Advances in Biopharmaceutics and

Pharmacokinetics

Dr. Muzaffar Abbas

Measurement of Drug Concentrations
Because drug concentrations are an important
element in determining individual or population
pharmacokinetics, drug concentrations are measured
in biologic samples, such as milk, saliva, plasma, and
urine.
Sensitive, accurate, and precise analytical methods
are available for the direct measurement of drugs in
biologic matrices.
Such measurements are generally validated so that
accurate information is generated for
pharmacokinetic and clinical monitoring.
In general, chromatographic methods are most
frequently employed for drug concentration
measurement, because chromatography separates the
drug from other related materials that may cause
assay interference.
Drug Concentrations in Blood, Plasma, or Serum
Measurement of drug concentration (levels) in the
blood, serum, or plasma is the most direct approach
to assessing the pharmacokinetics of the drug in the
body.
Whole blood contains cellular elements including
red blood cells, white blood cells, platelets, and
various other proteins, such as albumin and globulins.
In general, serum or plasma is most commonly used
for drug measurement.
To obtain serum, whole blood is allowed to clot and
the serum is collected from the supernatant after
centrifugation.
 Plasma is obtained from the supernatant of
centrifuged whole blood to which an
anticoagulant, such as heparin, has been added.
 Therefore, the protein content of serum and
plasma is not the same. Plasma perfuses all the
tissues of the body, including the cellular elements
in the blood.
 Assuming that a drug in the plasma is in dynamic
equilibrium with the tissues, then changes in the
drug concentration in plasma will reflect changes
in tissue drug concentrations.
 Serum is the liquid that remains after the blood has clotted.
 Plasma is the liquid that remains when clotting is prevented with the addition of an
anticoagulant.
Plasma Level–Time Curve
The plasma level–time curve is generated by
obtaining the drug concentration in plasma samples
taken at various time intervals after a drug product is
administered.
The concentration of drug in each plasma sample is
plotted on rectangular-coordinate graph paper against
the corresponding time at which the plasma sample
was removed.
As the drug reaches the general (systemic)
circulation, plasma drug concentrations will rise up to
a maximum.
Usually, absorption of a drug is more rapid than
elimination. As the drug is being absorbed into the
systemic circulation, the drug is distributed to all the
tissues in the body and is also simultaneously being
eliminated.
Elimination of a drug can proceed by excretion,
biotransformation, or a combination of both.
MEC and MTC represent the minimum effective
concentration and minimum toxic concentration of
drug, respectively.
For some drugs, such as those acting on the
autonomic nervous system, it is useful to know the
concentration of drug that will just barely produce a
pharmacologic effect (ie, MEC).
Assuming the drug concentration in the plasma is in
equilibrium with the tissues, the MEC reflects
the minimum concentration of drug needed at the
receptors to produce the desired pharmacologic
effect.
Similarly, the MTC represents the drug
concentration needed to just barely produce a toxic
effect. The onset time corresponds to the time
required for the drug to reach the MEC.
The intensity of the pharmacologic effect is
proportional to the number of drug receptors
occupied, which is reflected in the observation that
higher plasma drug concentrations produce a greater
pharmacologic response, up to a maximum.
The duration of drug action is the difference
between the onset time and the time for the drug
to decline back to the MEC.
 In contrast, the pharmacokinetic can also describe
the plasma level–time curve in terms of such
pharmacokinetic terms as peak plasma level, time
for peak plasma level, and area under the
curve, or AUC.
 The time of peak plasma level is the time of
maximum drug concentration in the plasma and is
a rough marker of average rate of drug absorption.
 The peak plasma level or maximum drug
concentration is related to the dose, the rate
constant for absorption, and the elimination
constant of the drug. The AUC is related to the
amount of drug absorbed systemically.
Drug Concentrations in Tissues
Tissue biopsies are occasionally removed for
diagnostic purposes, such as the verification of a
malignancy.
Usually, only a small sample of tissue is removed,
making drug concentration measurement difficult.
Drug concentrations in tissue biopsies may not
reflect drug concentration in other tissues nor the
drug concentration in all parts of the tissue from
which the biopsy material was removed.
For example, if the tissue biopsy was for the
diagnosis of a tumor within the tissue, the blood flow
to the tumor cells may not be the same as the blood
flow to other cells in this tissue.
 In fact, for many tissues, blood flow to one part of
the tissues need not be the same as the blood flow
to another part of the same tissue.
 The measurement of the drug concentration in
tissue biopsy material may be used to ascertain if
the drug reached the tissues and reached the proper
concentration within the tissue.
Drug Concentrations in Urine and Feces
Measurement of drug in urine is an indirect method
to ascertain the bioavailability of a drug. The rate and
extent of drug excreted in the urine reflects the rate
and extent of systemic drug absorption.
Measurement of drug in feces may reflect drug that
has not been absorbed after an oral dose or may
reflect drug that has been expelled by biliary
secretion after systemic absorption.
Drug Concentrations in Saliva
Saliva drug concentrations have been reviewed for
many drugs for therapeutic drug monitoring.
Because only free drug diffuses into the saliva,
saliva drug levels tend to approximate free drug
rather than total plasma drug concentration.
The saliva/plasma drug concentration ratio is less
than 1 for many drugs. The saliva/plasma drug
concentration ratio is mostly influenced by the pKa
of the drug and the pH of the saliva.
Weak acid drugs and weak base drugs with pKa
significantly different than pH 7.4 (plasma pH)
generally have better correlation to plasma drug
levels.
 The saliva drug concentrations taken after
equilibrium with the plasma drug concentration
generally provide more stable indication of drug
levels in the body. The use of salivary drug
concentrations as a therapeutic indicator should be
used with caution and preferably as a secondary
indicator.
Forensic Drug Measurements
Forensic science is the application of science to
personal injury, murder, and other legal proceedings.
Drug measurements in tissues obtained at autopsy or
in other bodily fluids such as saliva, urine, and blood
may be useful if a suspect or victim has taken an
overdose of a legal medication, has been poisoned, or
has been using drugs of abuse such as opiates (eg,
heroin), cocaine, or marijuana.
The appearance of social drugs in blood, urine, and
saliva drug analysis shows short-term drug abuse.
These drugs may be eliminated rapidly, making it
more difficult to prove that the subject has been
using drugs of abuse.
The analysis for drugs of abuse in hair samples by
very sensitive assay methods, such as gas
chromatography coupled with mass spectrometry,
provides information regarding past drug exposure.
A study that the hair samples from subjects who
were known drug abusers contained cocaine and 6-
acetylmorphine, a metabolite of heroine
(diacetylmorphine).
Significance of Measuring Plasma Drug
Concentrations
The intensity of the pharmacologic or toxic effect of
a drug is often related to the concentration of the drug
at the receptor site, usually located in the tissue cells.
Because most of the tissue cells are richly perfused
with tissue fluids or plasma, measuring the plasma
drug level is a responsive method of monitoring the
course of therapy.
Clinically, individual variations in the
pharmacokinetics of drugs are quite common.
Monitoring the concentration of drugs in the blood or
plasma ascertains that the calculated dose actually
delivers the plasma level required for
therapeutic effect. With some drugs, receptor
expression and/or sensitivity in individuals varies, so
monitoring of plasma levels is needed to distinguish
the patient who is receiving too much of a drug from
the patient who is supersensitive to the drug.
Moreover, the patient's physiologic functions may
be affected by disease, nutrition, environment,
concurrent drug therapy, and other factors.
Pharmacokinetic models allow more accurate
interpretation of the relationship between plasma
drug levels and pharmacologic response.
In many cases, the pharmacodynamic response to
the drug may be more important to measure than just
the plasma drug concentration.
 For example, the electrophysiology of the heart,
including an electrocardiogram (ECG), is
important to assess in patients medicated with
cardiotonic drugs such as digoxin. For an
anticoagulant drug, such as dicumarol,
prothrombin clotting time may indicate whether
proper dosage was achieved. Most diabetic
patients taking insulin will monitor their own
blood or urine glucose levels.
Math Self-Exam
1. What are the units for concentration?
2. A drug solution has a concentration of 50 mg/mL.
What amount of drug is contained within 20.5 mL of
the solution? In 0.4 L? What volume of the solution
will contain 30 mg of drug?
3. If 20 mg of drug are added to a container of water
and result in a concentration of 0.55 mg/L, what
volume of water was in the container?
Exponents and Logarithms
In the expression

x is the exponent, b is the base, and N represents the

number when b is raised to the xth power, ie, b x. For
example,
where 3 is the exponent, 10 is the base, and 10 3 is the
third power of the base, 10. The numeric value, N in
Equation 2.1, is 1000. In this example, it can be
reversely stated that the log of N to the base 10 is 3.
Thus, taking the log of the number N has the effect of
"compressing" the number; some numbers are easier
to handle when "compressed" or transformed to base
10.
Transformation simplifies many mathematical
operations