Pharmacology 2

Diuretics lab #2
Shevine minto 1901751
Dr. Brown
September 15,2021
Tuesday 8-11am class
 PRACTICAL #2 – DIURETICS

INTRODUCTION
A diuretic is an agent which increases the volume of urine formed by increasing the mobilization
of extracellular fluid (ECF), resulting in the loss of ions (sodium, potassium, chloride, etc) and
water. There are many different agents that act by various mechanisms to produce diuresis.
The following experiment was carried out using an anaesthetized dog (IV 30 mg of
pentobarbitone sodium/kg). The weight of the dog was 1600g.

PREPARATION:
1. The trachea, carotid artery, and femoral vein were cannulated.
2. A midline incision was made from the umbilicus to the symphysis pubis.
3. Holding the fundus of the urinary bladder it was pulled through the incision and
downward to expose the posterior surface.
4. The ureters were located as they enter the bladder without grasping them with forceps.
5. A polythene tube was inserted into the ureters and tied.
6. The number of drops was recorded on a kymograph.
7. The number of drops per minute from the tracing were noted.
8. Urine was also collected in a measuring cylinder and the e volume per minute over a
given period of time (10-15 mins.) after the administration of each drug.

PROCEDURE:
1. A 5-minutes control record was secured.
2. 1.0 ml phenolsulphonphthlein (P.S.P.) (dye) was injected intramuscularly.
3. Urine was collected in a beaker.
4. The time was recorded from the injection of the dye to the first appearance of a purple
colour in the beaker.
5. 0.5 ml of 10% urea per kg was injected.
6. 0.5 ml of 50% glucose per kg was injected.
7. 0.5 ml of 50% sucrose per kg was injected.
8. 0.5 ml of 0.5 mg/kg of frusemide (Lasix) was injected.
9. 0.5 ml of 0.5 mg/kg of Mersalyl was injected very slowly.
10. 0.5 ml of 10 mg/kg of HgCl2. was injected very slowly.
Table 1. Drops/Volume of Urine/Unit Time

VOLUME/MINS
METHOD COMMENTS
0 2 4 6 8 10
1. Collect urine for 10 0.5 is a normal volume of urine ,
mins as control 0. amounts are constant because
0 0.5 0.5 0.5 0.5 no diuretic is administered to
5
increase the volume
2. Inject 0.5 ml 10% The volume rises to a maximum
urea/kg of 0.8 and then begins to
0. decrease again. The duration of
0 0.7 0.8 0.6 0.5
5 action lasts for 8 minutes before
the volume begins to decrease
again
3. Inject 0.5 ml 50% The onset of glucose was slow
glucose/kg compared to urea. Urine output
0.
0 0.5 0.8 0.7 0.5 reached a maximum of 0.8 and
5
it had a short duration of action
of about 4 minutes
4. Inject slowly 1ml of The onset of mersalyl was fast,
0.05 ml/kg mersalyl it continues to increase even at
0. 10 minutes which means it has a
0 0.7 0.8 0.85 0.9
5 very long duration of action.
Mersalyl is more effective as a
diuretic than glucose and urea
5. Inject 1ml of 0.5 mg/kg Lasix had an extremely fast
frusemide (Lasix) onset, yielding the highest
volumes of urine. The volume of
urine however fluctuates
1.
0 1.3 1.2 1.0 1.2 regularly. It had a modal
2
volume of 1.2. it increased in the
first 4 minutes , decreased for
the next 4 minutes then
increased again to 1.2
HgCl2 had no diuretic effect on
6. Inject very slowly 1ml the dog and had the same
of 10 mg/kg HgCl2 results as the control
0.
0 0.5 0.5 0.5 0.5 experiment
5

NOTE:
- Except for P.S.P all drugs were given intravenously.
 - Control rates were done between additions of each drug.
- The normal urine production for dogs is 30mL/Kg/24hrs.

Activity

1. Complete the table by commenting on the findings. (6

marks)

2. Calculate how much of the following substances were injected into the dog. One calculation
to be shown and units to be indicated.
(5 marks)

Substance injected Amount

Urea 0.5ml 10% urea/kg

1600g=1.6kg
10g=100ml
X=0.5ml
X=0.05g
0.05/kg x 1.6kg
0.08g

Glucose 0.4g

Mersalyl 0.5/ml of 0.5/kg

0.5=1kg
X=1.6kg
0.5mg x 1.6kg=0.8mg
0.5mg=1ml
0.8=x
X=1.6ml

Frusemide 1.6ml

HgCl2 32ml

3. Plot a graph of number of drops/2 mins against time. (10

marks)
 NB – Graph papers are needed.

4. Based on the results comment on the onset and duration of diuresis of urea, glucose and
frusemide. (6
marks)

Agent Onset of action Duration of Diuresis

Urea Fast onset Short duration

Glucose Slow onset Short duration

Frusemide fast onset Long duration

5. Explain the mechanism of action by which these agents produce a fall in blood pressure.
(6
marks
)

Diuretics, sometimes called water pills, help rid your body of salt (sodium) and water. Most of
these medicines help your kidneys release more sodium into your urine. The sodium helps
remove water from your blood, decreasing the amount of fluid flowing through your veins and
arteries. This reduces blood pressure.

6. Give three (3) differences between furosemide (Lasix) and hydrochlorothiazide (HCTZ)
 (3
marks
)

Difference Furosemide HCTZ

1. Acts on the ascending loop Acts on the distal

convoluted tube

2. Blocks Na K Cl symport Blocks Na Ca symport

3. Is a loop diuretic Is a thiazide diuretic

7. Describe the two ways that potassium-sparing diuretics work? (4

marks)
a. bind to aldosterone receptors to reduce sodium and water retention and decrease
potassium excretion(Sodium channel blocker)
b. inhibit Na+ re-entry through the ion channel(Aldosterone antagonists)

8. Patients taking potassium-sparing diuretics are most at risk for which electrolyte imbalance?
(1
mark)
potassium

Table 2.

PROCEDURE OBSERVATION EXPLANATION & COMMENT

Inject 1.0 ml of Time of Time of The experiment can continue.

PSP Injection appearance in The normal urine output for the
urine dog is 30ml/24hr. The output
from the experiment was
 10:00 a.m. 10.20 a.m. 45ml/24hr, even though this is
higher it is not too far off and it
can be concluded that the kidneys
are functioning normally

Activity

1. Calculate the rate of urine production in the experiment’s dog. Show working. (2
marks)
1ml/1.6kg/20 mins
20mins=1/3hr
0.625ml/kg/20mins
0.625/kg/0.3hr
=0.625 x 3 x 24hr
=45ml/kg/24hrs

30ml/1.6kg/24hr

Assuming 1ml of urine in 20mins

=48ml
2. Complete the table by commenting on this value and state whether the experiment may
continue. (2
marks)

TOTAL = 45 MARKS