In vitro identification of unknown compounds A, B and C in a
pharmacology practical.
Tymo Zwakenberg S4326253
Group 16
Date of experiments: 26-10-2022, 3-11-2022 and 22-11-2022
Date of submission:
Dr. H.A. Baarsma
Ing. A.B. Zuidhof
1
,Table of contents
- Introduction and theory
o Objective
o Theoretical principles
o Ileum
o Aorta
o Atrium
- Materials and methods
o Organ baths
o Execution and calculation examples
- Results
o Ileum
▪ Compound A
▪ Metacholine response curve
▪ Compound A + metacholine
▪ Compound B
▪ Compound C
o Aorta
▪ Compound A, B & C
▪ Phenylephrine loading dose curve
▪ Compound A + phenylephrine
▪ Histamine loading dose curve
▪ Compound B + histamine
▪ Phenylephrine loading dose curve
▪ Compound C + phenylephrine
▪ Histamine loading dose curve
▪ Compound C + histamine
o Atrium
▪ Histamine loading dose curve
▪ Compound B + histamine
▪ Compound C
▪ Compound C + mepyramine
▪ Compound C + ketanserin
- Discussion and conclusion
o Experimental day 1
▪ Inclusion/exclusion of possibilities
o Experimental day 2
▪ Inclusion/exclusion of possibilities
o Experimental day 3
▪ Inclusion/exclusion of possibilities
o In vivo
o Compound A
o Compound B
o Compound C
- References
2
,- Appendices
o Appendix A: Ileum
▪ Histamine dose response curves
• Appendix figure 1
• Appendix figure 2
• Appendix figure 3
• Appendix figure 4
▪ Appendix calculations
▪ Table of raw data
o Appendix B: Aorta
▪ Histamine dose response curves
• Appendix figure 5
• Appendix figure 6
• Appendix figure 7
• Appendix figure 8
▪ Appendix calculations
▪ Table of raw data
o Appendix C: Atrium
▪ Histamine dose response curves
• Appendix figure 9
▪ Appendix calculations
▪ Table of raw data
3
, Introduction and theory
Objective
The execution of the pharmacology practical designed like this should provide in the development of
pharmacological principles existing of independently setting up experimental ideas, researching on
animal organ material in organ baths, organizing group coöperation, working with agonistic and
antagonistic compounds and ultimately indentifying unknown compounds. The execution of the
practical should setup experimental executioners for further pharmacological experiments and
provide the basis which is necessary in that regard. Each experimental day provides the chance to
get closer to the identity of the unknown compounds. However, although the identification of the
compounds is a central part of the practical, the reasoning and thought processes that will be the
backbone of the results and discussion are at least – if not even more important than the final
compound identification itself. Since the practical should provide in the knownledge of
pharmacological aspects and not as much about the characteristics of the unknown compounds, the
reasoning and thought processes that will be discussed should be considered heavily in order to get
a better grip on this pharmacology practical and possible upcoming pharmacological practicals.
Theoretical principles
Before the practicals can be attended, knowlegde about the known compounds, experimental setup
of the organ baths, possible targetable receptors, characteristics of the particular organ and the goal
of the day should be sufficient. Preparation and the knowledge about the stated components are
necessary in order to succeed in the execution of the practical. This and the fact that animal organ
material is used should stimulate personal responsibility to be prepared and work accordingly. The
information that was used throughout the practicals will be discussed along the results and
discussion section in order to provide the explanations.
All the substances that are provided and used throughout the practical, whether it is a known
compound or an unknown compound resemble an agonistic or antagonistic influence on the organ
samples. An agonistic influence would result in the contraction of the muscle that is present in that
specific organ. On the other hand should the antagonistic influence not show contracting
characteristics but relaxational characteristics, this is mostly observable after a contraction has
occured. However, the antagonistic influence can also be indicated through the occurance of a right
shift in agonistic concentration in order to exert contraction. To clarify this the stated effects of an
agonist and antagonist are represented in figure 1 & 2.
Figure 1: Characteristic concentration-effect curve after the addition of increasing agonist
concentrations [4].
4
pharmacology practical.
Tymo Zwakenberg S4326253
Group 16
Date of experiments: 26-10-2022, 3-11-2022 and 22-11-2022
Date of submission:
Dr. H.A. Baarsma
Ing. A.B. Zuidhof
1
,Table of contents
- Introduction and theory
o Objective
o Theoretical principles
o Ileum
o Aorta
o Atrium
- Materials and methods
o Organ baths
o Execution and calculation examples
- Results
o Ileum
▪ Compound A
▪ Metacholine response curve
▪ Compound A + metacholine
▪ Compound B
▪ Compound C
o Aorta
▪ Compound A, B & C
▪ Phenylephrine loading dose curve
▪ Compound A + phenylephrine
▪ Histamine loading dose curve
▪ Compound B + histamine
▪ Phenylephrine loading dose curve
▪ Compound C + phenylephrine
▪ Histamine loading dose curve
▪ Compound C + histamine
o Atrium
▪ Histamine loading dose curve
▪ Compound B + histamine
▪ Compound C
▪ Compound C + mepyramine
▪ Compound C + ketanserin
- Discussion and conclusion
o Experimental day 1
▪ Inclusion/exclusion of possibilities
o Experimental day 2
▪ Inclusion/exclusion of possibilities
o Experimental day 3
▪ Inclusion/exclusion of possibilities
o In vivo
o Compound A
o Compound B
o Compound C
- References
2
,- Appendices
o Appendix A: Ileum
▪ Histamine dose response curves
• Appendix figure 1
• Appendix figure 2
• Appendix figure 3
• Appendix figure 4
▪ Appendix calculations
▪ Table of raw data
o Appendix B: Aorta
▪ Histamine dose response curves
• Appendix figure 5
• Appendix figure 6
• Appendix figure 7
• Appendix figure 8
▪ Appendix calculations
▪ Table of raw data
o Appendix C: Atrium
▪ Histamine dose response curves
• Appendix figure 9
▪ Appendix calculations
▪ Table of raw data
3
, Introduction and theory
Objective
The execution of the pharmacology practical designed like this should provide in the development of
pharmacological principles existing of independently setting up experimental ideas, researching on
animal organ material in organ baths, organizing group coöperation, working with agonistic and
antagonistic compounds and ultimately indentifying unknown compounds. The execution of the
practical should setup experimental executioners for further pharmacological experiments and
provide the basis which is necessary in that regard. Each experimental day provides the chance to
get closer to the identity of the unknown compounds. However, although the identification of the
compounds is a central part of the practical, the reasoning and thought processes that will be the
backbone of the results and discussion are at least – if not even more important than the final
compound identification itself. Since the practical should provide in the knownledge of
pharmacological aspects and not as much about the characteristics of the unknown compounds, the
reasoning and thought processes that will be discussed should be considered heavily in order to get
a better grip on this pharmacology practical and possible upcoming pharmacological practicals.
Theoretical principles
Before the practicals can be attended, knowlegde about the known compounds, experimental setup
of the organ baths, possible targetable receptors, characteristics of the particular organ and the goal
of the day should be sufficient. Preparation and the knowledge about the stated components are
necessary in order to succeed in the execution of the practical. This and the fact that animal organ
material is used should stimulate personal responsibility to be prepared and work accordingly. The
information that was used throughout the practicals will be discussed along the results and
discussion section in order to provide the explanations.
All the substances that are provided and used throughout the practical, whether it is a known
compound or an unknown compound resemble an agonistic or antagonistic influence on the organ
samples. An agonistic influence would result in the contraction of the muscle that is present in that
specific organ. On the other hand should the antagonistic influence not show contracting
characteristics but relaxational characteristics, this is mostly observable after a contraction has
occured. However, the antagonistic influence can also be indicated through the occurance of a right
shift in agonistic concentration in order to exert contraction. To clarify this the stated effects of an
agonist and antagonist are represented in figure 1 & 2.
Figure 1: Characteristic concentration-effect curve after the addition of increasing agonist
concentrations [4].
4
