Prueba 9 2019, questions and answers
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Prueba 9 2019, questions and answers
Farmaceutische Analyse C (Rijksuniversiteit Groningen)
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Prueba 9 2019, questions and answers
Farmaceutische Analyse C (Rijksuniversiteit Groningen)
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Solutions: Mock exam for practice
Question 1
The pKa of an alkaline group in a drug molecule is 10. The partition coefficient P of the
uncharged base is 25. The phase ratio V is 5. What is the fraction of the non-protonated, neutral
ase in the organic phase (forg) at,
(a) pH= 11 and what is forg at, (b) pH= 8?
What is forg if you perform 5 extractions at (c) pH=11 or at, (d) pH=8 in succession?
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Solution
SOWISO 2020 1
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Question 2
Describe the principle of a Fluorescence Polarization assay. How is this principle used to determine
a drug in complex biological samples? Is this a homogeneous or a heterogeneous assay? Please
justify your answer. Is this a competitive or non-competitive assay? Please justify your answer.
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SOWISO 2020 2
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Solution
A fluorescence polarization assay is based on the interaction between a low molecular mass drug
and a high molecular mass antibody or receptor. Furthermore, a fluorescently labeled drug
conjugate is required, which binds the antibody or receptor as much as the free drug in the
sample. The measurement principle is that a large molecule, such as an antibody, moves more
slowly than the drug conjugate. When the sample is i
adiated with polarized light, the unbound
drug conjugate contributes a lot to depolarization, because it moves between the moment of
excitation and light emission (fluorescence) and thus emits the originally polarized light again
on different planes. If the drug conjugate is bound to an antibody or receptor, depolarization
is much less, because the large molecule moves much slower and therefore emits most of the
original light on the same plane again.
For the assay, a fixed amount of fluorescently labeled drug conjugate is added to the sample as
well as a fixed amount