Distribution Phenomena

Oil Oil

Water Water

Co
K o/w =
Cw
Distribution and Partitioning: Quantitative Description
• When two immiscible liquids are added to each other they
form two separate and distinct phases.
• Systems of water and ether or amyl alcohol, octanol or
peanut oil are examples of such liquid systems.
• The movement of a solute from one phase to another is
called partitioning.
• If the substance is added in an amount insufficient to saturate
the solutions, it will distributed between the two layers in a Oil
definite concentration ratio.
• If an excess solid or liquid is added to such a system, it will
distribute between the two phases so that each phase K

become saturated (the ratio of its concentration in each

phase is equal to the ratio of its solubility limit)
• If Co and Cw are the equilibrium concentration of the
substance in oil phase and water phase respectively, the Water
equilibrium expression is: Co So lubilityo
K o/w = ≈
Cw So lubilityw
• The above equation is known as the distribution law.
• The equilibrium constant Ko/w is known as the distribution or
partitioning coefficient between oil and water (respectively. It
is different than Kw/o!!)
 The partition coefficient is measures of how
hydrophilic ("water-loving") or hydrophobic ("water-
fearing") a chemical substance is

Hydrophobic Drug Hydrophilic Drug

High Ko/w Low Ko/w

Oil Oil Oil

K K

Water Water Water

Pharmaceutical Relevance of Distribution
and Partitioning: Intestinal permeability
and drug absorption

Low Ko/w Optimized Ko/w High Ko/w

 the molecules nearest to the skin surface enter the cellular lipids.
tissue these must be replaced by other molecules In contrast, the intercellular route requires the
Pharmaceutical Relevance of Distribution and
diffusing within the formulation towards the skin molecule to partition into the lipid bilayers between
Partitioning: Drug absorption and distribution of drugs
throughout the body
Partitioning of drug FORMULATION
into skin layers Drug dis s olution
Depend onKo/w Drug diffus ion to s kin s urfa ce
INTRACELLULAR ROUTE INTERCELLULAR ROUTE S HUNT ROUTES
P a rtition into P a rtition into P a rtitioning into
come ocyte lipids s we a t or s e bum
Diffus ion
Pa rtition Diffus ion
S tra tum Diffus ion Diffus ion Diffus ion Drug ma y bind
corne um P a rtition Diffus ion

Via ble P a rtition P a rtition Diffus ion P a rtitioning

Drug ma y be
e pide rmis into via ble into via ble through via ble into via ble
me ta bolize d
e pide rmis e pide rmis e pide rmis e pide rmis

Ca pilla ry P a rtitioning
upta ke into de rmis

De rmis Eccrine s we a t duct

Ha ir follicle

Ca pilla ry
 Distribution and Partitioning: Methods of determination
P h a rm a c e u tic a l p re fo rm u la tio n C H AP TE R 2 3
For Non-electrolyte Drug
Determination of log P Conce ntra tion
be fore s ha king
Conce ntra tion
a fte r s ha king
• TheLogmost
P valuescommonly usedexperimentally
can be determined method for or
thecandetermination
be calculated rom the ofchemical
the partitioning
structure o the
0 C wa te r, initia l–C wa te r, fina l
drug candidate using group additivity unctions. For
coefficient of a there
the latter approach drug are is the Shake-
numerous computer
Flask
modelsmethod.
and simulation methods available; selection
will reduce to personal choice and amiliarity and so
• A these
drug modelswhose
will not be K o/w is here.
considered to Rather,
be C wa te r, initia l C wa te r, fina l
this text will ocus on experimental determination.
determined is traditionally added
It is clear, however, that there is much value to be
initially
gained by tocomparing
the water (or and
calculated sometime to
experimentally
thedetermined
oil) log P values. The value o the calculation
phase in a to a separating
approach is greatest when selecting a lead candidate
funnel
rom a followed
compound libraryby addition of simply
when it would the oilbe
neither possible, nor practicable to measure the par-
phase (n-octanol) and vigorous
titioning behaviour o the many thousands o com-
shaking (usually 30 minutes).
pounds available.
Fig . 23.9 • The shake ask method for determination of
partition coef cient.
• theShaconcentration
ke - a s k me thod of drug in each
phase
Assuming is thatdetermined and K
a UV assay is available, o/w
the is
shake- or partition coe f cient needs to be corrected to
calculated.
ask method is a quick, simple and near universally account or the di erent volumes. For example,
assuming a 1 : 9 n-octanol:water ratio, Equation
applicable way o determining the partition coe f -
cient (see also Fig. 21.2). Prior to measurement, the 23.18 becomes:
 Distribution and Partitioning: Methods of determination
For Weak-electrolyte Drug

• Shake-Flask method is used. However,

ionization may occur in the aqueous
phase. Remember that partition is
limited to the unionized form and that
the analysis of the aqueous layer will
count both ionized and un-nionized
forms. For this reason, we need to
suppress (prevent) the ionization in
the aqueous layer by selecting a
proper pH. To ensure complete
UNIONIZATION:
we use pH=pKa-2 for weak acid
and pH=pKa+2 for weak base
 Weak acid: HA ……………..>A- + H+
Weak base: B + H+……………..> BH+

BH+ A- Henderson-Hasselbalch Equation

Cunionized
Generally : pK a − pH = log
Cionized
[HA]w
acids : pK a − pH = log −
[A ]w
HA B beases : pK a − pH = log
[B]w
[BH + ]w
 For weak electrolyte with adjusted
For Non-electrolyte pH to suppress the ionization
solute (pH=pKa-2) for weak acids and
(pH=pKa+2) for weak bases
Co [A]o Co [HA]o
K o/w = = K o/w = =
Cw [A]w Cw [HA]w

Oil Oil

[A]o [HA]o

K K

[A]w [HA]w
Ionization is
suppressed by Water
Water
low pH in the
case of weak
acid here
 If we can not stop dissociation:
We need to learn more on the effect of ionic
dissociation on partitioning
• Many pharmaceutical preparations and
biological compartments are buffered at
physiological pH values (typically=7.4) so for
weak acids or bases, ionization is expected.
e.g.: 50% of phenobarbital (pKa=7.4) will be
ionized in blood!!
• Benzoic acid (pKa=4.2) exist in two forms: an
ionized (dissociated) form and an associated
(unionized) form in water (upon addition of
benzoic acid, pH will be slightly acidic around 5)
• In the flask to the right, only the unionized
form of the benzoic acid (called the “common”
specie) will partition to the oil phase in a
water/peanut oil system
 “True” versus “Apparent” Distribution Coefficient

If No dissociation!!
Co [HA]o
K o/w = =
Cw [HA]w
Ko/w is the true distribution coefficient Oil
[HA]o
In case of Dissociation!! O
Benzoic acid concentration obtained by the
analysis of the aqueous phase is total OH
benzoic acid concentration of both ionized Ko/w
and unionized forms
O O
C [HA]o Ka
+ H+
K 'o/w = o =
Cw [ HA]w + [ A −]w [HA]w
OH
[A-]w
O

[HA]o [HA]w
K 'o/w = × Water
[ HA]w [ HA]w + [ A −]w
K 'o/w = K(true) * F(unionized )
K'o/w is the apparent or observed distribution coefficient
How to determine true distribution coefficient?

The true distribution coefficient, Ko/w , can be obtained:

1. at a low pH (i.e. 2) at which the acid would exist completely in the un-
ionized form.
Co [HA]o
K o/w = =
Cw [HA]w
2. Or, alternatively, Ko/w can be obtained from the linear equation:

ka + !" H 3O + #$ ka K o/w +1
= + [H 3O + ]
Cw C C

Y = a + bX
§Assumption: equal volumes of oil and water
§Cw: Drug concentration In aqueous phase at equilibrium ( [AH]w+[A-]w )
§C: initial concentration of drug in water phase (or oil phase) before distribution.
§In this case, the oil phase need not to be analyzed: only the hydrogen ion concentration (pH)
and Cw in the aqueous phase.
 Cw [ HA]w + [ A −]w Cw [ HA]w + [ A −]w Cw [ HA]w + [ A −]w Cw [ HA]w + [ A −]w

Oil Oil Oil Oil

[HA]o [HA]o [HA]o [HA]o

K K K K
Ka Ka Ka Ka
[HA]w [A-]w+ [H+] [HA]w [A-]w+ [H+] [HA]w [A-]w+ [H+] [HA]w [A-]w+ [H+]

Water Water Water Water

pH=1.2 pH=4.5 pH=5.5 pH=7.5

Cw=5mg/mL Cw=6.3mg/mL Cw=7.1mg/mL Cw=8.2mg/mL

ka + [ H 3O +] ka K o/w +1 ka + [H 3O +]
= + [H 3O+] Cw
Cw C C
! !
Cw! pH! [ H 3O +] ka + [ H 3O +]
! Cw
b= (K+1)/C

Intercept = ka/C

[ H 3O +]
 How to determine true distribution coefficient?
Example 9-6 page 194 (Martin’s Physical Pharmacy 6th ed.):
ka + !" H 3O + #$
Find true partition coefficient
Cw (Ko/w)

ka + !" H 3O + #$ ka K o/w +1
= + [H 3O + ]
Cw C C
ka
K +1
slope = b = o/w = 4.16 a = → C = 1.52
C C
K +1
b = o/w → K o/w = 5.31
C

[H 3O+ ]
ka
int ercept = a = = 4.22 x 10 -5
C

Ka= 6.4 x 10-5

Summary
Weak-electrolyte solute
For Non-electrolyte For weak electrolyte with adjusted pH to
solute suppress the ionization (pH=pKa-2) for Co [HA]o
weak acids and (pH=pKa+2) for weak
K o/wFor
= weak= electrolyte
bases withCionization
w [HA]w
Co [A]o Co [HA]o
K o/w = = C [HA]o K 'o/w = =
Cw [A]w K o/w = o =
Cw [HA]w Cw [ HA ]w + [ A −]w

Oil Oil Oil

[A]o [HA]o [HA]o

K K K
[A]w Ka
[HA]w [HA]w [A-]w+ [H+]

Water Water Water

 Pharmaceutical Relevance of Partitioning:
preservation of oil-water systems (emulsions,
creams,..etc)
Preservative
molecules
Partition into
the oil phase

Oil
droplets Preservative molecules
Preservative molecules are lost in the oil phase
(red dots) are initially leaving the aqueous phase
added with low levels of
to the water phase preservative and thus
spoilage may occure!!
Preservative or bacteriostatic
action of benzoic acid is due
almost entirely to the
undissociated form and not to the
ionic form. Therefore, the efficacy
of a preservative depends on the
concentration of the undissociated
acid in the aqueous phase

O O O
OH OH O

+H

[HA]o [HA]w [A-]w

Oil Phase Water Phase

 Calculating final concentration of preservative [HA]w stay in water
phase after partitioning as function of (K0/w; [H3O+]; Ka; q)
m = mo + mwm: original amount of preservative in the water phase
CVw = CoVo + CwVw (divide by Vw)
Vo
C = Co + Cw
Vw
C = Coq + Cw (use Co = [HA]o = Ko/w[HA]w)
C = K o/w q[HA]w + Cw Cw (use Cw = [HA]w + [A-]w)
Partitioning into C = K o/w q[HA]w +[HA]w +[A − ]w (use [A-]w= Ka [HA]w/[H3O+])
the oil phase
ka[HA]w (extract the common
C = K o/w q[HA]w +[HA]w +
"# H 3O + $% Ka [HA]w/[H3O+])
ka
C = [HA]w {K o/w q +1+ } (reorder)
"# H 3O $%
+

C This will allow us to calculate

[HA]w = the concentration of the
ka unionized form of BA “the
K o/w q +1+
"# H 3O + $% active” preservative
Calculating final concentration of preservatives [HA]w stay in water
phase after partitioning as function of (K0/w; [H3O+]; Ka; q)
Example:
•How much benzoic acid, Ka = 6.4 X10-5, will remain
undissociated in the aqueous phase of a 50% oil-water emulsion
if the initial concentration of benzoic acid in the aqueous phase
is 0.5%? The aqueous phase is buffered at pH 5 and the o/w
partition coefficient = 5.33. Assume that benzoic acid remains as
a monomer in the oil phase.
Given
C
[HA]w =
C= 0.5% (0.5g/100mL) ka
K0/w= 5.33 K o/w q +1+
!" H 3O + #$
[H3O ]= 10
+ -5

Ka=6.4X10-5
0.5g /100mL
[HA]w =
6.4X10 −5
5.33+1+
10 −5
[HA]w = 0.3928mg /100mL = 0.039%
 Extraction
• Extraction of a compound from a
solvent is an operation commonly
employed in analytical chemistry,
organic chemistry and
phytochemistry.
• Extraction depends on the partition
and distribution properties of the
extracted solute.
Co So lubilityo
K o/w = ≈
Cw So lubilityw
• K should be >>1 or <<1 for
separation. If K = 1, we don’t get a
good separation.
 O

Water-insoluble
acid,base,neutral OH

In the
organics organic
Acidified
Initially in the oil layer
aqueous
phase
layer

O
In the
OH aqueous
N H layer
N

How you can In the

separate organic
the three layer
compounds?

Controlling the Basified

pH; Selective aqueous O In the
extraction!! layer O
aqueous
layer
 Extraction Efficiency:
Calculating: [Solute remains in the first solvent, mw]
[solute appears in the extracting solvent, mo]
m = mo + mw
mw
Cw =
Vw
mo m − mw Oil Oil
Co = = mo m − mw
V0 V0 m0 = m - mw Co = =
Vo V0 V0
Cw " mw % Vo
K w/o = =$ '×
Co # m − mw & Vw m mw
Vw Vw mw
Cw =
K w/oVw Vw
mw = m Water Water
K w/oVw +V 0
The process can be repeated for n times
and the weight remaining after n
repetitions is (mw ’ n): Oil Oil Oil Oil

n
! K w/oVw $
(mw, n) = m # &
" K w/oVw +V 0 % Water n=1 Water n=2 Water n=3 Water
 Extraction Efficiency:
Calculating: [Solute remains in the first solvent, mw]
[solute appears in the extracting solvent, mo]

K1/2V 1
mremain1 = m
K1/2V 1 +V 2
Oil 2
Phase Oil 2
Phase
K w/oVw
mw = m [initalliy..in..water] m2 = m – m1
K w/oVw +Vo V2
K o/wVo m m1
mo = m [initalliy..in..oil]
K o/wVo +Vw V1 V1
Phase
Water1 Phase
Water1

1
K o/w =
K w/o
 Extraction: Example A.1
Calculating amount extracted and amount remained!!
The distribution coefficient for iodine between water and CCl4 at
25oC is Kw/o = 0.012
A) How many grams of iodine are extracted from a solution in water
containing 0.1 gm in 50 ml by one extraction with 10 ml of CCl4?

K w/o = 0.012 mw = m K w/oVw

K w/oVw +V 0 Oil Oil
m = 0.1g
0.012 * 50 m0 = m – mw=?
Vw = 50mL mw = 0.1g Vo=10mL
0.012 * 50 +10
Vo = 10mL
mw = 0.0057g m=0.1 g mw=?
mw = ?? Vw=50 Vw=50mL
mo = m − mw
mo = ?? Water Water
mo = 0.1− 0.0057 = 0.0943g
The Initial amount of iodine in water was 0.1 gm
The amount of iodine remained in water is 0.0057 gm
The amount of iodine extracted to CCl4 is 0.0943 gm
 Extraction: Example A.2
Calculating amount extracted and amount remained!!
The distribution coefficient for iodine between water and CCl4 at
25oC is Kw/o = 0.012
A) How many grams of iodine are extracted from a solution in water
containing 0.1 gm in 50 ml by one extraction with 5.0 ml of CCl4?

K w/o = 0.012 mw = m K w/oVw

K w/oVw +V 0 Oil Oil
m = 0.1g
0.012 * 50 m0 = m – mw=?
Vw = 50mL mw = 0.1g Vo=5.0 mL
0.012 * 50 + 5
Vo = 5.0mL
mw = 0.0107g m=0.1 g mw=?
mw = ?? Vw=50 Vw=50mL
mo = m − mw
mo = ?? Water Water
mo = 0.1− 0.0107 = 0.0893g
The Initial amount of iodine in water was 0.1 gm
The amount of iodine remained in water is 0.0107 gm
The amount of iodine extracted to CCl4 is 0.0893 gm
 Extraction: Example A.3
Calculating amount extracted and amount remained!!
The distribution coefficient for iodine between water and CCl4 at
25oC is Kw/o = 0.012
A) How many grams of iodine are extracted from a solution in water
containing 0.1 gm in 50 ml by two 5 ml portions of CCl4?
n
K w/o = 0.012 ! K w/oVw $
mw = m # &
m = 0.1g " K w/oVw +V 0 %
Vw = 50mL ! (0.012 * 50) $
2 Oil Oil Oil
m0 = m - mw
Vo = 5.0mL mw = 0.1g #" (0.012 * 50) + 5 &% Vo=5mL Vo=5mL Vo=5mL
n=2 Vw=50mL Vw=50mL Vw=50mL
mw = 0.0011g m mw
mw = ?? Water n=1 Water n=2 Water
mo = m − mw
mo = ?? mo = 0.1− 0.0011 = 0.0989g
The Initial amount of iodine in water was 0.1 gm
The amount of iodine remained in water is 0.0011 gm
The amount of iodine extracted to CCl4 is 0.0989 gm
 Extraction
Calculating amount extracted and amount remained!!
• The most efficient extraction results when n is large and V2 is small
(i.e. large number of extractions using small volumes of extracting
solvent)

Vo Amount remained in the Amount extracted to

(mL) n water phase (mw) the oil phase (mo)
Total volume used is

10 1 0.00566 0.094339623
constant=10 mL

5 2 0.001148 0.098852041

2 5 6.54E-05 0.099934553

1 10 5.5E-06 0.099994501
0.5 20 5.43E-07 0.099999457
 Extraction: Example A.3
Calculating amount extracted and amount remained!!
Example (Homework):
If 0.15 g of succinic acid in 100 mL ether is shaken with:
a)One (100) mL lot of water.
b)10 successive (10) mL portions of water.
Calculate the percentage recovery in each case. Ko/w = 0.125 at 25°C.

Answer:
a)89%
b)99.7%

Example (Homework):
• If 0.15g of succinic acid in 100ml of ether is shaken with a 10-ml portion of water, how much
succinic acid is left in the ether layer? The distribution coefficient K= (concentration in
ether)/(concentration in water) = 0.125 at 25°C. How much succinic acid is left in ether when
the phase is extracted with an additional 10ml of water?

Answer: 0.083g after the first extraction; 0.046g after the

second extraction