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Online International Interdisciplinary Research Journal, {Bi-Monthly}, Volume-II, Issue-II, Mar-Apr2012
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“Ultrasonic Wave technique utilize for study the behavior of peptide-
Carnosine in a different solvents”
Dr. R.B. Pawar Department of Chemistry, S.S.S K.R. Innani Mahavidyalaya Karanja (Lad), Dist. Washim (M.S.),
India.
ABSTRACT
The ultrasonic velocity of carnosine in different solvent mixtures has been
investigated to understand the effect of carnosine in interaction with water and organic
solvent. The various parameters such as molality, D0, d0, U0, US, DS, d0, BS, B0, K(S), φv
determined as well as the results have been discussed.
KEYWORDS: Apparent molal adiabatic compressibility, Hydration numbers.
Introduction
Adiabatic compressibility, Viscosity, Refractive index, apparent molal volume etc
are physical properties of liquid. Vibrational waves of frequency above hearing range of
normal ear are referred as ultrasonic waves. All waves of frequencies more than 20 KHz
are ultrasonic waves.
Ultrasonic waves, in recent years, have acquired the status of an important probe
for the study of structure and properties of matter in basic science. In the field of
technology, the waves are being used for detection of flaws, testing of materials,
mechanical cleaning of surface etc. In medical science too, the waves are being used to
detect bone fractures, cancer tumors, foetal conditions and in physiotherapy, bloodless
surgery, cardiology, gynacology etc. Present day applications of ultrasonic are emerging
in the field of forensic science, space research and in wars. Adiabatic compressibility and
apparent molal compressibility have been used to study the relative association, specific
constant factor and solvation number of the system.
The study of molecular interactions in liquids provides valuable information
regarding internal structure, molecular association, complex formation, internal pressure
etc. The various techniques available to study them are Nuclear Magnetic Resonance,
Microwave, Ultraviolet & Scattering and Ultrasonic investigation. Nuclear magnetic
resonance technique reflects the effect on the proton bearing molecules, whereas
microwave absorption provides information through dielectric constant. Neutron and X-
ray scatterings help in the study of molecular motion. The spectroscopic techniques
provide useful information of interactions when the interaction energies involved are
large. Weak molecular interactions cannot be resolved from the observed spectra.
Ultrasonic technique reveals very weak intermolecular interactions due to its useful
wavelength range. Moreover, ultrasonic parameters are directly related to a large number
of thermodynamic parameters. Since various molecular theories of liquid state are based
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on thermodynamic considerations, ultrasonic absorption study and ultrasonic velocity
determination provide means to study them.
For a long time, an important application of ultrasonic velocity measurement in
liquid is to evaluate an adiabatic compressibility and ratio of specific heats. In recent
years, determination of ultrasonic velocity and absorption coefficient have furnished
methods for studying molecular and structural properties of liquids since there exists
intimate relationship between ultrasonic velocity and chemical or structural
characteristics of molecules of liquid. Ultrasonic velocity gives properties of basic
importance to sound velocity in molecular theory of liquid.
Upto 1930, data on velocity of sound in very few liquids were available. The
discoveries of interferometry and optical diffraction method improved the investigation
manifold, both qualitatively and quantitatively.
In recent years, ultrasonic velocity and absorption studies in case of electrolyte
solutions, have led to new insight into the process of ion association and complex
formation. (Kor S.K. et al : Ind. J.Pure Appl. Phy., 7, 784(1969). Soitkar V.S. et al :
Acoustic Lett., 7, 191 (1984).
Number of workers such as (Satyvati
A.V.-Acoustica, 70, 40 (1984).,
(Ramchandran K.- Ind. J.Pure. Appl. Phy., 6, 75(1968)., (Prakash S. et al :
Ind.JChem.8,489(1964). (Marks, G.W. : J.Acost.Soc.Am Erica, 38, 327(1960) Agrawal
et al : Acoust. J. Phys., 31, 567 (1978). and Tabhane V.A. :Acoustic Lett., 6, 120
(1983),Tabhane V.A., et al : Ind. J. Pure and Appl. Phys., 23, 502(1985) have made
ultrasonic study of electrolytic solutions and discussed about the variation of ultrasonic
velocity with ion concentration. It has already been established that lowering of
compressibility and an increase in ultrasonic velocity wit reference to that of water, are
proportionate to the number of ions existing in that medium. Most of the ultrasonic work,
in nonaqueous systems, possess an interpretation of solute-solvent interaction (Prasad N.,
et al : Ultrasonics, 18, 160 (1980).
Solvation numbers have been obtained from the study of nonaqueous solutions by
Prakash et al : J. Acoust. Soc., Ind., 4, 39 (1976). Chaturvedi C.V. et al :Ind. J. Chem., 9,
1138 (1970).
Many attempts have been made, in recent years, to study molecular interaction in
pure and binary liquid mixtures Delmus G.T.:Trans. Faraday Soc., 70, 590 (1975). and
various equations of states Barker J.A. et al : J. Mol. Phys., 21, 187 (1971). (D. Gupta A.
et al : Ind. J. Pure and Appl. Phy., 26, 340(1988). for hard sphere fluid have come forward.
Gopalrao R.V. et al :Ind. J. Pure and Appl. Phys., 1 905 (1976). formulated the equation
of state for a square well fluid and obtained some thermodynamic parameters by
extending Flory's equation (Flory P.J., et al :J. Am. Chem. Soc., 86, 3507 (1964) to
mixtures of unrelated types of molecule. (Sharma B.K. : J.Pramana, 14, 477 (1980)
tested the validity of an equation of state of real fluid and determined the expression for
various acoustical parameters to relate them with Gruneisen parameters. (Tabhane V.A.
et al : Ind. J. Pure and Appl. Phys., 33, 248 (1995) have investigated the cluster approach
to thermodynamic behaviour of liquid mixtures of acrolein in methanol, cyclohexane and
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dioxane using Khasare's equation of state (Khasare S.B. : Ind. J. Pure and Appl.Phys.,31,
224 (1993).
The determination of the volume of a solution at different concentrations is very
important because it helps to calculate the properties like apparent molal volume (v) and
apparent molal expansibility (E). Apparent molal volume (v) is a difference between the
volume of solution containing one mole of substance and the volume of contained
solvent.
Ultrasonic parameters are being extensively used to study molecular interactions
in pure liquids (Sheshgiri K. et al :J. Acoustica, 29, 59 (1973). Reddy K.C., liquid
mixtures Sheshagiri K. et al :Ind. J.Pure and Appl.Phys.,9,169 (1971)
and electrolyte solutions (Gnananba S. et al :Ind. J. Pure Appl. Phys., 7, 468 (1969).
Representation in terms of the measured parameters such as velocity of existence of an
interaction, does not provide any interaction about it.
Prigogine I. et al : J. Chem. Phys., 24, 518 (1956). have shown that the excess
parameter such as excess volume (VE) gives interaction on the relative strength of AA,
AB, and BB interactions in the mixture of A and B liquids.
The apparent and partial molal volumes of electrolyte solutions have
proved a very important tool in elucidating the structural interactions i.e. ion-ion, ion-
solvent and solute-solvent interactions occurring in solution.
Recently there has been an increased interest in the state of water in the
living cell. Since most biological macromolecules are physiologically active in aqueous
solutions, knowledge of water-proton interaction is necessary to understand the role or
water solvated to soluble organics in the living cell. A better understanding of this type of
interaction may be obtained from dipolar ions.
Partial molal volume and adiabatic compressibility properties reflect
structural interactions with water molecules or organic solvent molecules and therefore
carnosine a dipeptides is selected for this investigation. For a long time, an important
application of ultrasonic velocity measurement in liquid is to evaluate an adiabatic
compressibility and ratio of specific heats. In recent years, determination of ultrasonic
velocity and absorption coefficient have furnished methods for studying molecular and
structural properties of liquids since there exits intimate relationship between ultrasonic
velocity and chemical or structural characteristics of molecules of liquid. Ultrasonic
velocity gives properties of basic importance to sound velocity in molecular theory of
liquids.
In recent years, ultrasonic velocity and absorption studies in case of
electrolyte solutions, have led to new insight in to the process of ion association and
complex formation (Kor.S. K. et al :- Ind. J. Pure, & Apol Phy.7784 (1969),( Soitkar V.S.
et al 1984). In 1978, millerio and coworkers (Millerio F.J. et.al. 1978) have investigated
the apparent molal volume and adiabatic molal compressibility of 15 amino acids in
aqueous medium. Hydration numbers are calculated using partial molal volume and
adiabatic compressibility data. This particular fields is attracting the attention of several
workers in our country, which can be judged from recent publications (Kaulugud M.V. et
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al 1975) (Singh S. et al 1977) (Reddy K.S. Sreenivasulam and Naidu P.R 1981) (Prakash
O. et al 1982) (Ragouramane D et al 1998) in this field. Some peptides such as Glycil-
Glycine, L-Alanyl-L-alamine DL-Alanyl-DL Phenyl alanine and DL-Alanyl-Glycine
have been studied ( Khobragade B.G, 1999). Adiabatic molal Compressibility and
apparent molal volumes of many electrolytes in mixed organic solvent are found out
earlier. But compressibilities and apparent molal volumes of peptides in aqueous as well
as in water-organic solvent mixtures are not studied so far. Therefore, the present work is
undertaken to make a systematic study of adiabatic molal compressibilities and apparent
molal volume of i) Carnosine (L1) {in-w-225.20} [C9H13N4O3] in ethanol-water.
methanol-water & acetone-water mixtures. Carnosine is a dipeptides of β-alanine &
histidine which are water soluble dipeptides of voluntary muscles.
Experimental
Solvents and chemicals
Instruments
Pyknometer:
Pyknometer (Borosil make) are used in the present investigation for measuring
the densities.
Balance:
'K' Roy one pan electronic balance reading up to5th place of decimals is used for
all weighings. Accuracy of balance was ± 1.00 x 10-5
g.
Ultrasonic Interferometer:
Ultrasonic Interferometer from Mittal Enterprises Model F-80 with accuracy up
to ± 0.03% and frequency 2MHZ. is used for the measurement of ultrasonic velocities of
different solutions.
Thermostat:
A special thermostatic arrangement was done for density and ultrasonic velocity
measurements. Thermostatic water bath (capacity 10-liters) supplied by Yarco Company
having continuous stirring of water was carried out with the help of electric stirrer.
The solvent & chemicals used are prepared & purified by different purification methods
as below.
Acetone
Impurities in acetone are methanol and acetic acid (organic impurities) (less than
0.1 percent) and water (as high as 1 percent).
Purification : One hundred grams of finely powdered sodiumiodides were dissolved
under reflux in 440g of boiling acetone (E. Merck (India) Ltd.) and the solution was
cooled in a mixture of ice and salt (-8°C). The crystals were filtered off and quickly
transferred to a dry distilling cooled in ice. Upon gentle warming, the acetone distilled
rapidly. (Acetone had b.p. 56°C)((a)BS 3978 1966: water for laboratory use. London;
British Standards Institution.(b)D1193-70 (1970). Standard Specification for Reagent
Water. Easto, Md.;American Society for Testing Materials.(c)Reagent Chemicals;
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Supplement 1(1969). Washington, DC; American Chemical Society Publications.)
(Common Apparatus and Basic Techniques 3, 48 Vogel Book).
Ethanol
Rectified is the constant boiling point mixture which ethanol forms with water
and usually contains 95.6 percent of ethanol by weight. (Flory P.J. et al:
J.Am.Chem.Soc., 86, 3507 (1964).
Purification:
1) Ethanol of 99.5 percent purity may be prepared by the dehydration of rectified spirit
with calcium oxide.
A mixture of 250g of calcium oxide (freshly ignited) and 1-litre of rectified spirit
taken into a 2-litre round-bottomed flask fitted with a double surface condenser carrying
a calcium chloride guard-tube was allowed to cool. The mixture was refluxed gently for a
6 hours and allowed to stand overnight. The ethanol was distilled gently discarding the
first 20 ml of distillate into a receiver flask with side arm receiver adapter protected by
means of a calcium chloride guard-tube. The absolute ethanol (99.5%) was preserved in a
glass bottle with a well-fitting stopper.
Methanol
A purity of 99.85 percent with not more than 0.1 percent by weight of water and
not more than 0.02 percent by weight of acetone is claimed in methanol. (Flory P.J. et al
J.Am.Chem.Soc.86, 3507(1964)
Purification:
1) Most of the water was removed from 1-litre of methanol (Methanol Extrapure
(s.d. fine-CHEM Ltd.)) by distillation through fractionating column.
2) Anhydrous methanol was obtained from the fractionally distilled solvent by
treatment with magnesium metal using the procedure given for 'super dry' ethanol.
3) Small proportion of acetone was removed by the following procedure (Morton
and Mark, 1934):
A mixture of 500 ml methanol, 25 ml of furfural and 60 ml of 10 percent sodium
hydroxide solution was refluxed in a 2-litre round-bottomed flask, fitted with a double
surface condenser, for 12 hours. A resin was formed which carried down all the acetone
present. The alcohol was then fractionated, the first 5 ml of which containing a trace of
formaldehyde being rejected. (Methanol had b.p. 65°C)
The sound velocities of peptides i.e. ligands (L1,) are measured in the
concentration range of 0.0072 molality in different percentage of ethanol-water,
methanol-water and acetone-water mixtures. The cell of ultrasonic interferometer was
filled fully with the solution and needle of ammeter was adjusted in the range of 20 to 60
with the help of „adj‟ knob. It was warmed for 10 minutes so that the range should remain
steady. Micrometer reading was noted. Screw was moved anticlockwise to get the
maximum deflections of needle. To movement of screw was co untied to gate 5
deflection. After retuning back to its original position , Micrometer screw was noted.
The difference between these two readings gave the distance travelled by screw for
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getting five maxima. From this distance required though which micrometer screw should
move for one maxima was calculated by dividing it by 5. Same procedure repeated many
times. The observations of systems are represented in table (1) to (2) in table (3) values of
φK(s) & φv which is calculated as follows:-
The apparent molal volume (φv) and apparent molar adiabatic compressibility
(φK(s)) of peptides in solution are determined from density (ds) and adiabatic
compressibility (Bs) of solution using following equations.
φ K(s) = ( )
……… (1)
φ v = ( )
……… (2)
Where M is the molecular weight of solute, m is molality of the solution, do is
the density of solvent, ds is the density of solution, B0 is adiabatic compressibility of
solvent.
The adiabatic compressibility is calculated from ultrasonic velocity using the
Equation: - Bo =
……… (for solvent)
Bs =
……… (for solution)
Velocity of ultrasonic wave in solvent is represented by UO & in solution by Us
Results & discussion
From the following observation table 1,2 & 3 the values of φK(s) φv is higher in
case of acetone water mixture than ethanol water of methanol water mixture due to bulky
electron releasing group present in acetone & also due to difference in functional group
& hydration numbers in case of acetone. The Values of φK(s) in different organic solvent
& water mixture for only 20% is may be due to the fact of polar nature of organic
solvent. The values of φv is in the order as follows,
Acetone-Water >Methanol-water >Ethanol-water at 20% solutions. Thus there is
no regular order of φK(s) & φv values for all the system under investigation. The same
order is observed by (Pankanti S.U. Ph.D. Thesis in Chemistry Marathwada University
Aurangabad (1986) for amino acids, (Narwade etal Acaustica, 82, 1(1996) for substituted
diketones.
3
3
2
2
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Table - 1
Ultrasonic Velocity in Distilled Water
Ultrasonic Frequency = 2 MHZ Temp. 27± 0.1°C
Sr.
No.
Number of
rotation of
Screw
Micrometer
Reading
(mm)
Difference
between
Reading
(mm)
Distance
traveled by
Screw for one
maxima [(D)
mm]
Ultrasonic
Velocity
[(U)msec-1
]
1 5 21.33 - 0.3840 1536
2 10 19.38 1.95 0.3830 1532
3 15 17.44 1.94 0.3820 1528
4 20 15.51 1.93 0.3810 1524
5 25 13.59 1.92 0.3800 1520
6 30 11.69 1.90 0.3790 1516
Average 0.3815 1526
U = 1526 m sec-1
U with standard deviation = 1526 + 0.96 m sec-1
Table - 2 (a)
Ultrasonic Velocity In 20% Ethanol Water Mixture
System - Ligand (L1)
Ultrasonic Frequency = 2 MHz Temp. 27± 0.1°C Molality =0.0072
Sr.
No.
Number of
rotation of
Screw
Micrometer
Reading
(mm)
Difference
between
Reading
(mm)
Distance
traveled by
Screw for one
maxima [(D)
mm]
Ultrasonic
Velocity
[(U)msec-1
]
1 5 19.74 - 0.3780 1512
2 10 17.81 1.93 0.3770 1508
3 15 15.89 1.92 0.3760 1504
4 20 13.98 1.91 0.3750 1500
5 25 12.08 1.90 0.3740 1496
6 30 10.19 1.89 0.3730 1492
Average 0.3755 1502
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Table – 2 (b)
Ultrasonic Velocity In 20% Acetone Water Mixture
System - Ligand (L1)
Ultrasonic Frequency = 2 MHz Temp. 27± 0.1°C Molality =0.0072
Sr.
No.
Number of
rotation of
Screw
Micrometer
Reading
(mm)
Difference
between
Reading
(mm)
Distance
traveled by
Screw for one
maxima [(D)
mm]
Ultrasonic
Velocity
[(U)msec-1
]
1 5 24.35 - 0.3690 1476
2 10 22.40 1.95 0.3680 1472
3 15 20.46 1.94 0.3660 1464
4 20 18.53 1.93 0.3650 1460
5 25 16.61 1.92 0.3640 1456
6 30 14.70 1.91 0.3630 1452
Average 0.3658 1463.3
Table - 2 (c)
Ultrasonic Velocity In 20% Methanol Water Mixture
System - Ligand (L1)
Ultrasonic Frequency = 2 MHz Temp. 27± 0.1°C Molality =0.0072
Sr.
No.
Number of
rotation of
Screw
Micrometer
Reading
(mm)
Difference
between
Reading
(mm)
Distance
traveled by
Screw for one
maxima [(D)
mm]
Ultrasonic
Velocity
[(U)msec-1
]
1 5 23.10 - 0.3870 1548
2 10 21.12 1.98 0.3860 1544
3 15 19.16 1.96 0.3860 1544
4 20 17.22 1.94 0.3850 1540
5 25 15.29 1.93 0.3840 1536
6 30 13.27 1.92 0.3830 1532
Average 0.3851 1546
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Table 3
φK(s) & φv values along with other parameters in 20% of ethanol-water, methanol-water
& acetone-water mixtures of ligand or peptide carnosine (L1)
Ultrasonic frequency: 2MHz Temp 27+0.10c
Parameters Solvent percentage (v/v) 20
Ethanol-water Methanol-water Acetone-water
Molality
Do
do
Uomsec-1
BoX108
Ds
ds
Usmsec-1
BsX108
BodsX108
bsdoX108
φK(s)X104
φv
0.0072
0.3670
1.0461
1468.00
44.3655
0.3755
1.0446
1502.00
42.4
46.2757
44.3546
0.8849
443.24
0.0072
0.3725
1.0258
1490.00
43.90
0.3851
1.0115
1546.00
41.3
44.4048
42.3656
-28.4600
2360.63
0.0072
0.3660
1.0492
1464.00
44.40
0.3658
1.0091
1463.30
46.2
44.8040
48.4730
48.6895
6119.7
References
Kor S.K., Bhatti S.S., “Ind. J.Pure Appl. Phy.,” 7, 784(1969)
Soitkar V.S., Jajoo S.N., “Acoustic Lett.,” 7, 191 (1984)
Satyavati A.V., “Acoustica,” 70, 40 (1984)
Ramchandran K., “Ind. J.Pure. Appl. Phy.,” 6, 75(1968)
Prakash S., Shrivastav S.P., “Ind. J. Chem.,” 3, 489 (1964)
Marks G.W., “J. Acoust. Soc., America,” 38, 327 (1960)
Agrawal S.B., Bhatnagar P.P., “Acoust. J. Phys.,” 31, 567 (1978)
Tabhane V.A., “Acoustic Lett.” 6, 120 (1983)
Tabhane V.A., Ghosh S., Pranjale A.W., “Ind. J. Pure and Appl. Phys.,” 23, 502 (1985)
Prasad N., Prakash O., “Ultrasonics,” 18, 160 (1980)
Kumar A., Prakash S., Singh R., “J. Acoust. Soc., Ind.” 4, 39 (1976)
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Singh S., Prasad N., Prakash O., Chaturvedi C.V., “Ind. J. Chem.” 9, 1138 (1970)
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Gupta A., Mandal A.K., “Ind. J. Pure and Appl. Phy.” 26, 340(1988)
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Institution (b) D1193-70 (1970) Standard Specification for Reagent Water Easto,
Md.;American Society for Testing Materials (c) Reagent Chemicals; Supplement
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Flory P.J., Orwell R.A., “J. Am. Chem. Soc” 86, 3507 (1964)
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Gnananba S. and Rao B.R., “Ind. J. Pure Appl. Phys.” 7, 468 (1969)
Prigogine I., Bellemans A., Englest, “J. Chem. Phys.,” 24, 518 (1956)
Kore.S.K., Batti S.S., “Ind. J. Pure Appl. Phys.”, 7, 784 (1969)
Soitkar V.S., Jajoo S.N, “Acoustic Lett.” 7.191 (1984)
Millerio F.J. Surdo A.L., Shin C, “J. Phys. Chem”, 82, 784 (1978)
Kaulugud M.V., Patil K.J., “Ind J. Pure and appl Phys”, 13,322 (1975)
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Khobragade B.G., “Ph.D Thesis in Chemistry”, Amravati university Amravati (1999)
Pankanti S. U., “Ph.D. Thesis in Chemistry”, Marathwada University, Aurangabad
(1986)
Sawalakhe P.D., Narwade M.N., “Acoustica”, 82, 1 (1996)
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