Practical Work 2

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Practical work 2

DIALYSIS

A. Theoretical explanations
B. Quantitative evaluation of dialysis by conductivity
C. Experimental technique
D. Medical application: artificial kidney

A. Theoretical explanations
Some membranes in the body are selectively permeable because they allow the diffusion
(passive transport) of water, salts, glucose, urea and other small organic molecules, but they
block the normal passage of larger macromolecules such as hemoglobin, globulin, albumin and
other larger molecules of protein. In this case the passage of solute through the membrane is
called dialysis.
Thus, for a solution containing several types of molecules, dialysis is a separation process by a
permeable membrane through which will pass only the particles with a diameter smaller than the
diameter of the pores of the membrane. Colloidal particles (such as protein) having a larger
diameter than the pore cannot pass.

Selective mebrane

dyalisis

 
Isotonic solution Excess of electrolytes and colloids

 - Solvent molecule
 - Solute molecule
 - Colloidal molecule
Fig.1. Dialysis
B. Quantitative evaluation of dialysis by conductivity

Conductometry represents a series of methods that measure different electrical parameters


(resistance, conductivity etc) for an electric conductor.
Conductors may be classified in:
first order conductors – for which the electric current travels by the help of electrons as
movable charged particles (for example, metals: copper, aluminum, silver) they are also
called metallic conductors,
second order conductors – the charged particles are represented by the negative anions
and positive cations contained in an electrolytic solution, they are also called electrolytic
conductors.

If we apply a potential difference strong enough between two metallic plates (made of platinum,
for example), submerged in an ionic solution, between the two electrodes appears an electric
current. The circulation of the current is conditioned by the presence of the ions in the solution.
Some physical notions can characterize the passage of the charged particles in the second order
conductors:
Electric resistance of a conductor (R):

(1)
[R] SI = Ω (ohm)
U = potential difference, V (volts);
I = electric current, A (ampere)
Conductance G is the reverse of resistance:

(2)
[G]SI = S (siemens)

The electric resistance of a metallic conductor with a section (S) and a length (L) is determined
by the equation:

(3),
with ρ, the resistivity value for a metal. This resistivity depends on the metal kind and for a
specific metal on the temperature.
For electrolytes solutions the resistance of a portion of electrolyte of parallelepiped shape
situated between the two electrodes has the same expression as that of a metallic conductor (3)
We can measure the resistance (R) of electrolyte solution by a conductivity cell made of two
plane platinum electrodes (covered with a thin layer of powdery platinum, placed parallel one to
another. The length (L) of the conductive solution corresponds to the distance between the
electrodes and the section (S) corresponds to the area of the electrode. We can measure the
resistance R of the solution volume contained between the two electrodes (see the image of the
conductivity cell.

platinum wires

Electrodes (plates
of platinum
covered with
powdery platinum)

clamp

Fig.2 Image of the conductivity cell

Conductivity (λ) is the physical property that allows us to evaluate the concentration of ions in a
solution. It depends not on the system of units; it depends only on the characteristics of the
solution:
ions concentration
nature of ions
temperature of the solution

Conductivity (λ) is the reverse of resistivity:

Then:

The ratio L / S is characteristic for each conductivity cell (cell constant – C, and can be
determined by measuring the resistance of a standard solution)
Then:

from were λ results from the following equation:

If the characteristics of the cell (L and S, or better their ratio C) are known, we can calculate with
relation (7) the values of λ.
C. Experimental technique

A practical possibility for measuring R is to use a method derived from Wheatstone ‘s bridge
(the method that uses Wheatstone’s bridge is dedicated to the measure of the unknown resistance
for metallic conductors).

R2
I2 R3
A C
I1
R1 G
Rx

Fig.3 Wheatstone’s Bridge

The mounting presented in figure 4 bears the name of Kohlrausch’s bridge. This electric device
is supplied in AC (alternative current) under frequencies sufficiently high (at 1000Hz) which
prevent any risk of electrolysis. When the balance of the bridge is reached the instrument zero
(the red LEDs-light emitted diode- are off) gives us the value of resistance. The scale of the
device is graduated in ohm.
Zero conductivity cell
instrument
with LEDs R3
R1

L1 L2

R2 Rx

R1, R2, R3 known values of resistance


Rx unknown resistance

Fig. 4 Kohlrausch’s bridge


As mentioned above, dialysis is a phenomenon of separation by a selective permeability
membrane with the diffusion of several types of molecules having a smaller diameter than the
diameter of pores of membranes. Colloidal particles (like proteins) with a larger diameter than
this cannot pass.
The experimental technique allow the practical study of the phenomenon of dialysis using a
simplified device that model the function of the artificial kidney and evaluate the effectiveness of
dialysis by the conductivity measurement.
The device used has the following components:
1. a compartment representing the inside of the renal glomerulus. This compartment is
filled with a solution which has an excess of electrolytes. It represents the blood with an
excess of metabolites and toxic products (urea) ;
2. a cellophane membrane (with selective permeability) which allows the passage of the
molecules of electrolytes with the diameter smaller then the diameter of its pores ;
3. a compartment representing the outside (bath of dialysis) which is filled with the same
quantity of an isotonic solution (isotonic with the normal blood)

Experiment
Due to the selective permeability of the cellophane membrane a phenomenon of dialysis occurs
from the first compartment to the second. The ionic particles accumulate in the second
compartment thus, conductivity will grow. We figure out this phenomenon by measuring the
electrical resistance (R) of the isotonic solution by conductometry.
In our practical work we will study the variation of resistance (R) and conductivity (λ) during
time:
1. Pour in compartment 1 of the experimental device a quantity of solution with urea excess.
The level of the liquid will slightly exceed half the height of the cellophane membrane.
2. Pour the same quantity of isotonic solution in compartment 2.
3. Note the time.
4. Switch on the device – Kohlrausch’s bridge.
5. With the assist of a pipette take 5 – 10 mL of isotonic solution from the bottle and fill the
conductimetric cell of Kohlrausch’s bridge.
6. Slowly turn the central button from 0 (zero). The LEDs start to extinguish. The bridge is
balanced when a greatest numbers of LED are extinct.
7. Note the value of R0 expressed in ohms.
8. Empty the conductimetric cell content by pressing the clamp and wash the cell (the
interior!) with distilled water.
9. After five minutes from the initial timing repeat the stages 5 to 8 with the liquid from the
compartment 2. You will obtain value R1.
10. Repeat the measurements each 5 minutes during one hour. Note the values. Wash the cell
after each determination.
11. At the end for calculating the corresponding values of conductivity (λ) it is necessary to
calculate the constant C. For doing this we will measure the value of electrical resistance
(RKCl) for a standard solution (KCl, 0.1 N) for which it is known the value of conductivity
(λKCl) of a specific temperature (the dependence conductivity over temperature are given
in the table). Repeat steps 5-9 with the standard solution of KCl.
12. Calculate C with the equation :
13. Calculate values for λ.

Resistance and conductivity of an electrolytic solution are linearly related to the variation in time
of the concentrations of ions in solution. Using the values of R and λ draw the curve on a graph
paper:

R (Ω) Ω-1 m-1)

Time
(min)

Fig. 5 Experimental device


E. Medical application
Artificial kidney:
The properties of the cellophane membrane to separate colloids (such as proteins) from
other diffusible molecules (like ions or smaller molecules) were found by Professor R.
Brinkman. W. Kolff was the first who applied this property in a human condition: renal failure.
When kidney failure occurs, the kidneys cannot remove substances from the blood and these
substances accumulate and increase their plasma concentration. Some of these substances are
highly toxic to human health, such as urea (compound result of cellular metabolism). Even water
and other ions (normally eliminated through urine) may affect the function of some cells (for ex.,
a high concentration of K can influence the appearance of the action potentials in the heart,
leading to ventricular fibrillation and under this circumstances blood cannot leave the heart in
order to irrigate the tissues and organs. That is why in such a condition we need a device to
replace the structure and function of kidney. Hemodialysis is used to eliminate small molecules
from the patient’s blood and this happens for all the molecules that can pass through the pores of
a cellophane membrane (such as water, ions, urea, glucose, etc.), but maintaining in the blood
proteins and cells.

Fig.6 Artificial kidney

The cellophane membrane is used to replace the normal glomerulus membrane due to its
structural characteristics (table 1)
Table 1 Glomerulus membrane vs. artificial membrane

Cellophane
glomerulus membrane
Diameter for pores 30 - 45 Å 30 Å

length pores 1Å 80 Å

Total membrane surface 1500 - 7500 cm2 22000 cm2

As we can see, the diameter of the cellophane pores is similar to glomerulus membrane
and it is possible for the small molecules to pass. The pore length is greater for the cellophane
than the renal glomerulus, and in hemodialysis condition the pores of the cellophane can be
quickly blocked. This inconvenient is compensating by the wider surface of the cellophane
membrane.
In principle, the artificial kidney (fig.6) is composed of a dialyser that contains a
cellophane membrane (the total surface is at least 1 m2). Patient's blood is taken from an artery
and flows into the dialyser. In counter-flow with the blood, an isotonic solution with normal
blood flows. Hemodialysis lasts for 2 to 3 hours. The efficiency of dialysis can be measured by
conductivity of the isotonic solution at certain time intervals. Finally, blood is collected and
enters the patient's body through a vein. In any artificial kidney, there must be respected the
following conditions:
- blood must be kept fluid by adding an anticoagulant substance (heparin);
- all small clots, eventually formed, must be retained by a filter because due to a clot
that enter into a vein circulation embolism may appear;
- bubbles of air must be removed by special filters, to prevent air not to enter into the
vein (this may also produce embolism);
- the amount of blood that circulates outside the patient’s body should be as small as
possible.
Other types of dialysis:
cerebral microdialysis – represents a technique for
measuring the concentration of analytes from extracellular
cerebral fluid. Analytes may include endogenous molecules
(e.g. neurotransmitter, hormones, glucose, etc.) or exogenous
compounds (e.g. pharmaceuticals) to determine their
concentration within the body. The microdialysis technique
requires the insertion of a small microdialysis catheter (also
referred to as microdialysis probe) into the tissue of interest.
peritoneal dialysis (fig.7) - The process uses the
patient's peritoneum as a membrane across which fluids and
dissolved substances and other small molecules are exchanged
from the blood. Fluid is introduced through a permanent tube in
the abdomen and flushed out either every night while the patient
sleeps (automatic peritoneal dialysis) or via regular exchanges
throughout the day (continuous ambulatory peritoneal dialysis).
Fig.7 Peritoneal dialysis

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