Experiment No.

2
Factors Affecting Drug Action

Influence of Chemical Structure

Section B3
Objectives

 Todetermine the effect of epinephrine,

dobutamine and terbutaline on the rate of
cardiac contraction

 To determine the effect of epinephrine,

dobutamine and terbutaline on the amplitude
of cardiac contraction
 To determine if there is a significant
difference among the three experimental
values
Hypothesis

Null – there is no significant increase in the rate

and amplitude among the three drugs
Alternative – there is a significant increase in
the rate and amplitude among the three
drugs
Materials

 Kymograph set-up  Surgical thread

 Kymograph paper  Heart clip
 Paste  Dissecting set
 Cotton thread  Tyrode solution
 Surgical gloves  Tuberculin syringe
Materials

 Epinephrine 1:1000 ampule

 Dobutamine 250 mg/ 20 mL vial
 Terbutaline 2.5 mg/ mL
Procedure and Rationale

1. Expose turtle heart.

2. Attach kymograph set-

up through apex of
the heart.
1. Get baseline rate and
amplitude of
contractions for 1
minute.

4. Instill drugs slowly and

directly to heart
muscle (1 ml in 1
minute).
5. Get rate and amplitude for 3
minutes from instillation.

6. Wash turtle heart with tyrode

solution in between drug
administration.

7. Wait for 10 minutes before

administration of next drug.

8. Analyze results statistically and set

p at 0.50. Kymograph
Results
 Rate
section turtle epinephrine Dobutamine terbutaline
After After After
Baseline Infusion Baseline Infusion Baseline Infusion

A 1 41.0 51.0 46.0 49.0 43.0 44.0

2 41.0 46.0 44.0 45.0 43.0 44.0

B 1 50.0 54.0 36.0 38.0 34.0 35.0

2 36.0 54.0 42.0 46.0 36.0 36.0

C 1 54.0 50.0 49.0 47.0 32.0 46.0

2 51.0 52.0 47.0 39.0 47.0 34.0

D 1 32.0 37.3 34.0 32.0 35.0 32.0

2 28.0 33.5 32.0 29.6 28.0 21.0

Mean 41.6 47.2 42.3 40.2 39.3 36.5

Effect on the heart rate when the following drugs are instilled to the heart
muscle.

Effect of Epinephrine on Heart Rate

60.0

50.0
Rate (beats/min)

40.0
baseline
30.0
epinephrine
20.0

10.0

0.0
1 2 3 4 5 6 7 8
Turtle
 Effect of Dobutamine on Heart Rate

60.0
Rate (beats/min)

50.0
40.0
baseline
30.0
Dobutamine
20.0
10.0
0.0
1 2 3 4 5 6 7 8
Turtle
 Effect of Terbutaline on Heart Rate

50.0
45.0
40.0
Rate (beats/min)

35.0
30.0
baseline
25.0
Terbutaline
20.0
15.0
10.0
5.0
0.0
1 2 3 4 5 6 7 8
Turtle
 Amplitude
Section turtle Epinephrine Dobutamine Terbutaline
Baseli
ne After Infusion Baseline After Infusion Baseline After Infusion

A 1 3.0 2.0 0.7 1.1 1.5 1.7

2 0.5 0.7 0.5 0.6 0.4 0.5

B 1 1.1 1.9 1.0 1.4 0.8 0.9

2 1.2 1.3 1.1 1.6 1.0 1.2

C 1 --- 1.1 1.9 1.0 1.2 1.4

2 0.9 1.2 0.5 1.4 --- 1.0

D 1 0.5 6.0 0.5 10.0 1.0 6.0

2 0.6 5.5 0.2 4.0 1.5 6.0

Mean 1.1 2.5 0.8 2.6 1.1 2.3

Effect on the amplitude of the heart when the following drugs are
instilled to the heart muscle

Effect of Epinephrine on Amplitude

3.50
Amplitude (mm)

3.00
2.50
2.00 baseline
1.50 Epinephrine
1.00
0.50
0.00
1 2 3 4 5 6 7 8
Turtle
 Effect of Dobutamine on Amplitude

2.00
Amplitude (mm)

1.50

baseline
1.00
Dobutamine

0.50

0.00
1 2 3 4 5 6 7 8
Turtle
 Effect of Terbutaline on Amplitude

1.80
1.60
Amplitude (mm)

1.40
1.20
1.00 baseline
0.80 Terbutaline
0.60
0.40
0.20
0.00
1 2 3 4 5 6 7 8
Turtle
Expected Result

Rate Epinephrine > Dobutamine > Terbutaline

Amplitude Epinephrine > Dobutamine > Terbutaline

Average of % differences for Rate

60
50
40
30 Series1
20
10
0
1 2 3
Average of % differences for
Amplitude

50
40
30
20 Series1
10
0
-10 1 2 3
Actual Result

Rate Epinephrine > Dobutamine> Terbutaline

Amplitude Dobutamine >Epinephrine > Terbutaline

Discussion
Effects of Sympathomimetic Drugs on
the Heart

• Determined largely by beta-1, lesser

extent beta 2 and alpha receptors are also
involved

• β1 agonist: increase force and rate of

contraction, increase impulse conduction
Epinephrine

 Dihydroxylated (3 & 4)
 catecholamine
 Maximal α and β
activity
Epinephrine
 α1 = α2; β1 = β2
 α1- positive inotropic
response
 β1- positive inotropic,
chronotropic actions on
heart

Can J Physiol Pharmacol 2000 Apr;78(4):267-92

Dobutamine
 Dihydroxylated (3 & 4)
 Catecholamine
 Prominent inotropic

 β1 > β2>>> α
Terbutaline
 Dihydroxylated (3 & 4)
 3 methyl substituents
 β2 >> β1 >>>> α

 CV effect: indirectly 
enhance NE release
after activation of
presynaptic B2
receptors
 Terbutaline
Dobutamine
Percent Difference
SECTION TURTLE RATE (% Difference) AMPLITUDE (% Difference)

Epinephrine Dobutamine Terbutaline Epinephrine Dobutamine Terbutaline

A 1
24.39 6.52 2.33 33.33 57.14 13.33
2
12.20 2.27 2.33 40.00 10.00 12.50
B 1
8.00 5.56 2.94 72.73 40.00 12.50
2
50.00 9.50 0.00 8.30 45.00 20.00
C 1
1.96 0.00 2.13 17.02 117.39 140
2
3.70 20.41 6.25 120 26.32 18.03
D 1
16.56 5.88 8.57 20.00 100.00 40.00
2
19.63 7.50 25.00 0.00 100.00 60.00
Statistics
ANOVA (analysis of variance)
ANOVA-Single Factor (p value=0.05)

 p-value of 0.102 is greater than

0.05 thus there is no significant Rate 0.102
increase in the rate of the heart
when epinephrine, dobutamine
and terbutaline was instilled
slowly to the heart muscle.
 On the other hand, the p-value Amplitude 0.453
of the amplitude is 0.453 that is
greater than 0.05 thus there is
no significant increase in the
amplitude when epinephrine,
dobutamine and terbutaline was
instilled slowly to the heart
muscle.
Sources of Errors

 Faulty kymograph
 Animal preparation
 Efficiency of Heart Washing
Recommendation

 Use of a bigger animal

 Use of different animal for the different drugs
 Better monitoring of the results
 More efficient animal preparation
Conclusion

 There was an increase in rate and amplitude

upon administering of the three drugs on the
cardiac contraction of the turtle.
 However, there was no significant difference
among the three drugs in the rate and
amplitude of cardiac contraction of the
turtles’ hearts.
Thank you!

MEMBERS: B3
dela Pena, Jean Joan; dela Pena, Nathaniel; Dela rosa,
Haziel; Desquitado, Jill; Detera, Carla; Diansuy,
Wilfredo; Dimal, Gianina; Dimalala, Gretel; Dimaunahan,
Eric; Din, Marian Bernadette; Djajakasuma, Angela;
Doma, Lorraine; Dumantay, Mae Dorothy;
Domingo,Dumont