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Online International Interdisciplinary Research Journal, {Bi-Monthly}, Volume-II, Issue-II, Mar-Apr2012

w w w . o i i r j . o r g I S S N 2 2 4 9 - 9 5 9 8

Page 281

“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



82

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



Page 283

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



Page 284

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;



Page 285

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



Page 286

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



Page 287

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



Page 288

Table – 2 (b)

Ultrasonic Velocity In 20% Acetone Water Mixture



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



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



Page 289

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

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Soitkar V.S., Jajoo S.N., “Acoustic Lett.,” 7, 191 (1984)

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Page 290

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