MODULE 2: MEASUREMENTS OF DRUG CONCENTRATION

KNOWLEDGE OBJECTIVES:
1. Demonstrate knowledge of exponent, logarithms and graphs
2. Demonstrate knowledge of the basic systems of computing the drug concentrations

MODULE OUTLINE
A. Mathematical Fundamentals in Pharmacokinetics
1. Exponent and logarithms
2. Graphs – curve fitting and determination of slope

B. Drug Concentration and Its Significance

1. Measurements of drugs in biological fluids
2. Plasma Drug concentration-time relationship
3. Rates and Orders of reactions
4. Half-life

MATHEMATICAL FUNDAMENTALS IN PHARMACOKINETICS

A scientific calculator or computer software program with logarithmic and exponential functions
will make the calculations less tedious.

• Computer software is a tool that allows one to solve more complex pharmacokinetic
problems rapidly, but efficient use requires a thorough understanding of the subject.
Calculation accuracy is important!

• Calculators with exponential and logarithmic functions. Additional function: mean,

standard deviation, and linear regression analysis.

• Approximation a useful process for checking whether the answer to a given set of
calculations is probably correctly. To estimate a series of computations, round the
numbers and write the numbers using scientific notation.

Exponential and Logarithmic Functions

Exponent: is the power to which a number, symbol, or expression is to be raised.

For example, the 3 in x3.

Logarithm: The logarithm of a number is the exponent by which another fixed value, the base,
has to be raised to produce that number.
Log – logarithmic to base 10
ln (natural log) - logarithmic to base e

Graphs
Visualizing the relationship between variables.
Types of Graphs
- Cartesian or Rectangular Coordinate Graph Paper
- Semi-log graph paper

Trapezoidal Rule is a numerical method frequently used in pharmacokinetics to calculate the

area under the plasma drug concentration-versus time curve, called area under the curve (AUC).

Least square method a useful procedure for obtaining the line of best fit through a set of data
points by minimizing the deviation between the experimental and the theoretical line.

y = mx + b

Where:
y – the dependent variable
x – the independent variable
m – slope
b – y intercept

Curve Fitting is the process of constructing a curve, or mathematical function, that has the best
fit to a series of data points, possibly subject to constraints.
- It is a statistical technique use to drive coefficient values for equations that express the
value of one (dependent) variable as a function of another (independent variable).

Problems of Fitting Points to a Graph

➢ Consider the law of parsimony- “keep it simple”. That is, if a choice between two
hypotheses is available, choose the simpler relationship.

Interpolation – filling the gap between the observed data on a graph, is usually safe and assumes
that the trend between the observed data is consistent and predictable.

Extrapolation – predicting new data beyond the observed data, and assumes that the same trend
obtained between two data points will extend in either direction beyond the last observed data
points.

B. Drug Concentration and Its Significance

 • Drug concentrations are measured in biologic samples, such as milk, saliva, plasma, and
urine. Such measurement is generally validated so that accurate information is generated
for pharmacokinetic and clinical monitoring.

1. Measurements of drugs in biological fluids

General methods use measure drug concentrations by means of chromatographic which

separates drugs the drug from other related materials that may cause assay interference and
mass spectrometric allows detection of molecules or molecule fragments based on their mass-to-
charge ratio.
▪ Sampling of biologic specimens can be invasive include sampling blood, spinal fluid,
synovial fluid, tissue biopsy, or any biologic material that requires parenteral or surgical
intervention in the patient or noninvasive sampling of urine, saliva, feces, expired air, or
any biologic material that can be obtained without parenteral or surgical intervention.

▪ 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.

▪ Drugs in the plasma are often bound to plasma proteins are filtered from the plasma before
drug concentrations are measured. This is the unbound drug.

▪ Alternatively, drug concentration may be measured from unfiltered plasma; this is the total
plasma drug concentration.

2. Plasma Drug concentration-time relationship

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 circulation, plasma drug concentrations will rise up to a
maximum if the drug was given by an extravascular route.
Usually, absorption of a drug is more rapid than elimination. Elimination of a drug can proceed by
excretion, biotransformation, or a combination of both.
Drug product performance parameters:

MEC a minimum effective concentration of drug needed to produce the desired pharmacologic
effect.
MTC minimum toxic concentration represents the drug concentration needed to produce toxic
effect.
Onset time the time required to reach the MEC.
Duration of Drug Action – the difference between the onset time and the time for the drug to
decline back to the MEC.
Therapeutic Window – concentrations between the MEC and the MTC (therapeutic index).

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 in urine is an indirect method.
The extent of drug excreted in the urine reflects the rate and extent of systemic drug
absorption.
While drug in feces may reflect drug that not been absorbed after an oral dose or may
reflect drug that has been expelled by biliary secretion after systemic absorption.

Only free drug diffuses into the saliva, saliva drug levels tend to approximate free drug rather than
total plasma drug concentration.
▪ Mostly influenced by the pka of the drug and the pH of the saliva.
▪ The used of salivary drug concentrations as a therapeutic indicator should be used with
caution and preferably as a second indicator.

Forensic drug measurement is the application of science to personal injury, murder, and other
legal proceedings. Drug measurement 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.

Significance of Monitoring Plasma Drug Concentrations

1. 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.

2. Measuring the plasma drug level is a responsive method of monitoring the course of
therapy.

3. Ascertains that the calculated dose actually delivers the plasma level required for
therapeutic effect.
4. 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.

5. The patient’s physiologic functions may be affected by disease, nutrition, environment,

concurrent drug therapy, and other factors.

6. Allows for the adjustment of the drug dosage in order to individualize and optimize
therapeutic drug regimens.

7. Provide a guide to the progress of the disease state and enable the investigator to modify
the drug dosage accordingly.

8. Therapeutic decisions should not be based solely on plasma drug concentrations.

9. Pharmacodynamic response to the drug is more important to measure than just the
plasma concentration.

RATES AND ORDERS OF REACTIONS

The RATE of a chemical reaction velocity with which it occurs

The ORDER of a reaction the way in which the concentration of a drug or reactant in a chemical
reaction affects the rate. Refers to the way in which the concentration of a drug or reactants
influences the rate of a chemical reaction or process

CLASSES:
A. Zero-order reactions the drug concentration changes with respect to time at a constant rate

C = -k0t + C0

Where:
C = drug concentration at any time
K0 = zero-order rate constant (units of concentration per time) is the slope of the line
C0 = is the y intercept drug concentration, when time (t) equals zero
Negative sign = indicates that the slope is decreasing

B. First-order reactions
The Cp vs Time profile during the elimination phase is linear
Example: 1.2 mg are eliminated every hour, independently of the drug concentration in the body.

▪ The drug concentration changes with respect to time equal to the product of the rate
constant and the concentration of drug remaining.
▪ A first order process is where the amount of drug eliminated may change with the amount
of drug in the body, but the fraction of a drug in the body eliminated over a given time
remains constant

Fraction or percent of drug being removed is the same with a high or a low drug concentration

Formula:
C = C0e-kt
ln C = -kt + ln C0
log C = -kt + log C0
2.3
Where:
C = drug concentration at any time
k = first-order rate constant (units of reciprocal time, or time-1)
-k/2.3 = is the slope of the line
C0 = is the y intercept = drug concentration, when time (t) equals zero

First order kinetics means that the rate of change of drug concentration by any process is
directly proportional to the drug concentration remaining to undertake that process.
In first-order elimination the amount of drug eliminated in a set of time is directly
proportional to the amount of drug in the body

Zero Order elimination is rare mostly occurring when the elimination system is saturated. An
example is the elimination of Ethanol.

Significance of rate constants

Characterize the change of drug concentration in a particular reference region.

Give the speed at which a drug:

Enters the compartment (absorption rate constant, ka)
Distributes between a central and peripheral compartment (distribution rate constant)
Is eliminated from the systemic circulation (elimination rate constant, k)

HALF-LIFE (t½) Units: time

Expresses the period of time required for the concentration of a drug to decrease by one half
Also define as the time required to decrease the initial dose of drug by 50% (one half of original
value).

Significance of Half-life

Determine the dosing interval necessary to obtain the desired CP of the drug
Generally, the dosing interval is the same as t½

Predict how long it will take a drug to reach steady-state levels

Predict the accumulation of a drug in the body for a specific dosing interval
 During multiple dosing or continuous IV infusion it takes approximately 4-5 half-
lives to reach steady-state levels

Predict how long it will take a drug concentration to decrease to a lower concentration
All drugs are decreased by 96% after 4 half-lives