Physical Pharmacy

Biopharmaceutics

The study of the physical and chemical properties of

drugs and their proper dosage as they relate to
LADME

Study of the effects of the physicochemical

properties of the drug and the drug product, in vitro,
on the availability of the drug, in vivo, to produce a
desired therapeutic effect
in vitro vs in vivo

in vitro refers to the technique of performing a

given procedure in a controlled environment outside
of a living organism

in vivo ("within the living); are those in which

the effects of various biological entities are tested on
whole, living organisms usually animals including
humans, and plants

Physicochemical properties of the drug

The drugs biological fate in the body after it is taken

by the patient

Resulting pharmacological action of the drug

How do drugs exert their effects?

Targets
o Biological targets
o Molecules where drugs act on, thus
establishing their pharmacological effects
o Normally a macromolecule
Targets are normally proteins:

Enzymes

Hormone and neurotransmitter receptors --a

particularly prominent class are the G proteincoupled receptors (GPCRs).

Ion channels

Membrane transporters

Cytoskeletal proteins

Non-protein targets include

DNA: alkylating anti-tumor drugs

RNA: anti-ribosomal antibiotics

Lipid membranes: antibiotics (amphotericin B,

polymyxin)
Physicochemical factors related to the fate of the drug in the
body

Solid state properties

Ionization

Solubility and dissolution

Partition coefficient

Mass transport and membrane passage

Drug transporters

Complexation and protein binding

BINDING FORCES

Will dictate the state of matter

Intermolecular Forces vs Intramolecular Forces

Intermolecular forces

Intermolecular binding forces

o Attraction vs repulsion
o Attractive forces: cohesive + adhesive
- Inversely proportional to the distance
separating the molecules
- As the forces of attraction between the
molecules increase, the potential energy
becomes increasingly negative ( LennardJones potential theory)

Intermolecular attractive forces:

o dipole-dipole (Keesom)
o Dipole-induced dipole (Debye)
o Induced dipole-induced dipole (London)
o Ion-dipole

Hydrogen bonds
o Inter- or intra- molecular bonds
o Attraction of a hydrogen atom for a
strongly electronegative atom
o Important for establishing the secondary
structure of proteins
o Important in the structure of DNA
o Involves interaction between a molecule
containing H and a highly electronegative
atom (F, O, N)
o Responsible for the many unusual property
of water

o
o

High dielectric constant (property of a

solvent to separate the charges of the
solute)
Abnormally low vapor pressure

Hydrophobic interactions
o Between nonpolar atoms and molecules in
water
o Nonpolar = water-fearing

States of Matter: Gases

Molecules that have higher kinetic energy that

produce rapid motion

Held together by weak intermolecular forces

No regular shape

Compressible

Invisible (mostly)

Charles's law states that volume and temperature are

directly proportional to each other as long as
pressure is held constant.

Boyle's law asserts that pressure and volume are

inversely proportional to each other at fixed
temperature.

Gay-Lussac's law introduces a direct proportionality

between temperature and pressure as long as it is at a
constant volume.
States of Matter: Liquids

A liquid occupies a definite volume and takes the

shape of the container required to hold it

Denser than gas

Possess less kinetic energy than gases

Flow readily, and the flow is influenced by friction

Vapor pressure

Surface tension
States of Matter: Solids

Fixed shape

Incompressible

Main types: crystalline, amorphous and polymeric

EXP1: SPECIFIC GRAVITY DETERMINATION OF
LIQUIDS
PYCNOMETER or Specific Gravity Bottle
A pycnometer is a special glass bottle used to determine
specific gravity. Pycnometers are generally available for
laboratory use in volumes ranging from 1 mL to 50
mL.Pycnometers have fitted glass stoppers with a capillary
opening to allow trapped air and excess fluid to escape. Some
pycnometers have thermometers affixed in order to relate the
specific gravity, as determined, with temperature.

EXP1: SPECIFIC GRAVITY DETERMINATION OF

LIQUIDS
PYCNOMETER or Specific Gravity Bottle

A pycnometer is a special glass bottle used to

determine specific gravity. Pycnometers are
generally available for laboratory use in volumes
ranging from 1 mL to 50 mL.Pycnometers have
fitted glass stoppers with a capillary opening to
allow trapped air and excess fluid to escape. Some
pycnometers have thermometers affixed in order to
relate the specific gravity, as determined, with
temperature.

In using a pycnometer, it is first weighed empty and

then weighed again when filled to capacity with
water. The weight of the water is calculated by
difference. Since 1 g of water equals 1 mL, the exact
volume of the pycnometer becomes known. Then,
when any other liquid subsequently is placed in the
pycnometer, it is of equal volume to the water, and
its specific gravity may be determined.
HYDROMETER

A hydrometer is an instrument used to measure the

density of a liquid as compared to that of water.
Hydrometers usually consist of a calibrated glass
tube ending in a weighted glass sphere that makes
the tube stand upright when placed in a liquid.

The greater the density, the tighter or closer the

molecules are packed inside the substance.

Therefore, the greater the density / specific gravity

of a liquid the higher a hydrometer will be buoyed
by it.

Fill your hydrometer jar about with the liquid you

wish to test. Insert the hydrometer slowly. Do not
drop it in! Now give it a spin with your thumb and
index finger, this will dislodge any bubbles that may
have formed. Once the hydrometer comes to a rest,
observe the plane of the liquid surface. Your eye
must be horizontal to this plane. The point at which
this line cuts the hydrometer scale is your reading.
TYPES OF HYDROMETER:

Specific Gravity hydrometers can be used for almost

any liquid. Specific Gravity is a dimensionless unit
defined as the ratio of density of the material to the
density of water. If the density of the substance of
interest and the reference substance (water) are
known in the same units (e.g., both in g/cm3 or
lb/ft3), then the specific gravity of the substance is
equal to its density divided by that of the reference
substance (water =1 g/cm3)

Baume hydrometers are calibrated to measure

specific gravity on evenly spaced scales; one scale is

for liquids heavier than water, and the other is for

liquids lighter than water.
These two scales, one for liquids lighter than water
and one for liquids heavier than water, were
developed by the French chemist Antoine Baum in
1768. It is widely used in industrial
chemistry, pharmacology, sugar refining and other
industries.
Brix (BX) hydrometer is for determining the
percentage of weight by sucrose. One degree Brix is
1 gram of sucrose in 100 grams of solution and
represents the strength of the solution as percentage
by weight (% w/w) (strictly speaking, by mass). If
the solution contains dissolved solids other than pure
sucrose, then the Bx only approximates the
dissolved solid content. The Bx is traditionally used
in the wine, sugar, fruit juice, and honey industries.
MOHR/WESTPHAL BALANCE
o The principle of the balance is based on
the known buoyancy of a reference glass
body.
o 1 Foot with adjustment screws
o 2 Metal frame with adjustable height
o 3 Balance beam with notches
o 4 Plummet (glass cylinder with thin
Platinum wire)

EXPERIMENT 2: MELTING POINT DETERMINATION

The temperature at which a solid melts and becomes

a liquid is the melting point. Since this requires that
the intermolecular forces that hold the solid together
have to be overcome, the temperature at which
melting occurs will depend on the structure of the
molecule involved - an example of the relationship
between structure and properties. Hence, different
compounds tend to have different melting points.
The Melting Point as a Criterion of Purity

A dilute solution of a liquid begins to freeze at a

temperature somewhat lower than the freezing point
of the pure liquid. The presence of an impurity
causes a reduction of the freezing point of the
sample. As the pure solvent crystallizes from
solution, the concentration of the impurity must
increase and the freezing point of the solution must
fall. Thus, a sharp melting point (actually, a melting
range of less than about 1C) is often taken as
evidence that the sample is fairly pure, and a wide
melting range is evidence that it is not pure.
The Melting Point as a Means of Identification and
Characterization

If two samples have different melting points, their

molecules must differ either in structure or in
configuration. They must be either structural isomers
or diastereomers. If the melting points of two
samples are the same, the structures of their
molecules must be the same, although they might
have enantiomeric configurations. These statements
apply only to pure substances, and do not take into
account the fact that some substances can exist in
different crystalline forms that have different melting
points.

Polarity

As with boiling points, compounds with polar

functional groups generally have higher melting
points than compounds with nonpolar functional
groups. In contrast to the case with boiling points,
highly branched or cyclic molecules (relatively
symmetrical molecules) tend to have higher melting
points than their straight-chain isomers. The
combined effects of branching or the presence of
rings, then, are to reduce the range of temperature
over which the liquid can exist at a vapor pressure of
less than 760 Torr. In extreme cases, a liquid range
does not exist at a vapor pressure of less than 760
Torr; at atmospheric pressure, the substance will
sublime without melting.
EXP 3: REFRACTIVE INDEX DETERMINATION

A refractometer measures the extent to which light is

bent (i.e. refracted) when it moves from air into a
sample and is typically used to determine the index
of refraction (aka refractive index or n) of a liquid
sample.

The italicized n denotes refractive index,

the superscript indicates the temperature in degrees
Celsius, and the subscript denotes
the wavelength of light (in this case the D indicates
the sodium D line at 589 nm).
The refractive index is commonly determined as
part of the characterization of liquid samples,
in much the same way that melting points are
routinely obtained to characterize solid compounds.
It is also commonly used to:

Help identify or confirm the identity of a sample by

comparing its refractive index to known values.

Assess the purity of a sample by comparing its

refractive index to the value for the pure substance.

Determine the concentration of a solute in a solution

by comparing the solution's refractive index to a
standard curve.

Acids and bases can be divided into two broad

categories, strong and weak.
Strong acid lose their acidic protons virtually 100 %
and strong bases accept protons virtually 100 %.
Weak acids and bases lose and gain protons
respectively less than 10 %.
All the acids and bases described above
would be considered to be weak.
Note:

Fig. 1.
Refraction
The speed of lightof
in alight
substance is slower than in a

vacuum since the light is being absorbed and

reemitted by the atoms in the sample. Since the
density of a liquid usually decreases with
temperature, it is not surprising that the speed of
light in a liquid will normally increase as the
temperature increases. Thus, the index of refraction
normally decreases as the temperature increases for a
liquid.
For many organic liquids the index of refraction
decreases by approximately 0.0005 for every 1 C
increase in temperature. However for water the
variation is only about -0.0001/C.

EXPERIMENT 4: BUFFER SOLUTIONS

pH is a measure of the concentration of H+ [H3O+]

ions in a solution. Only the concentration of H+ and
OH- molecules determine the pH. When the
concentration of H+ and OH- ions are equal, the
solution is said to be neutral. If there are more
H+ than OH- molecules the solution is acidic, and if
there are more OH- than H+ molecules, the solution
is basic.
A buffer is a solution that resists change in pH upon
addition of acid, addition of base, and upon dilution.
Buffer solutions contain significant quantities of
both partners of a Bronsted-Lowry conjugate acid-base pair.

How does a mixture of a weak acid and its conjugate base help
buffer a solution against pH changes?
So, the weak acid and weak base remain in the
solution with high concentrations since they only rarely react
with the water. However, they are very likely to react with any
added strong base or strong acid.

If a strong base is added to a buffer, the weak acid will give up

its H+ in order to transform the base (OH-) into water (H2O)
and the conjugate base: HA + OH- A- + H2O. Since the
added OH- is consumed by this reaction, the pH will change
only slightly.

According to Bronsted and Lowry an acid is a proton (an H+ )

donor. A base is definedas a substance that can accept a proton.
When an acid gives up a proton, a species that can accept a
proton, a base, is formed from the acid. That base is called the
conjugate base of the acid.
HA -----------------------> H+ + A Acid
Conjugate Base
Although we frequently represent the proton in water
(aqueous) solution as H+, actually the proton combines with
water to form ions like H3O + , H5O2+ , and H7O3+. The
main form is the hydronium ion, H3O+.

If a strong acid is added to a buffer, the weak base will react

with the H+ from the strong acid to form the weak acid HA:
H+ + A- HA. The H+ gets absorbed by the A- instead of
reacting with water to form H3O+ (H+), so the pH changes only
slightly.

EXPERIMENT 4: BUFFER SOLUTIONS

When working with a buffer one must be concerned with two
major questions:
1. Over what pH range will the buffer work and
2. What is the capacity of this buffer to resist pH
change?

The initial pH of a buffer depends upon two factors:

o the strength of the weal acid or weak base
and
o the ratio of weak acid to its conjugate base.

Considering the first fact, the stronger the weak acid

the more acidic will be the pH of the buffer;

The weaker the weak acid the more basic the initial
pH.
After the range is determined by the strength of the weak acid
component, the actual initial pH is determined by the ratio of
the weak acid to the conjugate base.

The pH of the buffer will be a bit more basic if more

of the conjugate base is present than the weak acid,
and it will be a bit more acidic if more of the weak
acid is present than the conjugate base.
The buffer capacity depends upon two factors:
1. Concentration of the buffer

In general the more concentrated the buffer, the more

ingredients are available to attack added H+ and OH
- ions.
2. What is being added to the buffer and how much of
each component, acid and conjugate base, is
available to react.

Suppose that a buffer had 100 times as much acetic

acid as it had acetate.

This buffer could resist a challenge by base because

there would be plenty of acetic acid to react with the
base.

It would not, however, be able to resist an attack by

acid, because there would be relatively little acetate
to react with the acid.