion and Recycling., 32(2) (2001) 85.
7. Grodzinska-Jurczak, M.S. and Friedlein, K., Environmental Science and Pollution Res., 3(3) (2002) 215.
8. Hebel-Ulrich, Maja., Danish Committee for Aid to Afghan Refugees (DACAAR) (2005) www.dacaar.org.
9. Paengkaew, W., Roongtawanreongsri, S. and Kittitoronkool, J., Paper presented at SEAGA Conference.,
28-30 November (2006) Singapore.
10. Pothimamaka,J.,Environment Asia., (Available online at www.tshe.org/EA) (2008) 43
11. Saini, S., Nagarajan, S.S. and Sharma, R. A., Journal of the Academy of the Hospital Administration.,
17(2) (2005).
12. Wai,S.,Tantrakarnapa, K. and Huangprasert, S., Thai environmental Engineering Journal., 19(2) (2005) 19.
[IJCEPR-149/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical RePharmaceutical RePharmaceutical RePharmaceutical Researchsearchsearchsearch
Vol. 2, No.1, 44-48
January-April, 2011
Yogesh B. Patil
Utilisation of Thiocyanate (SCN-) by a Metabolically Active Bacterial
Consortium as the Sole Source of Cellular Nitrogen
Yogesh B. Patil Department of Energy and Environment, Symbiosis Institute of International Business (SIIB)
[A Constituent of Symbiosis International Institute (SIU)], G. No. 174/1, Rajiv Gandhi Infotech Park,
Hinjewadi, Pune – 411 057, Maharashtra, India.
E-mail: [email protected] Article History:
Received:26 April 2011
Accepted:28 April 2011
ABSTRACT Thiocyanate (SCN-), a toxic chemical species from cyanide family, consists of both carbon (C) and nitrogen (N) in equimolar
ratio and is being emanated through liquid effluents by several industrial processes. Since bioremediation technologies for waste
treatment are gaining enormous importance in the recent times; microbial treatment of SCN- is therefore being researched
worldwide. However, utilisation of SCN- by microbes as a suitable growth substrate (C and/or N source) is poorly understood
and presents the problem in waste treatment systems. A heterotrophic bacterial consortium comprising of three Pseudomonas sp.
isolated from activated sludge and having potentials for environmental clean-up, was capable of utilising thiocyanate (SCN-)
from aqueous solutions as the sole source of cellular nitrogen (N) in the presence of carbon (C) source viz. glucose. The
consortium ceased to grow and degrade SCN- when supplemented with C and N sources alone. Diauxie pattern was observed
when the consortium culture was supplied with two N sources (NH4Cl and SCN-) in the presence of glucose as a source of carbon
(C). NH4Cl was the preferred growth substrate utilised by the bacterial consortium followed by SCN-.
Keywords: Biodegradation, Bacterial consortium, Diauxie, Nitrogen source, Thiocyanate. ©2011 ijCEPr. All rights reserved
INTRODUCTION Industrial processes like metal extraction, dyeing, photo-finishing, thiourea, pesticide production and electroplating
industries produce large quantities of thiocyanate (SCN-) bearing effluents [6]. The concentration of SCN
- in these
effluents is in the range of 5 to 100 mg/L. Since SCN- is toxic to all living cells [14], it is imperative for the
industries to detoxify the effluents prior to their discharge in environment. Several physical-chemical technologies
have been reported for the SCN- removal [4]; the most widely being used is chlorination. However, conventional
methods are beset with problems that are environmentally hazardous and fail to bring SCN- level within safe limits.
Moreover, SCN- content in the waste inhibits the degradation of other pollutants (like cyanide and metal-cyanides)
present in the waste, and therefore, has detrimental impact on aquatic flora and fauna. It is, thus, necessary to
develop an alternative treatment process capable of achieving high degradation efficiency at low-cost.
Bioremediation technologies using microorganism for detoxification of waste chemical is an eco-friendly
alternative. Considerable literature is available on the removal of toxic C-1 compounds like free cyanide and metal
cyanides by metabolically active [5,9] and metabolically inactive (passive) microorganisms [8]. In the recent times
Thakur and Patil (2009) had reported the removal of SCN- from solutions using low-cost waste biomass [13].
Although several reports are available on microbial SCN- degradation [4,11,12], utilisation of it by microbes as a
suitable growth substrate (C and/or N source) is poorly understood. Lack of scientific knowledge in this regard may
pose problems in the biological treatment systems. The present paper highlights some key laboratory experiments
that confirm the degradation of SCN- by a heterotrophic bacterial consortium utilising it as the sole source of cellular
N in the presence of external carbon (C).
MATERIALS AND METHODS A heterotrophic bacterial consortium comprising of three Pseudomonas species and capable of utilising SCN
- was
isolated from activated sludge by an enrichment culture technique [11]. The consortium was grown for 24 - 48 h in
M-9 minimal salts medium – MSM [7] with 1 ml/L micronutrient solution [3], which contained SCN- (50 mg/L ≅ 1
mM) and glucose (10 mM) as the sole N and C & energy sources, respectively. The medium was totally free of
synthetic organics like peptone, beef extract and yeast extract. Bacterial cell suspension (0.1 ml) containing 108
cells/ml was used as an inoculum. Batch culture experiments on SCN- biodegradation were performed under aseptic
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Vol.2, No.1, 44-48 (2011)
Yogesh B. Patil 45
and aerated conditions in 250 ml Erlenmeyer flasks with 100 ml M-9 MSM. The medium was supplemented with C
and N sources with various permutations and combinations as shown in Table 1.
Table-1: Experimental manipulation of carbon and nitrogen sources in various proportions for bacterial growth
The pH of medium before starting the experiment was adjusted to 7.0. All the flasks were incubated at 30°C in a
rotary shaker incubator (Remi, CIS-24 BL) at 150 rpm for 48-72 h. Suitable controls were run simultaneously along
with experimental flasks to detect air stripping or auto-oxidation of thiocyanate, if any. Experiments were repeated
twice to confirm the results. Analytical grade chemicals were used in all the experiments. Reagents were prepared in
RO (Sartorius, Arium-61315) water (conductivity <5 µS) and refrigerated (4°C). SCN- content was measured
spectrophotometrically (Spectronic-20D) as per the Standard Methods [1]. pH was determined using pH meter
(Elico, Ll-120). Bacterial population was checked microscopically (Metzer, 778A) using Neubauer’s chamber (Fein-
Optik, Blankenburg) and by total viable count (TVC) procedure.
RESULTS AND DISCUSSION The data in Fig. 1 shows the growth of bacterial consortium when SCN
- was supplemented in the medium as both C
and N source. The results clearly indicated that the consortium failed to utilise SCN- as either C or N source. The
SCN- concentration and bacterial population remained constant throughout the experimental period of 45 h. The data
in Fig. 2 depicts that the bacterial consortium was capable of utilising SCN- (with an efficiency of > 99%) as the sole
source of cellular N in the presence external C source like glucose, wherein increase in bacterial cell density from
105 to >10
8 cells/ml was observed with simultaneous decrease in SCN
- level from 50 to < 0.1 mg/L. In uninoculated
control, the SCN- level remained unaltered (Fig. 2). SCN
- when supplemented with NH4Cl, inhibited growth of
bacterial consortium (Fig. 3). Fig. 4 shows diauxic growth (diauxie) pattern of the bacterial consortium when SCN-
was supplied along with C and N sources (Glucose and NH4Cl). It was observed that when SCN- and NH4Cl were
supplied, the consortium did not utilise both N sources simultaneously. The consortium initially utilised NH4Cl and
then SCN- as N source. The growth of consortium in the first 25-30 h and the unchanged levels of SCN
- in the same
period confirmed utilisation of NH4Cl during first phase and later the SCN-. This resulted in a biphasic (two phase)
growth pattern known as diauxie / diauxic growth (Fig. 4).
The prime objective of the present work was to elucidate the potentials of isolated bacterial consortium for its
utilisation of SCN- from aqueous solution as (i) both C and N source or (ii) only C source or (iii) only N source for
growth. Revealing this fact is important in biological treatment of wastewater because if toxic chemical like SCN- is
used by the consortium as both C and N source, then at practical scale, external supplementation of nutrients
wouldn’t be required, which in author’s opinion, would be beneficial from the economic point of view to the user
industries. This holds true even for the wastewaters containing other toxic compounds like cyanide and metal-
cyanides. However, in the present study, the bacterial consortium failed to utilise SCN- as both C and N source (C/N
molar ratio=1). This fact clearly indicates that toxic SCN- compound presents problem to the consortium for growth
utilising it as suitable substrate (Fig. 1). Thus, a sufficiently high concentration of SCN- to support appreciable
growth might prove to be too toxic to allow growth to occur. Since the concentration of N required for a given
amount of growth is less than the requirement for C, it might be easier for bacterial consortium to utilise SCN- as the
source of N in the presence of a separate source of C and energy. Therefore, microorganisms capable of degrading
SCN- as the source of N were isolated by an enrichment technique [11]. The consortium in the present study clearly
showed (Fig. 2) the utilisation of SCN- as the sole N source in the presence of external C source like glucose (C/N
molar ratio=11). Therefore, from the process development point of view, it is essential to supplement some cheaper
Combinations Carbon Source Nitrogen Source Overall C/N Molar Ratio
in M-9 MSM
A Potassium thiocyanate i.e.
KSCN (50 mg/L ≅ 1 mM)
KSCN (50 mg/L) 1
B Glucose (10 mM) KSCN (50 mg/L) 11
C KSCN (50 mg/L) Ammonium chloride i.e.
NH4Cl (1 mM)
0.5
D Glucose (10 mM) KSCN (50 mg/L) +
NH4Cl (1 mM)
5.5
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Vol.2, No.1, 44-48 (2011)
Yogesh B. Patil 46
source of C like molasses, which is readily available in developing country like India at cheaper rate. In the earlier
studies, Patil (1999) had successfully demonstrated the use of molasses as the source of C to develop a microbial
technology for metal cyanides biodegradation/removal from wastes utilising it as the sole source of N [10].
Uninoculated controls run simultaneously along with the experiments did not show any decrease in SCN- levels
(Fig. 2) confirmed that biodegradation of SCN- was the predominant reaction taking place by the bacterial
consortium. There are a few reports, which describe microbial SCN- degradation utilising it as the sole N source [4,
12]. When SCN- was supplied as the sole C source in the presence of external N (viz. NH4Cl), the consortium ceased
to grow keeping the 50 mg/L of SCN- amount unaltered (Fig. 3). This might be due to the higher amount of
available N (2 mM) compared to the C (1 mM) source (C/N molar ratio=0.5). Obviously, the consortium culture will
find it more difficult to obtain the energy from low amount of C and utilise the toxic SCN-. Diauxic (Biphasic)
growth pattern was observed (Fig. 4) when two N salts (i.e. SCN- and NH4Cl) along with one C source (glucose)
were supplied to the consortium. NH4Cl was the preferred growth substrate utilised by the bacterial consortium
followed by SCN- degradation suggests that SCN
- utilisation by consortium is inducible. Diauxie pattern in
Escherichia coli (and many other microorganisms) in the presence of two C sources viz. glucose and lactose is a
well known example [2]. However, there are no reports on the diauxie pattern when two N sources like SCN- (a
toxic compound) and NH4Cl are supplemented and therefore, this paper may be considered as the first report. In the
present study, the diauxic growth experiments (Fig. 4) showed rapid removal/decrease of toxic SCN- from the
medium within 25 h. This was because in the first phase of diauxie, the consortium grew to a substantial level (cell
density from initial 105 to final >10
8 cells/ml) utilising glucose and NH4Cl. After the exhaustion of NH4Cl from the
medium, the consortium in the second phase (after a lag of 10-12 h) degraded toxic SCN- rapidly utilising it as the
sole N source in the presence of glucose (10 mM) as C. It has not escaped through author’s notice that high initial
cell density leading to rapid SCN- biodegradation in diauxie experiment, immediately suggests its possible
application in wastewater treatment reactor vessels capable of retaining high microbial biomass by way of
immobilisation, which is the key to hasten biodegradation of SCN-.
0
10
20
30
40
50
60
0 10 20 30 40
Time (h)
Th
ioc
ya
na
te C
on
c.
(mg
/L)
4.5
5
5.5
6
6.5
7
7.5
8
8.5
9
Lo
g (
No
. o
f b
ac
teri
al c
ells
)
Fig.-1: Supplementation of SCN
- as both C and N source. Cessation of bacterial growth (�) and unaltered SCN
-
concentration (▲)
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Vol.2, No.1, 44-48 (2011)
Yogesh B. Patil 47
0
10
20
30
40
50
60
0 10 20 30 40
Time (h)
Th
ioc
ya
na
te C
on
c. (m
g/L
)
4.5
5
5.5
6
6.5
7
7.5
8
8.5
9
9.5
Lo
g (
No
. o
f b
ac
teri
al c
ells
)
Fig.-2: Utilisation/Degradation of SCN
- by bacterial consortium as the sole source of N in presence of external C
source. Growth of bacterial consortium (▲) with simultaneous decrease in SCN- concentration (●); SCN
-
concentration in the absence of consortium (�)
0
10
20
30
40
50
60
0 10 20 30 40
Time (h)
Thio
cyanate
Conc. (m
g/L
)
4.5
5
5.5
6
6.5
7
7.5
8
8.5
9
Log (N
o. of bacte
rial cells)
Fig.-3: SCN
- as the sole C source with external supplementation of N (NH4Cl). Growth of bacterial consortium (▲);
SCN- concentration in presence (●) and absence of consortium (�)
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Vol.2, No.1, 44-48 (2011)
Yogesh B. Patil 48
0
10
20
30
40
50
60
0 10 20 30 40 50 60 70
Time (h)
Thio
cyanate
Conc. (m
g/L
)
4.5
5.5
6.5
7.5
8.5
9.5
10.5
11.5
12.5
Log (N
o. of bacte
rail c
ells)
Fig.-4: Diauxic growth pattern exhibited by bacterial consortium in the presence two N sources (SCN
- and NH4Cl)
in presence of glucose as C source. Growth of consortium (�) and SCN- degradation (▲) in the presence of two N
sources; SCN- concentration in absence of consortium (●); Cessation of bacterial growth in absence of either N or C
source (�)
CONCLUSION Foregoing experimental results and discussion conclude that the bacterial consortium isolated from activated sludge
is capable of utilising/degrading SCN- as the sole source of cellular N from aqueous solutions in the presence of
external C source.
ACKNOWLEDGEMENT This research was supported by University Grants Commission (UGC), WRO, Pune through a grant to YBP.
REFERENCES 1. APHA, AWWA, WPCF, Standard Methods for the Examination of Water and Wastewater, Washington, DC,
18th
Edn. (1998).
2. Atlas, R.M., Principles of Microbiology, McGraw-Hill, New York, (1997).
3. Bauchop, T., Elsden, S.R., General Microbiology, 23 (1960) 457.
4. Bipinraj, N.K., Joshi, N.R., Paknikar, K.M. Biohydrometallurgy: A Sustainable Technology in Evolution, In:
International Biohydrometallurgy Symposium, Athens, (2003) 491.
5. Gurbuz, F., Ciftci, H., Akcil, A., Karahan, A.G. Hydrometallurgy, 72 (2004) 167.
6. Hughes, M.N. General Chemistry, In: Newmann, A.A., (Ed.), Academic Press, London (1975).
7. Millar, J.H., Experiments in Molecular Genetics, Cold Spring Harbour, NY: Cold Spring Harbour Laboratory
(1972).
8. Patil, Y.B., Paknikar, K.M., Biotechnology Letters, 21 (1999) 913.
9. Patil, Y.B., Paknikar, K.M., Process Biochemistry, 35 (2000) 1139.
10. Patil, Y.B., Ph.D. Thesis, University of Pune, Pune, India (1999).
11. Patil, Yogesh B., Research Journal of Chemistry and Environment, 12(1) (2008) 69.
12. Sorokin, Dimitry Y., Tourova, Tatyana P., Lysenko, Anatoly M., Kuenan, Gigs J., Applied and Environmental
Microbiology, 67(2) (2001) 528.
13. Thakur, Ravindra Y., Patil, Yogesh B., South Asian Journal of Management Research, 1(2) (2009) 85.
14. Westley, J. Cyanide and Sulfane Sulfur. In: Vennesland, B., (ed.), Academic Press, London (1981) 201.
[IJCEPR-163/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 49-51
January-April, 2011
K.Tamilselvan et al.
Construction of An Open Loop Temperature Control System for Thin Film
Fabrication in PC Based Instrumentation
K.Tamilselvan*,1
, K.Anuradha2, S.Deepa
2 , O.N.Balasundaram
2 and S.Palaniswamy
2
*,1Department of Electronics, PSG College of Arts and Science, Coimbatore-641 014. 2Department of Physics, PSG College of Arts and Science, Coimbatore-641 014.
E-mail: [email protected] Article History:
Received: 4 April2011
Accepted: 27 April 2011
Abstract
The paper describes an open loop control for controlling the chemical hot bath temperature in PC based dip coating system for
thin film fabrication. The control of temperature is achieved by changing the duty cycle and supply voltage. The function of the
duty cycle at each supply voltage in obtaining maximum temperature is studied. The open loop control of temperature is
implemented using PC based instrumentation. The temperature control system is optimized using relation between the duty cycle
and temperature. The performance of this system is compared to that of a conventional close loop controller on a laboratory test.
Results are presented that shows a good control of the hot bath temperature.
Keywords: open loop control, PC based dip control system, duty cycle. ©2011 ijCEPr. All rights reserved
INTRODUCTION The use of personal computers in automation systems is widespread. The realization of PC-based automation
systems has been allowed by several factors, among them are-
1. The impressive grown of the PC’s performances which ensures the possibility of locating and executing,
inside the same machine, both the types of tasks involved: real-time control and monitoring /supervision.
2. The use of high level language allows for an efficient scheduling of the tasks assigning higher priorities to
the most critical ones.
3. Increasing availability of devices equipped with bus interfaces which makes possible, the realization of an
automation system using hardware/software products of different manufacturers.
In the first part of this paper, we describe the general structure of the PC-based dip coating unit with open loop
temperature control and an experimental evaluation of its performance using PWM technique.
MATERIALS AND METHOD A Personal Computer with Pentium processor, 256 MB RAM and 80 GB Hard disk drive along with external driver
circuits are used. A small mechanical arrangement driven by stepping motor is used to move the substrate upward /
downward. The stepper motor drives the screw rod, moving pulley and substrate holder. The proper gear reduction
mechanism and well polished gear teeth wheels are employed to avoid mechanical slip and jerk. The screw rod is
precisely machined to give smooth upward and downward movement. The control signals are taken from printer
port or Programmable Peripheral Interface ( PPI ) card. Transistor driver controls the voltage applied to heater of
hot bath. The programs written in C++ are executed. The instrumental setup is shown in figure.
Fig.-1: Instrumental Setup
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Vol.1, No.2, 49-51 (2011)
K.Tamilselvan et al.
50
100V
20
30
40
50
60
70
80
1 8 15 22 29 36 43 50 57 64 71 78 85 92 99 106 113 120
TIME(S)
TE
MP(C
ELS
IUS)
ON 2s
4s
6s
8s
10s
12s
14s
16s
18s
Graph- i: Variation of temperature with time for 100V
and .05Hz for various duty cycles.
150V
0
10
20
30
40
50
60
70
80
90
100
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64 67 70 73 76 79 82 85 88 91
TIME(s)
TE
MP
(celc
ius)
ON 2s
4s
6s
8s
10s
12s
14s
16s
18s
Graph-ii: Variation of temperature with time for 150V
and .05Hz for various duty cycles.
200V
0
20
40
60
80
100
120
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64 67 70 73 76 79 82
TIME(s)
TE
MP(c
elc
ius)
ON 2s
4s
6s
8s
10s
12s
14s
16s
18s
Graph-iii: Variation of temperature with time for 200V
and .05Hz for various duty cycles.
VOLTAGE VS TEMP
0
20
40
60
80
100
120
100 120 140 160 180 200 220
VOLTAGE(V)
TE
MP
(CE
LS
IUS
)
2S
4S
6S
8S
10S
12S
14S
16S
18S
Graph-iv: Plot with voltage and temperature for various
duty cycles. DUTYCYCLE VS TEMP
0
20
40
60
80
100
120
0 2 4 6 8 10 12 14 16 18 20
DUTYCYCLE(s)
TE
MP
(celc
ius)
100V
150V
200V
Graph-v: Plot with duty cycle and temperature for fixed voltages.
Fig.-3: Different Graphs
RESULTS AND DISCUSSION Temperature control The temperature of the hot bath can be controlled by two methods namely,
i. Closed loop control and
ii. Open loop control
Closed loop control
Closed loop control system requires a sensor, a feedback circuit and a complex controller circuit, which is a
drawback of the closed loop control. On the other hand the temperature control in this system can be done very
precisely with a variation of 1ºC or 2ºC.
Open loop contropl
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Vol.1, No.2, 49-51 (2011)
K.Tamilselvan et al.
51
An open loop control is simple.It does not require a sensor, a feedback circuit and also a complex circuit, which adds
to the advantages of a open loop control. It requires only a driver circuit to control the temperature of the hot bath.
In this system, temperature control can be done by using PWM technique , ie
i. by varying the duty cycle
ii. by varying the frequency
where,
Duty cycle = ON time/ Total Time
when the duty cycle decreases the output voltage also decreases and when the duty cycle increases the output
voltage also increases.
Fig.-2: Driver circuit
Optimization
The temperature of the hot bath varies accordingly with duty cycle, frequency and the supply voltage.The change in
temperature of hot bath for fixed interval of time is studied until the temperature of the hot bath reaches the
saturation temperature for fixed frequency and supply voltage . The change in temperature is plotted, graphically.
For a supply voltage of 100V the saturation temperature is minimum and varies according to the duty cycle.
Similarly for 150V and 200V the saturation temperature varies according to the duty cycle.
From these observations we can fix the required temperature of the hot bath by fixing the corresponding duty cycle
and the voltage.
CONCLUSIONS This newly designed simple instrumental setup is cost effective and convenient for preparation of Thin Films. The
well known technique PWM is used to control the temperature. By changing the ON time and keeping the TOTAL
time unchanged, the temperature can be increased or decreased. The proper mechanical setup and programming
techniques improves the system performance.
REFERENCES 1. Maissel, L.I. , Glang R. , Handbook of Thin Films Technology, McGraw- Hill, New York, 1970.
2. Ohring M., The Materials Science of Thin Solid Films, Academic Press, New York, 1992.
3. George W. Gorsline ,Computer organisation, hardware/software, PHI.
4. Stephen J. Bigelow ,Trouble shooting Maintaining & Repairing PC’s, Tata McGraw Hill Co, New Delhi 1999.
[IJCEPR-159/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 52-55
January-April, 2011
Alok Kr. Singh et al.
Accumulation of Natural Antioxidants in Ferns Exposed to Mutagenic Stress
Alok Kr. Singh1, Santosh Kr. Singh
2*, Satish K. Verma
3, H.V. Singh
1, A.K. Mishra
4, Pavan K.
Agrawal3, Abhishek Mathur
3 and Md. Aslam Siddiqui
5
1Department of Botany, S.G.R. P.G. College, Dobhi, Jaunpur *,2Department of Biotechnology, S.B.S. (P.G.) Institute of Biomedical Sciences and Research,
Balawala, Dehradun, Uttarakhand 3Sai Institute of paramedical and allied Sciences, Dehradun 4Kashi Naresh Rajkiya Snatkottar Mahavidhyala, Gyanpur, Bhadohi 5Baba Farid Institute of Technology, Dehradun.
*E-mail: [email protected] Article History:
Received:3 April 2011
Accepted:10 April 2011
ABSTRACT Different species of ferns were analyzed for the modulations in the pool of non-enzymatic antioxidants in response to the maleic
hydrazide treatments. Treatments at very low doses were found to trigger the accumulation of both ascorbate and proline
contents. Total amount of protein and chlorophyll contents showed varying degree of sensitivity in all cultivars of ferns. Proline
accumulation was found to be high in treated plants compared with control. Proline, ascorbate and flavonoid contents were found
to be accumulated in all plants exposed to high doses of maleic hydrazide. All the three species showed high proneness towards
the mutagen. Improved tolerance in treated plants might be explained on the basis of the elevated level of enzymatic and non-
enzymatic antioxidants.
Key Words: Mutagen, Ascorbate, Proline, protein, Ferns ©2011 ijCEPr. All rights reserved
INTRODUCTION Ferns are found abundantly in many different habitats of the world. They were the dominant part of the vegetation
during the Carboniferous Period which is called as ‘Age of Ferns’. Most of the ferns of the Carboniferous became
extinct but some later evolved into our modern ferns. There are about 12,000 species in the world today [2]. Three
fern species were selected for the present study: Cheilanthes farinose, Lygodium scandens and Adiantum caudatum.
The plants prefer light (sandy), medium (loamy) and heavy (clay) soils [12, 13]. The plants are dominant in soils of
acidic, neutral and alkaline nature.
Plants possess many antioxidants, usually classified in two broader categories. They are: enzymatic antioxidants and
non-enzymatic antioxidants. The alterations in the activities of antioxidants is observed in the plants exposed to
different environmental stresses such as drought, heavy metals, pesticides, ultraviolet radiations etc. Since human
activities are increasing the level of pollutants in the environment day by day, it has become an interesting area of
research to observe their effects on plant communities (Producers of the Ecosystems). The damage to the biological
ecosystems may be measured in terms of the morphological and biochemical alterations in primary producers.
Numerous studies have been conducted on photosynthetic enzymes, pigments, proteins, seed patterns and
antioxidant compound contents in plants. Maleic hydrazide is one of the agents that have been found to bring
heritable alterations in the genes, chemical mutagens are undoubtedly very potent ones which can induce genetic or
physical alterations in dormant seed or spore. This is a strong mutagen that induces mitotic inhibition and
cytological abnormalities in a number of higher plants [4, 8, 16]. It possesses growth regulating properties [19, 22].
It is a known depressor of auxin transport in plant. Stem growth, root growth and seed germination can be regulated
by its treatment.
The presence/absence and increasing/decreasing property of antioxidant compounds, production of free radicals and
the amount of lipid peroxidation in terms of elevated level of antioxidants might provide significant clues to assess
and evaluate the antioxidant potential of various Cheilanthes species against environmental stresses. Plants are able
to develop special mechanisms for adjusting the changed environment. Many groups of stresses like heavy metals,
ultraviolet radiations etc are shown to generate singlet oxygen and other active oxygen species at various sites of
photosynthetic electron transport chain [9] and affect the growth of plant. Many studies have been done with
emphasis on morphological, biochemical and genetic characteristics of Cheilanthes rufa, Lygodium scandens and
Adiantum caudatum with respect to ultraviolet radiations, gamma radiations and various light qualities like red light,
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Vol.1, No.2, 52-55 (2011)
Alok Kr. Singh et al.
53
blue light etc. The present study was done with setting forth the objective of studying the effect of Maleic hydrazide
on the accumulation of non-enzymatic antioxidants in different fern species.
MATERIALS AND METHODS Organisms and culture conditions
Spores of Cheilanthes rufa, Lygodium scandens and Adiantum caudatum were collected from plants growing in the
kushmi forest of Gorakhpur (a tarai area of north India). The spores were surface sterilized with 2% sodium
hypochlorite solution and then sown uniformly on 25 ml of autoclave sterilized (15 1b/in2) inorganic medium at pH
5.4 in petri dishes [20]. Sowing of spores was done in an inoculation chamber fitted with germicidal UV lamp
(USA). Then the plates were maintained at 24 ± 20c under continuous white fluorescent illumination at the intensity
of 2700 lux. Spores for treatments were placed in liquid nutrient media for 72 hours and then subjected to various
concentrations of maleic hydrazide. Each experiment was conducted in triplicates.
Extraction and estimation of total Protein, Malondialdehyde and peroxide contents
Growth determination was done by estimating total protein contents after 12 days of treatments. Protein contents
were determined using Folins-Lowry method with lysozyme as the standard [14]. Total amount of hydrogen
peroxide radicals was estimated by using ferrithiocyanate method as described by Sagisaka (1976) [18].
Standardization of H2O2 was performed to minimize the interference of catalase. The level of lipid peroxidation was
measured in terms of total malondialdehyde (MDA) contents. The reaction reagent consisted of 0.4 N TCA + 19.68
ml of distilled water + 0.4 ml of HCl + 100mg TBA [11]. Prepared leaf extract (in phosphate buffer) was added to
the reaction reagent and absorbance was taken at 532 nm.
Flavonoids: Extraction and Estimation
Flavonoids were extracted in mutagen-treated and untreated fern leaflets by using the method of Mirecki and
Teramura (1984). Extraction mixture consisted of acidified methanol (methanol: water: HCl, 78: 20: 2, v/v) +
leaflets, incubated for for 24h at 40C. The filtered extract was then used for measuring the absorbance at 320 nm,
which is indicative of relative concentration of UVB absorbing pigments [15]. Flavonoid contents were expressed as
absorbance g-1 fresh mass of tissue at 320 nm.
Extraction and estimation of Proline contents
Proline contents in leaf homogenate of mutagen- treated and untreated cultures were estimated according to the
standard method [3]. Proline contents in unknown samples were calculated by comparing with standard curve of L-
proline. Amount of proline is represented in terms of µg g-1
FW.
Ascorbic acid estimation
Ascorbic acid was extracted by dehydrating ascorbic acid by shaking it with acid washed NORIT* in the presence of
acetic acid. After coupling with 2, 4-Dinitrophenyl hydrazine, the solution is treated with sulfuric acid to produce
the red color whose absorbance was measured at 540 nm.
*Acid washed NORIT preparation: 200 gram NORIT (charcoal) is suspended in 1000 ml of 10% HCl, heated upto boiling point and filtered under
suction. The cake is removed and stirred with 1000 ml water and filtered. This procedure is repeated until the
washing give a negative test for Fe3+
ions. The NORIT is then dried overnight at 110-1200C.
RESULTS AND DISCUSSIONS Results observed were found to be variable with different fern species. Overall growth and accumulation of
ascorbate, proline and flavonoid contents showed modulations in vales in response to the mutagenic chemical–
Maleic hydrazide. Growth measured in terms of protein contents showed initial increase at low doses of mutagen (%
control increase = 16-22% in all three ferns). But the high doses were found to reduce the total protein contents
speedily (5-60% at 125 ppm as compared to the control) (Figure 1a). The declining trend in protein contents
continued with rising concentration of maleic hydrazide. Initial recovery in protein contents might be explanined on
the basis of increase in the pool of enzymatic antioxidants that help plants in overcoming the oxidative stress [6, 7]
The present study might give us clues for the impacts of mutagens on total biomass yield.
Cells contain important non-enzymatic antioxidants such as carotenoids, ascorbic acid, proline, glutathione, α-
tocopherol etc., for mitigating the toxic effects of free radicals and AOS (active oxygen species) under oxidative
stress. In the present work, ascorbate contents showed increase in the amounts (% control increase= 10-45% in all
plants) with mutagen exposure up to 16-62 ppm and the decrease at high dose (6-15% after 125 ppm exposure of
maleic hydrazide), compared to untreated samples. (figure 1 b). There are two possibilities regarding increase in
ascorbic acid contents; either its synthesis has increased or its regeneration rate through the Asada-Halliwell
pathway has increased (as observed in Ulva fasciata) [21].
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Vol.1, No.2, 52-55 (2011)
Alok Kr. Singh et al.
54
The chemical evolution and significance of flavonoids has been assumed to play an important role in overcoming
the oxidative stress in cells [17]. Evidences suggest that the presence of flavonoids in UV-B irradiated leaves could
alter the perception or response of other defense mechanisms. Presently, flavonoid contents showed enhanced
synthesis in maleic hydrazide treated fern species- C. rufa, Lygodium scandens and Adiantum caudatum. A %
control increase of 5-40% was observed at the doses of 16-62 ppm but at the high dose (125 ppm), the values
decreased to 25%, 20% and 30% in C. rufa, Lygodium scandens and Adiantum caudatum, respectively (Figure 1 b).
Earlier findings showed remarkable increase in the Flavonoids contents of the stressed soybean cultivars. Since
flavonoids inhibit the enzymes responsible for superoxide anion production thus the increase in their values may be
attributed to the protection from free radical induced damage. Similarly proline contents were found to be
accumulated at all the doses of maleic hydrazide in all the fern cultivars- C. rufa, Lygodium scandens and Adiantum
caudatum. A high accumulation (10-60%) of proline contents were observed at the highest dose of mutagen (125
ppm). The accumulation and protective effect of proline has been observed in many higher plants and bacteria as
well as protozoa, algae, and marine invertebrates [5].
Mutagen induced lipid peroxidation of the cellular components in plants were studied by estimating the level of
MDA in treated and untreated plantlets and the related data are depicted in the figure 2 a. The lipid peroxidation in
non- stressed C. Rufa was observed as 1.4609 nmol MDA (mg fresh mass)-1, whereas it was found to be 1.351 and
1.6125 nmol MDA (mg fresh mass)-1. Treated plantlets showed 10-45% increase in total Malondialdehyde contents
as compared to the untreated plants showing high level of lipid peroxidation in maleic hydrazide treated plants.
Similarly the increase in peroxide radical contents was observed to be linearly related with the level of lipid
peroxidation (Figure 2 b). MDA is an intermediate compound produced due to lipid peroxidation, the measurements
of its contents can be used as an index for the injury caused by free radicals produced during oxidative stress. The
results obtained here are in agreement with other authors [1, 10].
CONCLUSION According to the results obtained, it may be concluded that the mutagen-maleic hydrazide affected the overall
growth of all fern species, severely. Decrease in total protein contents and high increase in the level of lipid
peroxidation proved the oxidative damage caused by free radicals formed in response to the mutagen. Increase in
non-enzymatic antioxidants may be attributed to the elevation of natural antioxidant defense system. Initial increase
in the enzymatic activities might be due to the increased activities of stress relief genes and their gene products.
Ferns are good Phytoremediator and can be used to remove heavy metals and other pollutants from the polluted soil.
The study might provide suitable keys for studying the interrelationships of the chemical treatments and plant
defense systems.
Fig.-1: Effect of Maleic hydrazide on protein (a) and non-enzymatic antioxidants (b) in ferns.
(a) (b)
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Vol.1, No.2, 52-55 (2011)
Alok Kr. Singh et al.
55
Fig.-2: Effect of Maleic hydrazide on lipid peroxidation (a) and hydrogen peroxide radicals (b) in ferns.
REFERENCES 1. Agarwal S., Pandey V., Indian Journal of Plant Physiology, 8(3) (2003) 264.
2. Antony R.S., Khan A.E., Thomas, J., Journal of Economic & Taxonomic Botany, 24 (2000) 413.
3. Bates L.S., Waldren R.P., Teare I.D., Plant and Soil, 39(1) (1973) 205.
4. Carrier H.B., Day B.E., Crafts A.S., Bot. Gaz., 112 (1950) 272.
5. Cechin I., Rossi S.C., Oliveira V.C., Fumis T.F., Photosyn., 44(1) (2006) 143.
6. Christopher D.N. et al., International Journal of Environmental Research and Public Health, 7 (2010) 3298.
7. Dai Q. et al., Physiol Plant, 101 (1997) 301.
8. Darlington and McLeish J., Nature, (1951) 167.
9. Halliwell B., and Gutteridge J.M.C., Lancet, 1 (1994) 1396.
10. Hasanuzzaman M. et al., American Journal of Plant Physiology, 5 (2010) 295.
11. Heath R.L., Packer L., Arch Biochem Biophysics, 125 (1968) 189.
12. Hevly, R.H. et al., J Ariz. Acad. Sci., 2 (1963) 164.
13. Hitchcock C.L., Cronquist A., Ownbey M., Thompson J.W., University of Washington Press, Seattle, WA.
(1969) Pp. 914.
14. Lowry O.H., Rosenbrough N.J., Farr A.L., Randall R.J., J. Biol. Chem., 193 (1951) 265.
15. Mirecki R.M., Teramura A.H., Plant Physiol., 74 (1984) 475.
16. Naylor A.W., Davis E.A., Bot. Gaz. ,112 (1950) 112.
17. Rozema J., Boelen P., Blokker P., Environ. Pollut., 137(3) (2005) 428.
18. Sagisaka, S., Plant Physiol ,57 (1976) 308.
19. Schoene D.L., Hoffman O.L., Science, 109 (1949) 588.
20. Sheffield E., Douglas, G.E., Hearned S.J., Huxham S., Wynn, J.M., Amer. Fern J., 91 (2001) 179.
21. Shiu, C.T., and Lee T.M., J. Exp. Bot., 56(421) (2005) 2851.
22. White D.G., Maleic hydrozide. Discovery, II. (1950) 379.
[IJCEPR-158/2011]
(a) (b)
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 56-60
January-April, 2011
V. Bhaskara Raju and A. Lakshmana Rao
Reversed Phase HPLC Analysis of Valsartan in Pharmaceutical Dosage
Forms
V. Bhaskara Raju1 and A. Lakshmana Rao
2*
1Sri Vasavi Institute of Pharmaceutical Sciences, Tadepalligudem- 534 101, A.P., India. 2V.V. Institute of Pharmaceutical Sciences, Gudlavalleru- 521 356, A.P., India. * E-mail: [email protected]
Article History:
Received:8 February 2011
Accepted:27 February 2011
ABSTRACT
A rapid, precise, accurate, specific and sensitive reverse phase liquid chromatographic method has been developed for the
estimation of valsartan in pure and tablet formulation. The chromatographic method was standardized using a Xterra C18 column
(100×4.6 mm I.D., 5 µm particle size) with UV detection at 210 nm and flow rate of 1 ml/min. The mobile phase consisting of a
mixture of phosphate buffer pH 3 and acetonitrile in the ratio of 50:50 v/v was selected. The proposed method was validated for
its sensitivity, linearity, accuracy and precision. The retention time for valsartan was 4.450 min. The % recovery was within the
range between 98.6 % and 101.2 %. The percentage RSD for precision and accuracy of the method was found to be less than 2
%. This method can be employed for routine quality control analysis of valsartan in tablet dosage forms.
Keywords: Valsartan, Estimation, Validation, Tablets, RP-HPLC. ©2011 ijCEPr. All rights reserved
INTRODUCTION Valsartan [1] is a nonpeptide, orally active and specific angiotensin II receptor blocker acting on the AT1 receptor
subtype. Valsartan is chemically described as N-(1-oxopentyl)-N-[[2'-(1H-tetrazol-5-yl)[1,1'-biphenyl]-4-
yl]methyl]-L-valine (Fig. 1) [2]. Angiotensin II is formed from angiotensin I in a reaction catalyzed by angiotensin-
converting enzyme (ACE II). Angiotensin II is the principal pressor agent of the renin-angiotensin system, with
effects that include vasoconstriction, stimulation of synthesis and release of aldosterone, cardiac stimulation and
renal reabsorption of sodium. Valsartan blocks the vasoconstrictor and aldosterone-secreting effects of angiotensin
II by selectively blocking the binding of angiotensin II to the AT1 receptor in many tissues, such as vascular smooth
muscle and the adrenal gland. Its action is therefore independent of the pathways for angiotensin II synthesis. A few
spectrophotometric [3-7], HPLC [8-14], UPLC [15] and LC-MS [16-19] methods
were reported earlier for the
determination of valsartan in bulk and pharmaceutical dosage forms. In the present study the authors report a rapid,
sensitive, accurate and precise HPLC method for the estimation of valsartan in bulk samples and in tablet dosage
forms.
Fig.-1: Chemical structure of valsartan
MATERIALS AND METHOD Chromatographic conditions
The analysis of the drug was carried out on a Waters HPLC system equipped with a reverse phase Xterra C18
column (100x4.6 mm., 5 µm), a 2695 binary pump, a 20 µl injection loop, a 2487 dual absorbance detector and
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Vol.1, No.2, 56-60 (2011)
V. Bhaskara Raju and A. Lakshmana Rao
57
running on Waters Empower software. The UV spectrum of valsartan was taken using a Elico SL-159 UV-Visible
spectrophotometer.
Chemicals and solvents The reference sample of valsartan was supplied by Lupin Pharmaceutical Industries Ltd., Ahmadabad. HPLC grade
water and acetonitrile were purchased from E. Merck (India) Ltd., Mumbai. Potassium dihydrogen phosphate and
orthophosphoric acid of AR grade were obtained from S.D. Fine Chemicals Ltd., Mumbai.
Preparation of pH 3.0 phosphate buffer
Seven grams of KH2PO4 was weighed into a 1000 ml beaker, dissolved and diluted to 1000 ml with HPLC water. 2
ml of triethyl amine was added and pH adjusted to 3.0 with orthophosporic acid.
Preparation of mobile phase and diluents
500 ml of the phosphate buffer was mixed with 500 ml of acetonitrile. The solution was degassed in an ultrasonic
water bath for 5 minutes and filtered through 0.45 µ filter under vacuum.
Procedure
A mixture of phosphate buffer and acetonitrile in the ratio of 50:50 v/v was found to be the most suitable mobile
phase for ideal separation of valsartan. The solvent mixture was filtered through a 0.45 µ membrane filter and
sonicated before use. It was pumped through the column at a flow rate of 1.0 ml/min. The column was maintained at
ambient temperature. The pump pressure was set at 800 psi. The column was equilibrated by pumping the mobile
phase through the column for atleast 30 min prior to the injection of the drug solution. The detection of the drug was
monitored at 210 nm. The run time was set at 7 min. Under these optimized chromatographic conditions the
retention time obtained for the drug was 4.450 min. A typical chromatogram showing the separation of the drug is
given in Fig. 2.
Fig.-2: Typical chromatogram of valsartan
Calibration plot
About 10 mg of valsartan was weighed accurately, transferred into a 100 ml volumetric flask and dissolved in 25 ml
of a 50:50 v/v mixture of phosphate buffer and acetonitrile. The solution was sonicated for 15 min and the volume
made up to the mark with a further quantity of the diluent to get a 100 µg/ml solution. From this, a working standard
solution of the drug (10µg/ml) was prepared by diluting 1 ml of the above solution to 10 ml in a volumetric flask.
Further dilutions ranging from 5-25 µg/ml were prepared from the solution in 10 ml volumetric flasks using the
above diluent. 20 µl of each dilution was injected six times into the column at a flow rate of 1.0 ml/min and the
corresponding chromatograms were obtained. From these chromatograms, the average area under the peak of each
dilution was computed. The calibration graph constructed by plotting concentration of the drug against peak area
was found to be linear in the concentration range of 5-25 µg/ml of the drug. The relevant data are furnished in
Table-1. The regression equation of this curve was computed. This regression equation was later used to estimate the
amount of valsartan in tablet dosage forms.
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Vol.1, No.2, 56-60 (2011)
V. Bhaskara Raju and A. Lakshmana Rao
58
Table-1: Calibration data of the method
Concentration (µg/ml) Mean peak area (n=5)
5 605930
10 1244187
15 1865633
20 2468676
25 3062337
Validation of the proposed method
The objective of the method validation is to demonstrate that the method is suitable for its intended purpose as it is
stated in ICH guidelines [20]. The method was validated for linearity, precision, accuracy, specificity, stability and
system suitability. Standard plots were constructed with five concentrations in the range of 5-25 µg/ml prepared in
triplicates to test linearity. The peak area of valsartan was plotted against the concentration to obtain the calibration
graph. The linearity was evaluated by linear regression analysis that was calculated by the least square regression
method. The precision of the assay was studied with respect to both repeatability and intermediate precision.
Repeatability was calculated from five replicate injections of freshly prepared valsartan test solution in the same
equipment at a concentration value of 100 % (10 µg/ml) of the intended test concentration value on the same
day. The experiment was repeated by assaying freshly prepared solution at the same concentration additionally on
two consecutive days to determine intermediate precision. Peak area of valsartan was determined and precision was
reported as % RSD and the results are furnished in Table-2.
Table-2: Precision of the proposed HPLC method
Peak area Concentration of valsartan (10 µg/ml)
Intra-day Inter-day
Injection-1 1237412 1241721
Injection-2 1238580 1241059
Injection-3 1239480 1242984
Injection-4 1241807 1244489
Injection-5 1244696 1247070
Average 1240395 1243465
Standard Deviation 2895.0 2403.4
% RSD 0.23 0.19
The accuracy of the HPLC method was assessed by analyzing solutions of valsartan at 50, 100 and 150 %
concentrated levels by the proposed method. The results are furnished in Table-3. The system suitability parameters
are given in Table-4.
Table-3: Accuracy studies
Concentration Amount added (mg) Amount found (mg) % Recovery % Mean recovery
50 % 5.02 4.95 98.6 %
100 % 10.1 10.17 100.7 %
150 % 15.1 15.28 101.2 %
100.2 %
Table-4: System suitability parameters
Parameter Result
Linearity (µg/ml) 5-25
Correlation coefficient 0.9998
Theoretical plates (N) 4547
Tailing factor 1.20
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Vol.1, No.2, 56-60 (2011)
V. Bhaskara Raju and A. Lakshmana Rao
59
LOD (µg/ml) 0.012
LOQ (µg/ml) 0.040
Estimation of valsartan in tablet dosage forms
Two commercial brands of tablets were chosen for testing the suitability of the proposed method to estimate
valsartan in tablet formulations. Twenty tablets were weighed and powdered. An accurately weighed portion of this
powder equivalent to 25 mg of valsartan was transferred into a 100 ml volumetric flask and dissolved in 25 ml of a
50:50 v/v mixture of phosphate buffer and acetonitrile. The contents of the flask were sonicated for 15 min and a
further 25 ml of the diluent was added, the flask was shaken continuously for 15 min to ensure complete solubility
of the drug. The volume was made up with the diluent and the solution was filtered through a 0.45 µ membrane
filter. This solution was injected into the column six times. The average peak area of the drug was computed from
the chromatograms and the amount of the drug present in the tablet dosage form was calculated by using the
regression equation obtained for the pure drug. The relevant results are furnished in Table-5.
Table-5: Assay and recovery studies
Formulation Label claim (mg) Amount found (mg) % Amount found
Formulation 1 40 40.12 99.70
Formulation 2 40 39.86 100.35
RESULTS AND DISCUSSION Selection of the detection wavelength The UV spectra of valsartan in 50:50 v/v mixture of phosphate buffer and acetonitrile was scanned in the region
between 200 and 400 nm and shows λmax at 210 nm.
Optimization of the chromatographic conditions
Proper selection of the stationary phase depends upon the nature of the sample, molecular weight and solubility.
Mixture of phosphate buffer and acetonitrile was selected as mobile phase and the effect of composition of mobile
phase on the retention time of valsartan was thoroughly investigated. The concentration of phosphate buffer and
acetonitrile were optimized to give symmetric peak with short run time. A short run time and the stability of peak
asymmetry were observed in the ratio of 50:50 % v/v of phosphate buffer and acetonitrile. It was found to be
optimum mobile phase concentration. In the proposed method, the retention time of valsartan was found to be 4.45
min. Quantification was linear in the concentration range of 5-25 µg/ml. The regression equation of the linearity plot
of concentration of valsartan over its peak area was found to be Y=8161.7+122746.06X (r2=0.9998), where X is the
concentration of valsartan (µg/ml) and Y is the corresponding peak area. The number of theoretical plates calculated
was 4547, which indicates efficient performance of the column. The limit of detection and limit of quantification
were found to be 0.012 µg/ml and 0.040 µg/ml respectively, which indicate the sensitivity of the method. The use of
phosphate buffer and acetonitrile in the ratio of 50:50 v/v resulted in peak with good shape and resolution. The high
percentage of recovery indicates that the proposed method is highly accurate. No interfering peaks were found in the
chromatogram of the formulation within the run time indicating that excipients used in tablet formulations did not
interfere with the estimation of the drug by the proposed HPLC method.
CONCLUSION The proposed HPLC method is rapid, sensitive, precise and accurate for the determination of valsartan and can be
reliably adopted for routine quality control analysis of valsartan in its tablet dosage form.
ACKNOWLEDGEMENTS The authors are thankful to M/s Lupin Pharmaceutical Industries Ltd., Ahmadabad, for providing a reference sample
of valsartan.
REFERENCES 1. Budavari S. The Merck index, Merck and Co. Press: Whitehouse Station, NJ, 12th Edn, 1997.
2. www.rxlist.com
3. Gupta K.R., Wadodkar A.R., Wadodkar S.G., International Journal of Chem Tech Research, 2 (2010) 985.
4. Gupta K.R., Mahapatra A.D., Wadodkar A.R., Wadodkar S.G., International Journal of Chem Tech Research, 2
(2010) 551.
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Vol.1, No.2, 56-60 (2011)
V. Bhaskara Raju and A. Lakshmana Rao
60
5. Tatar S., Saglik S., Journal of Pharmaceutical and Biomedical Analysis, 30 (2002) 371.
6. Nevin E., Analytical Letters, 35 (2002) 283.
7. Satana E., Altınay S., Goger N.G., Sibel A., Ozkan S.A., Senturk Z., Journal of Pharmaceutical and Biomedical
Analysis, 25 (2001) 1009.
8. Kul D., Dogan-Topal B., Kutucu T., Uslu B., Ozkan S.A., Journal of AOAC International, 93 (2010) 882.
9. Patro S.K., Kanungo S.K., Patro V.J., Choudhury N.S.K., E-Journal of Chemistry, 7 (2010) 246.
10. Brunetto M.R., Contreras Y., Clavijo S., Torres D., Delgado Y., Ovalles F., Ayala C., Gallignani M., Estela
J.M., Martin V.C., Journal of Pharmaceutical and Biomedical Analysis, 50 (2009) 194.
11. Chitlange S.S., Bagri1 K., Sakarkar D.M., Asian Journal of Research in Chemistry, 1 (2008) 15.
12. Kocyigit-Kaymakcoglu B., Unsalan S., Rollas S., Pharmazie, 61 (2006) 586.
13. Macek J., Klima J., Ptacek P., Journal of Chromatography, B 832 (2006) 169.
14. Daneshtalab N., Lewanczuk R.Z., Jamali F., Journal of Chromatography, B 766 (2002) 345.
15. Krishnaiah CH., Raghupathi Reddy A., Ramesh Kumar., Mukkanti K., Journal of Pharmaceutical and
Biomedical Analysis, 53 (2010) 483.
16. Sampath A., Raghupathi Reddy A., Yakambaram B., Thirupathi A., Prabhakar M., Pratap Reddy P., Prabhakar
Reddy V., Journal of Pharmaceutical and Biomedical Analysis, 50 (2009) 405.
17. Selvan P.S., Gowda K.V., Mandal U., Solomon W.D.S., Pal T.K., Journal of Chromatography, B 858 (2007)
143.
18. Li H., Wang Y., Jiang Y., Tang Y., Wang J., Zhao L., Gu J., Journal of Chromatography, B 852 (2007) 436.
19. Koseki N., Kawashita H., Hara H., Niina M., Tanaka M., Kawai R., Nagae Y., Masuda N., Journal of
Pharmaceutical and Biomedical Analysis, 43 (2007) 1769.
20. ICH, Q2B. Validation of analytical procedures methodology, In Proceedings of The International Conference
on Harmonization, Geneva 1993.
[IJCEPR-150/2011]
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International Journal of Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and Chemical, Environmental and
Pharmaceutical ResearchPharmaceutical ResearchPharmaceutical ResearchPharmaceutical Research
Vol. 2, No.1, 61-66
January-April, 2011
Haritha Meruvu , Meena Vangalapati
Nattokinase :A Review on Fibrinolytic Enzyme
Haritha Meruvu , Meena Vangalapati* Center for Biotechnology, Department of Chemical Engineering, College of Engineering,
Andhra University, Visakhapatnam–530 003, Andhra Pradesh, INDIA.
Email: [email protected] Article History:
Received:26 April 2011
Accepted:28 April 2011
ABSTRACT
Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases. It is extracted and
highly purified from a traditional Japanese food called Natto which is made from fermented soybeans, Glycine max (L.)
Merr.Natto is produced by a fermentation process by adding Bacillus subtilis, subsp. Natto to boiled soybeans. The ensuing
Nattokinase enzyme is produced when Bacillus subtilis acts on the soybeans. Nattokinase has caused natto wide attention around
the world Natto is not only unique flavor and rich nutrition, but also due to a variety of health functions, known as "super health
food”. The present review attempts to encompass the up-to-date comprehensive literature analysis on Nattokinase with respect to
its properties, source and its various medical uses.
Keywords: Nattokinase, Natto, Glycine max (L.), Bacillus subtilis subsp. Natto. ©2011 ijCEPr. All rights reserved
INTRODUCTION Nattokinase was discovered in 1980 by Dr Hiroyuki Sumi, researcher at Chicago University after testing over 173
natural foods as potential thrombolytic agents, searching for a natural agent that could effectively dissolve thrombus
allied with cardiac and cerebral infarction [3-5]. Nattokinase was discovered in Natto, a fermented cheese-like food
that has been used in Japan for over 1000 year. Natto is a traditional Japanese food made of soybeans . To prepare
the beans are cooked and then by the action of the bacterium Bacillus subtilis ssp.natto fermented [8,27] .During this
process is formed a slimy, stringy substance to the beans. In the traditional method of preparation are the bacteria
from rice straw , into which the beans are wrapped [3] . In the modern manufacturing process, the bacteria cultures
inoculated with beans, so that the use of rice straw is no longer necessary [21]. The botanical source for
Nattokinase is Glycine max(L. )Merr. It appears as a yellow-white fine powder [22].
Fig.-1: Natto, traditionally wrapped in rice straw
Natto is considered a very healthy food; a health product in the fermentation is some evidence for emerging
substances [46]. Nattokinase is used for cardiovascular diseases including heart disease, high blood
pressure, stroke, chest pain (angina), deep vein thrombosis,, “hardening of the arteries”
(atherosclerosis), hemorrhoids, varicose veins, poor circulation,and peripheral artery disease[43-45]. It is also used
for pain, fibromyalgia, chronic fatigue syndrome, endometriosis, uterine fibroids, muscle spasms, infertility,
cancer, and a vitamin-deficiency disease called beriberi [40].
Properties
Nattokinase is a fibrinolytic enzyme, meaning that it breaks down fibrin, an insoluble white protein produced by the
conversion of fibrinogen (a protein in the plasma of blood for clotting) by thrombin (a blood clotting enzyme).
Nattokinase is a serine protease with 275 amino acid residues and a molecular weight of 27,728 Daltons.
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Vol.1, No.2, 61-66 (2011)
Haritha Meruvu , Meena Vangalapati
62
Nattokinase has a high homology with the subtilisin enzymes and DNA sequencing shows 99.5 and 99.3%
homology to subtilisin E and amylosacchariticus respectively [36, 38-39]. Nattokinase degrades fibrin clots both
directly and indirectly. Nattokinase degrades fibrin directly in clot lysis assays with activity comparable toplasmin.
Kinetic assays suggest that it is 6 times more active than plasmin in degrading cross-linked fibrin. Nattokinase
degrades fibrin indirectly by affecting plasminogen activator activity. The other names of nattokinase are BSP, Natto
Extract, Nattokinasa, NK, Fermented Soybeans, Soy Natto and Subtilisin NAT. Below, is the chemical structure of
nattokinase [26-28].
Fig.-2: Chemical structure of nattokinase
Characteristics of nattokinase [15, 17, 19-21]
Protien(Enyme) names Recommended name: Subtilisin NAT, EC=3.4.21.62
Alternative name(s):Nattokinase,cardiokinase,
Gene names Name: aprN
Organism Bacillus subtilis subsp. natto
Taxonomic identifier 86029 [NCBI]
Taxonomic lineage Bacteria › Firmicutes › Bacillales › Bacillaceae › Bacillus
Protein attributes
Sequence length 381 AA.
Sequence status Complete.
Sequence processing The displayed sequence is further processed into a mature form.
Protein existence Evidence at protein level.
General Annotation
Function Subtilisin is an extracellular alkaline serine protease, it catalyzes the hydrolysis of
proteins and peptide amides. Subtilisin NAT also has fibrinolytic activity.
Catalytic activity Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for
a large uncharged residue in P1. Hydrolyzes peptide amides.
Subunit structure Monomer.
Subcellular location Secreted.
Sequence similarities Belongs to the peptidase S8 family.
Biophysicochemical
properties
Kinetic parameters:
KM=0.48 mM for Suc-Ala-Ala-Pro-Phe-pNA
Researches suggest that Nattokinase may promote normal blood pressure, reduce whole blood viscosity and increase
circulation being an effective supplement to support cardiovascular health[19].Its strong thrombolytic activity
promotes arterial health both directly, dissolving existing thrombus, and indirectly, enhancing body’s production of
plasmin and urokinase by a direct cleavage of plasminogen activator inhibitor [32-35]].
The human body produces several types of enzymes for making thrombus, but only one main enzyme for breaking it
down and dissolving it - plasmin. Nattokinase has plasmin-like bio-characteristic that lyses fibrin directly or
indirectly in three different pathways [9, 10, 27]:
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Vol.1, No.2, 61-66 (2011)
Haritha Meruvu , Meena Vangalapati
63
1. Nattokinase lyses fibrin directly.
2. Nattokinase enhances plasmin through active pro-urokinase (endogenous).
3. t-PA (Tissue Plasminogen Activators) is like urokinase and active plasmin,Nattokinase increases the
concentration of t-PA.
Fig.-3: The physiological effects of nattokinase on fibrin
Source
The botanical source for Nattokinase extraction is Glycine max(L. )Merr. It is a dicotyledonous annual herb
belonging to fabaceae with common names wild soybean and reseeding soybean; with synonyms Dolichos soja L.,
Glycine gracilis Skvortzov, Glycine hispida (Moench) Maxim., Glycine soja (L.) Merr., nom. illeg., non Glycine soja
Siebold & Zucc., Glycine ussuriensis Regel & Maack, Phaseolus max L., Soja hispida Moench, Soja max (L.) Piper
[22-26 ].
Fig.-4: Images for Glycine max (L.) Merr
Classification: Glycine max (L.) Merr.
Kingdom Plantae – Plants
Subkingdom Tracheobionta – Vascular plants
Superdivision Spermatophyta – Seed plants
Division Magnoliophyta – Flowering plants
Class Magnoliopsida – Dicotyledons
Subclass Rosidae
Order Fabales
Family Fabaceae – Pea family
Genus Glycine Wild. – soybean
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Species Glycine max (L.) Merr. – soybean
Table-1: Compositional analysis of soybean [4]
Raw soybean Boiled soybean
pH 6.4 6.9
Moisture 10.49 68.42
Ash 4.22 1.06
Crude fibre 14.22 0.08
Protien 32.92 12.15
Fat 8.19 1.01
Reducing sugar(g/l) 0.29 0.22
Ammonia(g/100ml) 0 0
The microbial source for nattokinase extraction is Bacillus subtilis subsp. Natto. The other name is Bacillus subtilis
var. natto. This bacterium is used to produce natto by fermentation. Cooked soy beans are inoculated with the
bacterial starter culture. The fermentation process takes at room temperature in a day; this time may be reduced to
eight hours to six, if the temperature is increased from 40° C to 43 ° C. The maximum temperature reached during
the fermentation process should be is 50 ° C; above which the fermentation stops as the bacteria die [11, 16, 17].
Fig.-5: Natto: Fermented soybean
Bacterial Classification
› Firmicutes
› Bacilli
› Bacillales
› Bacillaceae
› Bacillus
› Bacillus subtilis group
› Bacillus subtilis
Fig.-6: Bacillus subtilis Natto observed under microscope
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The various strains of Bacillus subtilis subsp. Natto are
1. CCRC 14716
2. IAM 1028
3. IAM 1163
4. IAM 1232
5. NC2-1
6. NR-1
7. OK2
Microbial fermentation is carried out using substrates like soybean, wheat bran, shrimp shell. These are the three
substrates that are efficient in production of nattokinase enzyme upon fermentation. The essential components for
fermentation are listed in comparison with three substrates [6-11].
Table-2: Compositional analysis of various substrates
Constituents
(%)
Soybean meal Wheat bran Shrimp shell meal
Fibre 7 30 19
Moisture 8 14 14
Protein 43 16 35
nitrogen 7 14 8
lipid 7 4 9
carbohydrate 27 6 3
Medical uses In the main nattokinase works to support healthy blood circulation in two different ways. First off, nattokinase
resembles plasmin, so it can break down fibrin directly. Secondly, nattokinase enhances the body’s natural
production of plasmin, which also helps to break down fibrin. [2,11] In a nutshell, Nattokinase:
• Supports normal circulation, blood flow, and blood viscosity (thickness)
• Supports the body’s normal blood-clotting mechanism
• Supports the body’s production of plasmin, which reduces fibrin
• Helps to maintain normal blood pressure levels
Nattokinase is used for cardiovascular diseases including heart disease, high blood pressure, stroke, chest
pain (angina), deep vein thrombosis, hardening of the arteries (atherosclerosis), hemorrhoids, varicose veins, poor
circulation, and peripheral artery disease stroke, venous stasis, thrombosis, emboli, fibromyalgia/chronic
fatigue, claudication, retinal pathology, hemorrhoid, varicose veins, soft tissue rheumatisms, muscle spasm, poor
healing[31-35]. .
It is also used for pain, fibromyalgia, chronic fatigue syndrome, endometriosis, uterine fibroids, muscle
spasms, tissue oxygen deprivation, infertility and cancer. Because nattokinase is an edible enzyme and is been used
as nutrient supplement, it can be used to digest amyloids in body. Nattokinase can be used to remove infectious
prion from animal feed, surgical instrument, and blood product [3-4, 44].
CONCLUSION Nattokinase is a potent fibrinolytic enzyme discovered in the extract of natto and produced via fermentation
of Bacillus subtilis natto from boiled soybean.The safety record of its potent fibrinolytic enzyme, Nattokinase, is
based upon the long term traditional use of the food, and recent scientific studies. Nattokinase has many benefits
including its prolonged effects, cost effectiveness, and its ability to be used preventatively. It is a naturally
occurring, food-based dietary supplement that has demonstrated stability in the gastrointestinal tract, as well as to
changes in pH and temperature. Stressful era of modernization has led to high rates of cardiovascular diseases;
thence it would then seem prudent to add this effective natural product to our heart health preventive arsenal as more
recently, both clinical and non-clinical studies have demonstrated that Nattokinase supports heart health and
promotes healthy circulation. Hereby, this paper paraphrases the properties, biological activity and the botanical and
microbial sources of nattokinase. Moreover, the assorted therapeutic and medicinal uses are also summed in
herewith.
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REFERENCES 1. Cesarone M.R., Belcaro G., Nicolaides A.N., Angiology, 54 (2003) 531.
2. Chen P.T., Chao Y.P., Biotechnol. Lett., 28 (2006) 1595.
3. Deepak V., Bioresource Technol. ,99 (2008) 8170.
4. Deepak V.,Kalishwaralal, Bioresour. Technol., 99 (2008) 8170.
5. Dobrovolsky A.B.,Titaeva E.V.,Biochemistry Biokhimiia, 67 (2002) 99.
6. Fujita M., Hong K., Ito Y., Fujii R., Biol Pharm Bull, 18 (1995) 1387.
7. Fujita M., Nomura K., Nishimuro S., Biochem Biophys Res Commun, 197 (1993) 1340.
8. Fujita M., Biochemical and biophysical Research Communications, 197 (1993) 1340.
9. Hsia C.H., Shen M..C, Lin J.S., Nutr Res , 29 (2009) 190.
10. Hsia C.H., Nutr. Res., 28 (2008) 161.
11. Hsu R.L., Lee K.T., Wang J.H., Lee L.Y., J Agric Food Chem, 57 (2009) 503.
12. H w a n g K . J . , C h o K . H . , J o u r n a l o f M i c r o b i o l o g y a n d B i o t e c h n o l o g y , 1 7 ( 2 0 0 7 ) 1 4 6 9 .
13. Kim, Appl. Environ. Microbiol., 62 (1996) 2482.
14. Kim S., Choi N.S., Biosci. Biotechnol. Biochem., 64 (2000) 1722.
15. Kim J.Y., Gum S.N., Hypertens Res., 31 (2008) 1583.
16. Kim S. B.,Lee, D., J. Ind. Microbiol. Biotechnol., 33 (2006) 436.
17. Kim W., Choi K., Appl. Environ. Microbiol. 62 (1996) 2482.
18. Kim H.K., J. Biosci. Bioeng., 4 (1997) 307.
19. Kim W., Choi K., Applied and Environmental Microbiology, 62 (1996) 2482.
20. Ko, J., Yan J., Process Biochem., 44 (2009) 70.
21. Ku T.W., J. Agric. Food Chem., 55 (2008) 271.
22. Ku TW., Tsai RL., Pan TM,. J Agric Food Chem, 57 (2009) 292.
23. Liu X.L., Du L.X., Microbiol. Biotechnol., 67 (2005) 209
24. Liu J., Xing, J., Chang T., Process Biochem., 40 (2005) 2757.
25. MineY.,Wong K,Food Research International, 38 (2005)243
26. Omur a KHito sug i M, Zhu XI keda MJo ur na l o f Pha r maco lo gica l Sc iences , 99 ( 2005) 247 .
27. P a i s E . , A l e x y T . , C l i n H e m o r h e o l M i c r o c i r c , 3 5 ( 2 0 0 6 ) 1 3 9 .
28. P eng Y . , Yang X . , Zhang Y . , Ap p l ied M icro b io lo gy and B io techno lo gy, 6 9 ( 200 5) 1 26 .
29. Peng Y., Huang Q., Biochem. Physiol, 134 (2003) 45.
30. Shieh C.J., Phan Thi, L.A., Shih I.L., Biochem. Eng., J 43 (2009) 85.
31. Sugimoto S., Fujii T., Morimiya T., Biosci Biotechnol Biochem, 71 (2007) 2184.
32. Sumi H., Hamada H., Nakanishi K., Acta Haematol., 84 (1990) 139.
33. Sumi H., Hamada H., Tsushima H., Mihara H., Muraki H., Experientia, 43 (1987) 1110.
34. S u m i H . , N a k a j i ma N . , Y a t a g a i C . , B i o c h e m i s t r y & M o l e c u l a r B i o l o g y , 1 ( 1 9 9 5 ) 5 4 3 .
35. Sumi H.,Hamada H., Experientia, 43(1987)1110.
36. Suzuki Y, Kondo K, Ichise H, Nutrition, 19 (2003) 261.
37. Suzuki Y., Kondo K., Matsumoto Y., Urano T., Life Sci., 73 (2003) 1289.
38. Tai MW., Sweet BV., Am J Health Syst Pharm, 63 (2006) 1121.
39. Wang C., Du M., Zheng D., J Agric Food Chem, 10 (2009) 1021.
40. Wang C. T.,Ji B.P., World J. Microbiol. Biotechnol., 22 (2006) 1365.
41. W a n g C T , J o u r n a l o f I n d u s t r i a l M i c r o b i o l o g y B i o t e c h n o l o g y , 3 3 ( 2 0 0 6 ) 7 5 0 .
42. Wang D.S., J. Food Process Eng., 29 (2006) 22.
43. Weng M., Zheng Z., Bao W., Cai Y., Biochim Biophys Acta, 1794 (2009) 1566.
44. Wu D.J., Acta Cardiol. Sinica, 25 (2009) 26.
45. Yang N.C., Chou C.W., Chen C.Y., Asia Pac J Clin Nutr, 18 (2009) 310.
46. Yoshinori M, Food Res. Int. ,38 (2005) 243 .
[IJCEPR-152/2011]
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