STUDIA UNIVERSITATIS BABES-BOLYAI, PHYSICA, SPECIAL ISSUE, 2003
HEMOGLOBIN SOLUTIONS IN ULTRASONIC FIELD
Alina Rapa1 Servilia Oancea2 Dana Dorohoi3
1.C.A.Johnson High School Columbia SC (USA), email: arapa@richlandone.org
2.Univ.Of Agronomy and Veterinary Medicine, Iasi,
Romania, e-mail:lioancea@univagro-iasi.ro
3.“Al.I.Cuza “ Univ, Faculty of Physics, Iasi,
Romania
Abstract
High-frequency acoustic waves interact with living systems. Knowledge
on the ultrasound propagation process through mammalian tissues is of
the great interest for application in medicine, farmacy and agriculture. In
what hemoglobin is concerned, hemoglobin as well as the hematocrit and
erythrocyte counting is of a great importance for screening and diagnosis
related to anemia. That is the reason for us to study some experimental
aspects linked with behavior of hemoglobin in ultrasonic field.
Ultrasound velocity was interferometrically determined. Adiabatic
coefficient of compresibility has been estimated from ultrasound velocity
and the solution density. At 200C, the ultrasound velocity in solutions
increases with the concentration of cow blood hemoglobin in water
solution, ranging between 1520-1620 m/s for concentration between 540mg/cm3. For human hemoglobin ultrasound velocity in water solutions
at 200C ranged between 1492-1540 m/s, also increasing with
concentration.
Concomitantly, an increase of absorbtion coefficient with concentration
was determined. The water absorbtion was substracted from the total
absorbtion to obtain information on protein absorbtion.
1. Introduction
Knowledge on the ultrasound propagation process through mammalian
tissues is of the great interest for application in medicine, pharmacy and agriculture
[1], [2]. In what hemoglobin is concerned, hemoglobin as well as the hematocrit
and erythrocyte counting is of a great importance for screening and diagnosis
related to anaemia [3]. The development of atherosclerosis and the failure of
vascular grafts are often associated with specific characteristic of local
hemodynamics. Clinically, ultrasound velocity measurement can be used to
estimate wall shear rate in vivo [4].
Investigation of blood properties from various species has attracted
considerable interest and comparative studies have yielded interesting aspects
about the behaviour of blood [5]. [6].
ALINA RAPA SERVILIA OANCEA DANA DOROHOI
2. Materials and methods
We obtained the hemoglobin solutions by treating washed, packed red
blood cell from cow blood with toluene to liberate the hemoglobin. By
centrifugation, the stroma associated with toluene is separated from the heavier
hemoglobin solution. Then a hemoglobin solution can be obtained by adding
distilled water. The action of osmotic forces on the red blood cells causes
hemolysis and free hemoglobin is distributed through the solution.
We didn’t make spectral analysis in order to estimate the portion of
oxihemoglobin or methemoglobin in the studied solutions, because the ultrasound
absorption by these derivatives is substantially the same [7]. Ultrasound velocity
was interferometrically determined using an ultrasound pulse method. A laboratory
device designed and assembled in our laboratory was used to measure wave
velocity and to test behaviour of ultrasonic wave in a hemoglobin sample (Fig.1).
An ultrasonic oscillator gives a signal that propagates through the hemoglobin
solution. The direct signal and the one propagated through the hemoglobin solution
are simultaneously visualized on the oscilloscope screen [8]. Velocities were
measured, for different US frequencies, using the value of propagation time
through solution:
t  n
where: -n represents the number of divisions that separates the two signals on the
screen
-  is the time base of oscilloscope
Fig.1 Bloch schema for determination of the velocity and absorption coefficient of
ultrasound in hemoglobin solutions
HEMOGLOBIN SOLUTIONS IN ULTRASONIC FIELD
The ultrasound velocity is:
c
L
t
with L-the length of the propagation cell. Ultrasound absorption has been estimated
in terms of absorption (  ) per wavelength (  ),  in nepers. Absorption
coefficient  has been measured from the intensity I of two signals at the distance
L related to the oscillator:
I1
I2

L2  L1
ln
The wavelength of US has been calculated with formula:

c

where are ultrasound velocity and  the ultrasound frequency.
3.Results and discussion
The results of the measurements for US velocity are presented in Table 1.
Table 1. US velocity in hemoglobin solution.
1
2
4
7
5
1520.0
1523.0
1524.0
1524.3
10
1530.1
1530.4
1530.8
1531.2
15
1541.5
1541.5
1541.7
1542.0
20
1562.4
1562.5
1563.1
1563.3
25
1581.7
1582.2
1582.4
1582.7
30
1607.7
1608.2
1608.7
1609.3
40
1619.1
1619.3
1620.1
1620.8
The dependence of US velocity by Hb concentration (mg/cm3) for 1MHz is
given in the Fig.2
velocity
US velocity versus Hb concentration
1640
1620
1600
1580
1560
1540
1520
1500
0
10
20
30
40
50
Hb concentration
Fig.2. US velocity versus Hb concentration for 1 MHz
ALINA RAPA SERVILIA OANCEA DANA DOROHOI
The dependence of the US velocity from frequency of the ultrasonic wave
is given in Fig.3
US velocity versus frequency for 5 Hb
concentration
1525
velocity
1524
1523
1522
1521
1520
0
2
4
6
8
fequency(MHz)
Fig.3 US velocity as a function of frequency for 5 mg/cm3 concentration
For 40mg/cm3 concentration the dependence US velocity from frequency is
given in Fig.4
US velocity versus frequency for 40 Hb
concentration
1621
velocity
1620.5
1620
1619.5
1619
0
1
2
3
4
5
6
7
8
frequency(MHz)
Fig.4. US velocity as a function of frequency for 40mg/cm3 concentration
The results of the measurements for absorption are presented in Table 2
HEMOGLOBIN SOLUTIONS IN ULTRASONIC FIELD
Table 2.US absorption (  )*103(nepers) in Hb solutions
1
0.64
0.74
1.82
3.00
3.80
5.80
7.10
5
10
15
20
25
30
40
2
0.71
0.78
2.10
3.10
4.00
6.00
7.20
4
0.80
0.81
2.20
3.21
4.10
6.10
7.50
7
1.00
1.20
2.50
3.35
4.20
6.30
7.76
The US absorption versus Hb concentration for 1 MHz is given in Fig.5
absorbtion(nepers)
US absorbtion versus Hb concentration
8
7
6
5
4
3
2
1
0
0
10
20
30
40
50
concentration
Fig.5 US absorption versus Hb concentration for 1 MHz
4.Conclusions
1. Ultrasound velocity in hemoglobin solutions increases to the concentration
of hemoglobin solution, these increases not being affected by ultrasound
frequency.
2. Ultrasound velocity in hemoglobin solutions increases to the frequency but
this dependence is different for different concentrations.
3. US absorption in hemoglobin solutions also increases to the concentration
of hemoglobin solutions.
Further investigations will be developed in order to evaluate the US
velocity and US absorption in hemoglobin solutions of blood for other animals.
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ALINA RAPA SERVILIA OANCEA DANA DOROHOI
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