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BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A.

Samanta 1

CHAPTER – 1
Preformulation Studies
Syllabus:
Preformulation Studies: pKa and solu8bility, partition coefficient, crystal morphology, polymorphism, powder flow,
structure characteristics, dissolution, compatibility studies, protocol for pre-formulation..

Questions
Q1. Define polymorphism. [2]
Q2. Relevance of crystal morphology in solutions. [2]
Q3. Importance of powder flow in solid dosage form. [2]
Q4. Importance and application of dissolution.
Q5. Explain the importance of pKa and partition coefficient in the preformulation studies of liquid oral, solutions and
syrup dosage forms. Illustrate your answer with suitable examples. [10]

Preformulation
Preformulation may be described as a phase of the research and development process where the
preformulation scientist characterizes the physical, chemical and mechanical properties of a new drug
substance, in order to develop stable, safe and effective dosage form.
Objectives:
The preformulation investigations confirm that there are no significant barriers to the compound’s
development as a marketed drug. The formulation scientist uses these informations to develop dosage forms.

Principal areas of preformulation


1. Bulk characterization
(i) Crystallinity and polymorphism
(ii) Hygroscopicity
(iii) Fine particle characterization
(iv) Powder flow
2. Solubility analysis
(i) Ionization constant – pKa
(ii) pH solubility profile
(iii) Common ion effect – KSP.
(iv) Thermal effects
(v) Solubilization
(vi) Partition coefficient
(vii) Dissolution
3. Stability Analysis
(i) Stability in toxicology formulation
(ii) Solution stability
– pH stability profile
(iii) Solid state stability
– Bulk stability
– Compatibility

Please go through the details of the Preformulation Chapter of Degree syllabus.


[Ref: Fiese E.F. and Hagen T.A., Preformulation, In The Theory and Practice of Industrial Pharmacy, Ed.
Lachman L, Lieberman H.A., and Kanig J.L.,3rd Edn., p.171–196.]
At this point you should know the following things:
1. Brönsted-Lowry theory of acid and base [Ref: Ionic Equilibria, Physical Pharmacy, Ed. Martin A., 4th
Edn., p.143].
2. Definition of pH.
3. Dynamic equilibrium state.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 2

pKa and Solubility


Ionization of weak acids
The ionization of a weak acid, in water may be written in the Brönsted-Lowry manner as:
k1
HA + H 2O A + H3O+
Base 2 k2 Base 1 Acid 2
Acid 1

This is a reversible process. According to the law of mass action, the velocity or rate of forward reaction, Rf is
proportional to the concentrations of the reactants.
Rf = k1 x [HA] x [H2O]

The velocity of reverse reaction is Rr = k2 x [A–] x [H3O+]


The symbols k1 and k2 are proportionality constants, known as specific rate constant for forward and reverse reaction
respectively.
At equilibrium R f = Rr What is the concentration of water at 250C? Express
in molarity.
or, k1 x [HA] x [H2O] = k2 x [A–] x [H3O+]

or,
[
k1 H 3 O + A
=
][ ] 1000ml H2O has a mass of 997.07g
[see from density chart of H2O]
k 2 [HA][H 2 O ] 997.07
997.07g water = = 55.3moles water
In dilute solution water is in sufficient excess, so [H2O] can be 18.02
taken as constant. Therefore, [H2O] = 55.3 moles/lit = 55.3 M

or,
k1 [ +
[H 2 O ] = H 3 O A ][ ]
k2 [HA]
or, Ka =
[H O ][A ]
3
+
k1
[H 2 O]
where K a =
[HA] k2
Ka is called the ionization constant or dissociation constant or acidity constant of the weak acid HA.

Case 1: For a weak uncharged acid like acetic acid


+
CH3COOH + H2O H3O + CH3COO
At time 0 c 0 0
At time t c-x x x

[ H 3 O + ] [CH 3 COO ]
Therefore, Ka =
[CH 3 COOH ]
x2
=
c x
Since, c>>x the term c–x c
2
Therefore, Ka = x / c or, x = K a c or, [ H 3 O + ] = K a c
Problem: In a liter of a 0.1 M solution, acetic acid was found by conductivity analysis to dissociate into 1.32 x 10–3
gram ions (moles) each of hydrogen ion and acetate ion at 250C. What is the acidity constant, Ka for acetic
acid?

Case 2: For a weak, charged acid like NH4+.


When a salt is formed from a strong acid and a weak base, e.g. NH4Cl, is dissolved in water, it dissociates completely
as follows:
H2O
NH4+Cl NH 4+ + Cl

The Cl– is the conjugate base of the strong acid, HCl, which is 100% ionized in water. Thus Cl– cannot react any
further. According to Bronsted-Lowry system, NH4+ is considered to be a cationic acid and its conjugate base is NH3.

NH4+ + H2O H +
+3O + NH3
Acid 1 Base 2 Acid 2 Base 1
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 3

[ H 3 O + ] [ NH 3 ]
In this case the acidity constant, Ka =
[ NH 4 + ]

Ionization of weak bases


Case1: For non-ionized weak base, B
B + H 2O OH + BH +
Base 1 Acid 2 Base 2 Acid 1
The dissociation constant or basicity constant, Kb, can be expressed as follows:
[OH ] [ BH + ]
Kb = This leads to [OH ] = K b c
[ B]
Case 2: Ionized weak base e.g. Acetate ion
Salts of strong bases and weak acids, such as sodium acetate, dissociates completely in water to give the following ions:

H2O
Na+CH3COO Na+ + CH3COO
+
The Na ion cannot react with water (since it would form NaOH, which is a strong electrolyte and would dissociate
completely into ions.)
The acetate ion is a Bronsted-Lowry weak base, and

CH3COO + H2O OH + CH3COOH


[OH ] [CH 3 COOH ]
Kb =
[CH 3 COO ]
In general, for an anionic base, B–.

B + H 2O OH + HB
[OH ] [ HB]
Kb =
[B ]

Ionization of water
Just like weak acid or weak bases, water ionizes into hydrogen and hydroxyl ions. This auto protolytic reaction is
represented as
H2O + H2O H3O+ + OH
Acid 1 Base 2 Acid 2 Base 1
The law of mass action then applied to give the equilibrium expression:
[ H O + ] [OH ] [ H O + ] [OH ]
K = 3 or, K = 3 or, K [ H 2 O] 2 = [ H 3 O + ] [OH ]
[ H 2 O][ H 2 O] [ H 2 O] 2
Since, H2O is in large excess than H3O+ or OH– hence [H2O] can be takne as constant.
or, K [ H 2 O] 2 = cons tan t = K w
Kw is called dissociation constant, the autoprotolysis constant or the ion-product of water.
The value of Kw is approximately 1 x 10–14 at 250C. It depends strongly upon temperature.
In pure water, the hydrogen and hydroxyl ion concentrations are equal and [H3O+] = [OH–] = 1 x 10–7 M at 250C.
• When an acid is added to pure water the [H3O+] increases and [OH–] decreases but the ion-product of water, Kw,
remains constant.
• Under laboratory condition, distilled water in equilibrium with air contains about 0.03% by volume of CO2
corresponding to a [H3O+] of about 2 x 10–6 M (pH 5.7).
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 4

Relationship between Ka and Kb

HA + H 2O H3O+ + A A + H 2O HA + OH
=
[H O ][A ]
3
+
=
[HA][OH ]
Ka
[HA]
Kb
[A ]
Therefore, K a K b =
[H 3O ][A ] x [HA][OH ]
+
= [H3O+] [OH–] = Kw Therefore, K a K b = K w.
[HA] [A ]
Ionization of polyprotic electrolytes
Acids those donate a single proton and bases those accepts a single proton are called monoprotic electrolytes.
A polyprotic (polybasic) acid is one that is capable of donating two or more protons.
A diprotic (dibasic) acid is one that is capable of donating two or more protons.
An example of diprotic acid is carbonic acid:
+ [ HCO3 ] [ H 3 O + ]
H2CO3 + H2O HCO3 + H3O K1 =
[ H 2 CO3 ]
+ [CO3 2 ] [ H 3 O + ]
HCO3 + H2O CO32 + H3O K2 =
[ HCO3 ]
Example of a triprotic (tribasic) acid is phosphoric acid.

H3PO4 + H2O H3O+ + H2PO4 [ H 3 O + ] [ H 2 PO4 ] 3


K1 = = 7.5 x 10
[ H 3 PO 4 ]
[ H 3 O + ] [ HPO 4 2 ]
H3O+ + HPO 4
2 8
H2PO4 + H2O K2 = = 6.2 x 10
[ H 2 PO 4 ]
[ H 3 O + ] [ PO 4 3 ]
H3O+ + PO4
2 13
HPO 4 + H2O 3 K3 = 2
= 2.1 x 10
[ HPO 4 ]
Each of the species formed by the ionization of a polyprotic acid can also act as a base, thus for phosphoric acid system
the bases are as follows:
3 2 [ HPO4 2 ] [OH ] 2
PO4 + H2O HPO4 + OH K b1 = = 4.8 x 10
[ PO 4 3 ]

2 [ H 2 PO 4 ] [OH ] 7
K b2 = = 1.6 x 10
HPO 4 + H2O H2PO42 + OH [ HPO 4 2
]
[ H 3 PO 4 ] [OH ] 12
K b3 = = 1.3 x 10
H2PO4 + H2O H3PO4 + OH [ H 2 PO4 ]
Therefore K 1 K b1 = K 2 K b 2 = K 3 K b3 = K w

Ampholytes
A species that can function either as an acid or a s a base is called an ampholyte and is said to be amphoteric.
Example: If glycine hydrochloride is dissolved in water, it ionizes as follows:

+ H2O +
Cl H 3NCH2COOH H3NCH2COOH + Cl
+ + +
H3NCH2COOH + H2O H3NCH2COO + H3O
Acid Base
+ +
H3NCH2COO + H2O H2NCH2COO + H3O
Acid Base
H3N+CH2COO– is amphoteric in nature because it can act both as an acid and base with water as follows:
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 5

+
H3NCH2COO + H2O H2NCH2COO + H3O+
Acid Base
+ +
H3NCH2COO + H2O H3NCH2COOH + OH
Base Acid

This type of species having two types of charges on one molecule are called zwitterions. The pH at which zwitterion
concentration is maximum is known as isoelectic point.
Determination of pKa
pKa of a drug molecule can be determined by various methods:
1. Potentiometric (pH) method
2. Spectrophotometric method
3. Partition-coefficient method
4. Conductometric method
5. Solubility method
Among all the methods potentiometric and spectrophotometri cmethods are most popular and accurate methods by
which the pKa are determined.

Potentiometric method of determination of pKa

Step-1: An acid is dissolved in deionized water (100ml). The pH is determined with a previously calibrated pH-meter
(Potentiometer).
Step-2: The solution is titrated with N/10 NaOH solution. Each time 1ml NaOH solution is added.
Step-3: The pH versus volume of NaOH solution added is plotted. A titration curve will be obtained.
Step-4: Where ever the graph rises very steeply that volume is the equilibrium point of the titration curve.
Step-5: The pH is determined from the titration curve where the volume is ½ the volume at equilibrium point.

This method will be sufficient for determining the pKa of a monoprotic acid, but in case of polyprotic acid the
dpH
equilibrium points may not be sharp so in that case the is plotted against V (volume of NaOH solution added).
dV
This first order derivative gives sharp points of equilibrium. From those peaks the equilibrium volumes are determined
V1 V1+ V 2 V 2 +V 3
(say V1, V2, V3 etc). pKa1 = pH at , pKa2 =pH at and pKa3 = .
2 2 2

pH pH

pKa2
pKa
V1/2 V1
Volume of NaOH soln.
Fig. Titration curve of a monoprotic acid pKa1
V1 V2
1 1
V1 (V1+V2)
2 2
Fig. Titration curve of a diprotic acid
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 6

Spectrophotometric method

Whenever a weak acid (HA) is dissolved in water some molecules will be ionized into H+ and A–. If the moiety A
absorbs UV or visible light then there will be two light absorbing species. Each species will have separate mola
absorptivity ( .)
Step-1: Determination of molar absorptivity of HA ( 1).
If solution of HA is prepared in HCl acid then all the molecules will remain in unionized (HA) state. So the
absorbance this solution will be only due to HA species: A1 = 1 b CT. where CT is the total concentration of
the drug n the solution. Where b is the pathlength of the cuvette.
Step-: Determination of molar absorptivity of A– ( 2).
If solution of HA is prepared in NaOH solution then all the molecules will remain in ionized (A–) state. So the
absorbance this solution will be only due to A– species: A2 = 2 b CT. where CT is the total concentration of the
drug in the solution.
Step-3: Determination of absorbance of a solution at any pH in between 2 to 11.
If the solution of the drug is prepared in a pH in between 2 to 11 then the solution will contain both HA and A–
ions but the concentration of each ion may vary. So the absorbance will be contribution of both HA and A–.
Therefore, A = 1 b C1 + 2 b C2 where C1 and C2 are the concentrations of HA and A– respectively in the solution.
Now equations are solved:
A = 1 b C1 + 2 b C2
and CT = C1 + C2.
Solving these two equations the values of C1 and C2 are obtained.
A 1 bC T bC A
C1 = and C2 = 2 T
1b 2b 1b 2b

If Handerson-Hasselbach equation is applied for this monoprotic acid then


[base] C A 1 b CT
pH = pKa + log = pKa + log 1 = pKa + log
[acid ] C2 2 b C T A
A A1
pH = pKa + log
A2 A
In this final equation pH, A, A1, and A2 are known so pKa can be calculated.

N.B. In case of polyprotic acid the number of equations required will be more. For example for a triprotic acid (H3A)
there will be four species: H3A, H2A–, HA2–, and A3–. In this case eight simultaneous equations will be required to
determine the four unknown concentrations (C1, C2, C3 and C4) and four molar absorptivities ( 1, 2, 3, and 4)

A1 = 1 b C1 + 2 b C2 + 3 b C3 + 4 b C4 determined at 1. Eqn. 1
A2 = 1 b C1 + 2 b C2 + 3 b C3 + 4 b C4 determined at 2. Eqn. 2
… ….. ……. ….. ….. …..
A8 = 1 b C1 + 2 b C2 + 3 b C3 + 4 b C4 determined at 8. Eqn. 8

Now these eight equations may be solved by some computer program and the concentration terms are determined.
Putting those terms in each Handerson-Hasselbach equation for two conjugate species the pKa1, pKa2 and pKa3 can
be determined.

Solubility method
S S0
pH = pKa + log
S0
Where S = solubility of a drug at any pH and
S0 = intrinsic solubility of the drug, i.e. solubility of only the unionized form of the drug.
Step-I: Solubility of the drug is determined at a pH where all the molecules of the drug will remain in
unionized state (S0).
Step-: The pH is changed by 1 unit and the solubilities are determined (S).
(S S 0 )
Step-3: pH vs. log is plotted and the slope determined will give the Ka after necessary calculation.
S0
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 7

Significance of pKa

1. From the pKa of a weak acid or weak base the unionized fraction of a drug can be determined at a certain pH. The
unionized fraction has greater absorption rate through any biological membrane. So while designing a dosage form
how a weak acid or weak base will behave in any biological fluid and its rate of absorption and the site of
absorption can be guessed from the pKa. From the table below it is evident that Ibuprofen, a weak acid is absorbed
maximum from stomach and Nitrazepam, a weak base will be absorbed preferrentially from duodenum.

Drug Stomach Plasma Duodenum


PH 1.5 PH = 7.4 PH = 5.0

Weak acid [HA] = 100 [HA] = 100 [HA] = 100


e.g. Ibuprofen
pKa = 4.4

[A--] = 0.13 [A --] = 100,000 [A--] = 398.1

[Total] = 100.13 [Total] = 100,100 [Total] = 498.1

Weak base
e.g. Nitrazepam [B] = 100 [B] = 100 [B] = 100
pKa = 3.2 of its
conjugate acid
[BH +] = 5012 [BH +] = 0.006 [BH +] = 1.6

[Total] = 5112 [Total] = 100.006 [Total] = 101.6

2. The solubility of a weakly acidic and weakly basic drug can be increased if the drug remains in ionized state. So
the solubility vs. pH is plotted to get pH-solubility profile of a drug. While designing a dosage form (oral,
ophthalmic or parenteral) the pH the solution is buffered to that pH where the solubility is maximum and the drug
is reasonably stable.

SOLUBILITY
Definition
The maximum amount of solute that is soluble in one part of solution to make a saturated solution at a certasin
temperature is called the solubility of the drug.

Significance of solubility
1. Increased bioavailability:
• In dealing with a new drug substance, it is extremely important to know something about its solubility
characteristics, especially in aqueous solution, in order to elicit a therapeutic response. Any drug having
solubility less than 10mg/mL in physiologic pH range (pH 1 to 7) will produce bioabsorption problem. A
solubility less than 1mg/mL require salt formation of the drug for better bioavailability.
• When the solubility of a drug cannot be increased by salt formation (e.g. in neutral molecules, glycosides,
alcohols, steroids or where the pKa of a basic drug is less than 3 and the pKa of an acidic drug is more than
10) then the drug is dissolved with a cosolvent and filled in a soft gelatin capsule.
• Griseofulvin, an antifungal drug, when given orally the absorption is very less. So it is given with fat meal.
The rate of dissolution rate of griseofulvin is increased by micronization (in a fluid energy mill) or by solid
dispersion technique to increase its oral bioavailability.
2. Taste masking: Chloramphenicol is very bitter in taste so it is very difficult to make a paediatric liquid dosage form
with chloramphenicol base. Chloramphenicol palmitate is taken, the solubility of which is very low compared to
chloramphenicol base. When a suspension is prepared due to its low solubility it does not produce any bitter taste.
3. Reducing degradation in the GIT: Drugs like erythromycin will degrade while passing through the acid
environment of stomach, so erythromycin is delivered as erythromycin proprionate or estolate while preparing
paediatric suspension. This solubility of these esters are very less in acidic pH. Thus they are saved from gastric
pH.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 8

Determination of solubility of the drug


Step-1:
Method A: Some excess amount of drug is dissolved in 10ml of solvent. The suspension is shaken overnight (24hrs) in
a fixed temperature water bath.
Method B: Some excess amount of drug is dissolved in 10ml of solvent by heating, then the suspension is put in a fixed
temperature water bath.
Step-2: The solids are separated from saturated solution either by filtration through membrane or by centrifugation.
Step-3: The filtrate (or supernatant liquid after centrifugation) is assayed to determine the solubility of the drug. The
assay method may be gravimetric, UV-spectrophotometric, HPLC etc.

Intrinsic solubility of a drug (S0)


This is the fundamental solubility of a drug when it is completely unionized.
• For a weak acid the intrinsic solubility is the solubility of the drug determined in a strongly acidic solution.
• For a weak base the intrinsic solubility is the solubility of the drug determined in a strongly alkaline solution.
• For a non-ionic molecule there will be no measurable change in the solubility in either acidic or alkaline solution.

In case of weak acid and weak base the solubility can be manipulated by changing the pH of the solution.
In case of non-ionizable molecules the solubility can be manipulated either by changing the solvent, or by addition of
cosolvent or by complexation.

Approaches of increasing the solubility of drugs


1. By changing the pH of the solution
For a weak acid the relationship between the pH of the solution and the solubility of the drug is:
S S0
pH = pKa + log
S0
where S = overall solubility of the drug = Concentration of ionized fraction + Concentration of unionized fraction (Su)
For a weak base (BH+) the relationship between the pH of the solution and the solubility of the drug is:
S0
pH = pKa + log
S S0
So in case of a weakly acidic drug the solubility can be increased by increasing the pH and for a weakly basic drug the
solubility can be increased by decreasing the pH.

2. By changing the solvent


• The first preference of solvent is water. If the solubility is very less in water then water may be replaced, either
partially or completely, with one or more water-soluble solvents like ethanol, glycerol, sorbitol, propylene glycol
etc. The solvents are called cosolvents, and the phenomenon as cosolvency. These types of non-toxic cosolvents
are used in designing oral liquid dosage forms.
Example:
1g Phenobarbital is soluble in 1000ml water, in 10ml alcohol.
It is found that solubility of phenobarbital is 1.5%w/v in a solvent mixture where alcohol is 22%, glycerin is
40% and the remainder is water (i.e. 38%)

• In case of assaying a drug by chromatography or spectrophotometry any solvent in which the drug is adequately
soluble may be taken. For example aqueous methanol for HPLC, chloroform in spectrophotometry, etc.

3. By changing the polymorphs


• Whenever a drug is crystallized from some solvent, depending on the conditions of crystallization, the
polymorphic shape is changed. For example, if cooled very quickly then metastable polymorphs will be formed
and if cooled very slowly then stable crystalls will form. The metastable form has higher solubility that the stable
polymorph.
• While crystallization solvent molecules may be entrapped within the crystal lattice in stochiometric ratio – these
types of crystals are called solvates. If the solvent molecule entrapped are water (H2O) molecules then the crystals
will be called hydrates. The solubility of these pseudopolymorphs may be arranged in ascending order:
Hydrates < Anhydrous < Solvates
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 9

4. By adding a suitable surfactant


A sufractant when dissolved in water in a concentration over the critical micelle concentration (CMC) will produce
micelles. The drug, both ionized and unionized forms, will partition between water and micelle. If the concentration
of surfactant is increased over CMC the partition of the drug into the micelle will increase which will show an
apparent increase of solubility of the drug.
Sodium lauryl sulfate, a surfactant, increases the solubility of benzoic acid.
In case of oral liquid dosage forms generally non-ionized surfactants are used (e.g polysorbate 80 i.e. Tween80) to
increase the solubility of a drug.
5. By complexation
Caffeine increase the solubility of benzoic acid by forming a water-soluble complex. Solubility of para
aminobenzoic acid (PABA) can be increased by complexing with caffeine.

Approaches of decreasing the solubility of drugs


1. By esterification:
The soubility of chlopramphenicol can be decrease by forming its ester with palmitic acid.
2. By coating with polymers
Drug particles may be coated with ethylcellulose to retard its water solubility. Cellulose acetate phthalate (CAP),
hydroxypropylmethylcellulose phthalate (HPMCP) etc. polymers reduce the solubility of drug particles in the acid
medium of stomach.
3. By changing the polymorph
Stable polymorphs have lower aqueous solubility than the metastable forms. So by changing the condition of
crystallization stable polymorphs may be produced.
4. By selecting the hydrated forms
Anhydrous ampicillin has greater water solubility than ampicillin-trihydrate. In anhydrous forms the drug powder
has an inherent demand for water, hence its solubility is higher than the hydrates where the demand for water is
satisfied.

Example: Calculating of solubility of a weak acid by changing the pH of the solution.


Sodium phenobarbitone is a salt of weak
acid. When this salt is dissolved in water it H H
O N ONa O N O
readily dissociates into its ionized form (P–)
H2O
2 5 CH N C2H5 N + Na+
Determination of intrinsic solubility (S0)
The solution of the salt is made acidic with O O
HCl acid. In this acidic pH all the
phenobarbitone will remain in unionized Sodium phenobarbitone Ionized form
(HP) state. Some molecules will precipitate H H
(HPsolid) as solid and the solution will be a O N OH O N O
saturated solution of unionized form (HPsol).
The concentration of the supernatant C2H5 N + H2O C2H5 N + H3O+
saturated solution is the intrinsic solubility
of the drug (S0). O O
--
Weak acid (HP) Ionized form (P )
Determination of solubility
The equilibrium equations of this system are:

HP solid HP sol
HP sol + H20 H3O+ + P
Since the concentration of the unionized form in solution [HP]sol is constant, so S0 = [HP]sol and the constant for acid-
[H O + ][P ] [ HP]
base equilibrium, equation is: K a = 3 or, [ P ] = K a
[ HP ] [H 3O + ]
The total solubility S of phenobarbital will be:
S0 Ka
S = [HP] + [P–] or, S = S 0 + K a or, S = S0 1+
[ H 3O ]+
[H 3O + ]
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 10

Relationship between solubility and pH of a solution


Determination of Ka of a weak acid or base from the solubility at various pH
S0 S S0 Ka S S0
S = S0 + Ka +
or, = +
or, log = log K a log[ H 3 0 + ]
[ H 3O ] S 0 [H 3O ] S0
S S0 S S0
or, log[ H 3 0 + ] = log K a + log or, pH = pK a + log
S0 S0
Keeping sodium penobarbitone salt in strong acid the intrinsic solubility (S0) is determined. Then at any pH the
solubility (S) of phenobarbitone is determined. If pH, S and S0 is known then Ka can be calculated using the above
equation.

PARTITION COEFFICIENT

If a drug is added to the immiscible solvents, the drug will be distributed between two layers. At equilibrium the ratio
of concetration of drug in two layers will be constant.
C
K= 1 where C1 = concentration of drug in liquid-1.
C2
C2 = concentration of drug in liquid-2.
K = distribution coefficient
The equilibrium constant K is known as the distribution ratio, distribution coefficient, or partition coefficient.

N.B. This equation is true when the solution is a dilute one. In concentrated solution the activity should be determined.
a C
i.e. K = 1 = 1 1
a2 2C 2

Applications of partition coefficient

1. Effect of partition coefficient on ionic dissociation and molecular association

The solute may exist partly or wholly as associated


molecules in one of the phases or it may dissociate
H O
into ions in either of the liquid phases. The O
C
distribution law applies only to the concentration of C
the species common to both phases. O H O
In benzoic acid system it is undissociated benzoic
acid, which is the common species. Oil (HA) 2
[ HA] o
K= Water O O
[ HA] w
C C + H+
Example: In peanut oil benzoic acid remains in free
O H O
monomer state. In benzene benzoic acid remains in
dimer state. Here two molecules of benzoic acids are
HA A
bound by hydrogen bonds.
In practice the apparent partition coefficient (K’ ) is
determined.

Co [ HA] O + [( HA) n ] O
K' = =
CW [ HA]W + [ A ]W

2. Extraction

To determine the efficiency with which one solvent can extract a compound from a second solvent the value of K is
required in both solvents. This extraction principle is required in
• analytical chemistry to develop the solvent system of TLC, HPLC and HPTLC
• in synthetic chemistry to purify a drug and
• in separating the active constituents from crude drug extract of plants or animals.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 11

concentration of solute in original solvent (C1 )


Partition coefficient in two immiscible liquids, K =
concentration of solute in extracting solvetn (C 2 )
Solvent-1 Solvent-2 K
Solvent 1
V1 V2
(Original
Initial mass V1 solvent)
of solute W0 0
1st W1 / V1 KV1 Solvent 2
K= W1 = W0 V2
Extraction W1 W0 – W1 (W0 W1 ) / V 2 KV1 + V 2 (Extracting
solvent)
2nd W 2 / V1 2
K= KV1
Extraction W2 W1 – W2 (W1 W 2 ) / V 2 W 2 = W0
KV1 + V 2
3rd W3 / V1 3
K= KV1
Extraction W3 W2 – W3 (W 2 W3 ) / V 2 W3 = W 0
KV1 + V 2

In case of extraction of a costly drug from its crude drug e.g. vincristine or vinblastine from Vinca rosea the minimum
number of extraction steps (n) is large.
In case of a low cost drug if the number of extraction steps are large then additional cost due to each extraction step
(like the cost of solvent, energy or manpower) will increase which will make the process uneconomical. So by
determining the partitioning effect (K) in the two solvents and from the above equations the minimum theoretical
number of extraction steps are calculated.

3. Preservative action of weak acids in emulsion

Enzymes produced by yeasts, molds and bacteria must be destroyed or inhibited to prevent deterioration of product.
Example: Sodium benzoate is frequently used as preservative, especially in food and oral products. The preservative
action of sodium benzoate is due to unionized benzoic acid only.
The MIC90 of benzoic acid = 25 mg/100ml.
This is probably due to greater penetration of unionized acid (HA) through the biological membranes of
microorganisms.
Microorganisms generally resides in aqueous phase and oil-water interface. So the concentration of [HA] > MIC90 of
the benzoic acid. Benzoic acid will be distributed into oil and water phases. So for determining the preservative
concentration of sodium benzoate calculations must be carried out to keep [HA] > MIC90 in the aqueous phase.

4. Drug action and partition coefficient

The partition coefficient of a drug is an indication of the lipophilic and hydrophilic character of a drug molecule.
For example a lipophilic sedative drug can cross the blood-brain-barrier quickly and thus it will produce quick onset of
action. But a very lipophilic drug is difficult to be carried from the site of absorption, through blood (aqueous), to brain.
So the molecule must have optimum hydrophilicity also so that it can be carried by the aqueous blood.

During preformulation studies partition coefficients in various oil phases correlates well with the permeation of various
biological membranes.
Example:
Oil phase Biological membrane
n-butanol Buccal
n-pentanol
n-octanol
Ethyl acetate Gastrointestinal
Ether
Oleyl alcohol
Toluene
Cyclohexane Blood brain barrier
Heptane
Carbontetrachloride
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 12

CRYSTAL MORPHOLOGY, POLYMORPHISM, HYDRATES & SOLVATES


Crystal
Crystals are characterized by repetition of atoms, molecules or ions in a regular three-dimensional structure.

Crystal Habit
Crystals may have different external structures (outer appearance) with a single internal structure. This external
structures are known as crystal habits.
Crystal habits are influenced by the environment of the growing crystals. Five types of crystal habits are generally
recognized during crystallization.
(i) Platy: plates, crystal growth in length and width and moderate height.
(ii) Acicular: needle-like, crystal growth in one direction.
(iii) Tabular: moderate expansion of two parallel faces.
(iv) Bladed: flat acicular
(v) Prismatic: columns.

Factors affecting crystal habit

1. Excessive super saturation tends to transform a prism to a needle shape.


2. Cooling rate and agitation changes habit as it changes the degree of supersaturation, e.g. naphthalene gives thin
plates (platy) if rapidl;y recrystallized in cold ethanol or methanol, whereas slow evaporation yields prisms.
3. The crystallising solvent affects habit by preferrential absorption on to certain faces, inhibitingtheir growth.
Resorcinol produces needles from benzene and squat prisms from butyl acetate.
4. The addition of cosolvents or other solutes and ions which change habit by poisoning crystal growth in one or
more directions. Sodium chloride is usually cubic, but urea produces an octahedral habit.

Characterization of crystal morphology


1. Optical microscopy
2. Scanning Electron Micorscopy (SEM)

Polymorphism
When crystals exists in more than one internal structure (i.e. packing pattern) the various crystalline forms are called
polymorphs and the phenomenon is known as polymorphism.
Depending on the thermodynamic stability, the polymorphs are divided into three categories stable, metastable and
unstable.
Unstable form has a tendency to transform into stable form. Metastable forms in dry state will remain stable, but if
melted or dissolved will form stable polymorph.

Characteristics of polymorphs

Characteristics Stable polymorph Metastable polymorph Unstable polymorph


Packing of molecules in crystal lattice Tightly packed Less tightly packed Loosely packed
Melting point Highest Moderate Lowest
Rate of dissolution Lowest Moderate Highest

Polymorphism and bioavailability


Many drugs are hydrophobic and have very limited solubility in water. If the drug remains in several polymorphic
forms then the stable one will produce the slowest rate of dissolution and it may show minimum bioavailability.
For highly water soluble drugs polymorphism does not show any problem in dissolution rate.

Example: Chloramphenicol palmitate has three polymorphs (stable), (metastable) and (unstable). When
chloramphenicol palmitate suspension is prepared from or polymorph it is found that bioavailabilty is higher with
the metastable form.
Example: Two polymorphs of aspirin can be obtained by recrystallization of aspirin from 95% ethanol or n-hexane. The
polymorph obtained from n-hexane is found to have greater solubility in water than the polymorph obtained from
ethanol.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 13

Polymorphism and melting point of cocoa butter suppositories

Theobroma oil or cocoa butter suppositories are meant to be melted at body temperature 370C but should remain as
solid at room temperature during storage period.
Cocoa butter is available in three polymorphs– (m.p. 200C), (m.p.360C) and (m.p. 150C). -form is the metastable
form, -form is the stable form and -form is the unstable form. During melting of cocoa butter by fusion method the
following phenomena are found:

Procedure Observation Explanation


The cocoa butter is melted The suppositories melts below If melted at high temperature and cooled very
applying high temperature 300C i.e. at room temperature it quickly then the molecules form the -crystals in
(600C) and then quickly will melt. So it will be difficult more amount so the suppositories melt below
chilled. to handle at room temperature. 300C. The metastable -form will revert to stable
-form, but it will take several days for this
procedure.
The cocoa butter is melted at The suppositories do not melt at The use of low temperature and slow cooling rate
low temperature (40 to 500C) room temperature. The melting allows direct formation of -crystals having a
and then cooled slowly. point will be above 360C . melting point of 360C.

So during preparation of cocoa butter suppositories 2/3rd portion of cocoa-butter base is melted and then the container is
removed from the heat source. The rest of the base is melted by stirring only without application of heat.

Polymorphism and caking of suspension

In case of a suspension the particles will sediment below and the particles will come closer to each other. During a long
storage life the suspension may experience several cycles of temperature change. During the hot period the metastable
form will get dissolved in the stagnant layer and during the cool period the particles will grow and crystal bridges may
form with the stable crystals. These crystal-bridges will give rise to irreversible caking of suspension.

Amorphous form
If a solid does not have any fixed internal structure that form is called amorphous form. They have molecules randomly
placed as in a liquid. E.g. Amorphous novobiocin.
Characteristics of amorphous forms:
Melting point: Stable > metastable form > unstable form > amorphous form of the same drug
Dissolution rate: Stable < metastable form < unstable form < amorphous form of the same drug

Preparation
Amorphous forms are prepared by rapid precipitation, lyophillization or rapid cooling of molten liquids e.g. glass

Glass transition temperature, Tg


Tg is a characteristics tempeatuer of amorphous form. Below Tg the amorphous form will be brittle and is described as
glassy state. Above Tg the solid becomes plastic or rubbery. So Tg is the minimum temperature at which the solid
becomes amorphous (plastic) from glassy state.
Application:
1. Tg can be reduced by addition of plasticizers. Plasticizer molecules, either, disturb or distort the molecular
arrangements, thus they reduce the Tg.
2. During milling, all the solids must remain below Tg.
3. Amorphous novobiocin is more soluble and has higher bioavailability than its crystalline form.

Polymeric materials
Polymers are very large molecules. They are so large and flexible that it is not possible for them to align perfectly to
form crystals. They usually have ordered regions within the structure, surrounded by disorder, so they are described as
semicrystalline. The degree of crystallinity depends up on processing conditions.

Methods of characterization of polymorphs


1. Hot stage microscopy, 2. Differential Thermal Analysis, 3. Differential Scanning Calorimetry
4. Thermogravimetric Analysis (TGA) 5. X-ray powder diffraction 6. IR-Spectroscopy
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 14

Molecular Adducts
During the process of crystallization, some compounds have a tendency to trap the solvent molecules.

1. Non-Stoichiometric inclusion compounds (or adducts)


In these crystals solvent molecules are entrapped within the crystal lattice and the number of solvent molecules are
not included in stoichiometric number. Depending on the shape they are of three types :-
(1) Channel
When the crystal contains continuous channels in which the solvent molecule can be included. e.g . Urea
forms channel.
(2) Layers:- Here solvent molecules are entrapped in between layers of crystals.
(3) Clathrates(Cage):- Solvent molecules are entrapped within the cavity of the crystal from all sides.

2. Stoichiometric inclusion compounds (or stoichiometric adducts)

This molecular complex has incorporated the crystallizing solvent molecules into specific sites within the crystal lattice
and has stoichiometric number of solvent molecules complexed.
When the incorporated solvent is water, the complex is called hydrates and when the solvent is other than water, the
complex is called solvates. Depending on the ratio of water molecules within a complex the following nomenclature is
followed.
(i) Anhydrous : 1 mole compound + 0 mole water
(ii) Hemihydrate: 1 mole compound + ½ mole water
(iii) Monohydrate: 1 mole compound + 1 mole water
(iv) Dihydrate : 1 mole compound + 2 moles water

Properties of solvates / hydrates


(i) Generally, the anhydrous form of a drug has greater aqueous solubility than its hydrates. This is because the
hydrates are already in equilibrium with water and therefore have less demand for water. e.g. anhydrous forms
of theophyline and ampicillin have higher aqueous solubility than the hydrates.
(ii) Non aqueous solvates have greater aqueous solubility than the non-solvates. e.g. chloroform solvates of
griseofulvin are more water soluble than their nonsolvate forms.

FLOW RATE

Application of free flowing powders in pharmacy

1. Powders are required to produce tablets or capsules. Free flowing powders flow uniformly into the die cavity of
tablet punching machine and inside the empty gelatin shells. A free flowing powder produces uniform content of
drug in the tablets and capsules.
2. Free flowing powders show reproducible filling of tablet dies and capsules dosators, which improve weight
uniformity and physicomechanical properties (e.g. hardness).
3. Poor powder flow can result in excess entrapped air within powders which in some high-speed tabletting
conditions may promote capping or laminations.
4. Poor powder flow can result from excess fine particles in a powder, which increase friction in between particle and
die wall. It may cause lubrication problem.
5. In industry powders are required to flow from one location to another and this is achieved by different methods,
such as gravity feeding, fluidization in gases and liquids and hydraulic transefer. In each of these examples
powders are required to flow.
6. In capsule filling machine, especially Lily type capsule filling machine the powder must be free flowing to
uniformly fill the base of the capsule shells. In case of Zanasi-type filling machine cohesive powders are required.

Characterization of powder flow:

Indirect methods Direct methods


(a) Angle of repose (a) Hopper flow rate
(b) Shear cell determinations (b) Recording flow-meter
(c) Bulk density measurements
(d) Critical orifice diameter.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 15

Angle of repose

Angle of repose of a powder sample is a parameter that shows interparticle cohesion.


If cohesion between powder particles is less then powder flow-property is better. Due to cohesion, powder experiences
a drag force against flow.
Interparticle cohesive forces are due to
(i) non-specific van der Waals forces. It increases as particle size decreases and moistue content increases.
(ii) Surface tenbsion forces between adsorbed liquid layers at the particle surface and
(iii) Electrostatic forces arising from contact or friction with the wall of the equipment.

Methods of determination of angle of repose ( )

Apparatus Method Angle defined


Fixed height Angle of repose

Fixed base cone Angle of repose

d
Tilting table Angle of repose

Rotating cylinder Dynamic angle of repose

Ledge Drained angle of repose

Crater Drained angle of repose

Platform Drained angle of repose


BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 16

Observations
Same powder may produce different angle of repose due to
(a) different methods of determination
(b) different ways the samples were handled prior to measurement.

Interpretation
As a general guide > 500 unsatisfactory powder flow
< 250 very good powder flow
250 < < 500 satisfactory flow

Shear cell determination Normal load


A shear cell is a simple instrument that consists of two halves. Shearing Movable
Powder is filled inside the cell. Weights are placed on the lid of force uper half
the assembled cell to apply normal pressure ( ). The lid is Pulley
connected by a pulley and a string to a weight to apply shear Fixed
stress ( ). lower half
Shear stress is slowly increased, at a certain shear stress ( ) the Shear plane
lid will move over the fixed lower half.

Observations
It is found that if is increased then also increases. Generally the plot of versus is called Mohr diagram.
normal force
Normal stress ( ) =
cross sec tional area of powder Normal
normal shear force stress
Shear stress ( ) =
cross sec tional area of powder te
Mohr diagram
Slope = tan = coefficient of friction a Shear
= angle of internal friction stress
a = apparent tensile strength
Fig. Mohr diagram
te = cohesion coefficient.

Interpretation of Mohr diagram

Property Powder characteristics


tan Greater slope better flowability
a , te When powder is non-cohesive the
yield locus (line) passes through
origin
i.e. te = 0 and a = 0

Bulk density measurement


Procedure:
A powder mass, M, is taken in a measuring cylinder. Tapped 1000 time with a mechanical tapping device. At
equilibrium there will be no change in height of the powder bed At this condition it is said that the powder bed is
consolidated. The initial volume V0 and the final tapped volume Vf are determined.
M M
Initial density, D0 = and final density (or tapped density), D f =
V0 Vf
Df
Hausner ratio
D0
Significance: It is related to interparticle friction. So it can be used to predict powder flow properties.
For coarse, free flowing powders the Hausner ratio is approximately 1.2.
For more cohesive, less ree-flowing powder (e.g. flakes) will have a Hausner ration greater than 1.6.
Interpretation: Greater the Hausner ratio more cohesive will be the powder and flowability will be reduced.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 17

% compressibility (by Carr)


Df D0 V0 V f
% compressibility = x 100 = x 100
Df V0
Interpretation

% compressibility Flow description Examples


5 – 15 Excellent Free flowing granules
12 – 16 Good Free flowing powdered granules
18 – 21 Fair Powdered granules
23 – 28 Poor Very fluid powders
28 – 35 Poor Fluid cohesive powders
35 – 38 Very poor Fluid cohesive powders
> 40 Extremely poor Cohesive powder
So if % compressibility is high then the powder is more cohesive and less free-flowing.

Critical orifice diameter


Powder is filled in a shallow tray up to a uniform depth with near Shallow tray
uniform packing. The base of the tray is perforated with a series of
Powder
holes having gradually increasing diameter, which are blocked either
by resting the tray on a plane surface or by the presence of a simple
shutter. Orifices
The critical orifice diameter is the size of the smallest hole through
which powder just discharges when the tray is lifted or the shutter is removed.
Critical orifice diameter is a direct measurement of powder cohesion and arch strength and is related by the equation:
r
tan = when r = particle radius and x = critical orifice diameter
x

Direct methods

Hopper flow rate


It measures the rate at shich powder discharges from a hopper. A simple shutter is placed under the hopper outlet. The
hopper is filled with powder. The shutter is then removed and the time taken for the powder to discharge completely is
recorded.
Amount of powder disch arg ed
Flow rate =
Time taken for complete emptying
Discharge tube of different diameters are selected to provide a good model for a particular flow application.

Recording flow-meter
Powder flows from a hopper on to a balance. In analogue balance a chart Fig. 13
recorder is used to produce a permanent record of increase in powder mass
with time. In digital balance the flow rate is measured with time.
g/sec

Improvement of powder flowability time

Powder Nozzle Plotter


1. Alteration of particle size and size distribution
Coarse particles less cohesion better flowability.
Fine particles greater cohesion poor flowability
In a powder sample if proportion of fines are large greater cohesion
poor flowability.

Remedy:
• Granules are prepared for making coarse particles. Granules are Digital Balance
prepared for making coarse particles.
• Granules are sieved to get particles of uniform size distribution.
• During lubrication the granules are mixed gently to reduce the fines.
BPUT / M.Pharm. Sem-1 / Chapter-1 / Preformulation Studies / A. Samanta 18

2. Alteration of particle shape and texture


Shape: Spherical particle good flow prperty
Irregular particles more mechanical interlocking

Texture
Very rough surface more cohesive interlocking.
Smooth surface particles less cohesive good flow

Remedies:
(i) Acicular particles can be made spherical by temperature-cycling crystallization.
(ii) Spray-drying produces near spherical particles.
(iii) Solvent is sprayed on a bed of coarse particles while the granules are rotated inside a pan-coater. The sharp
edges will be smoothed or rounded.
(iv) Texture may be altered by controlling the production method, for example crystallization condition.

3. Alteration of surface force


Electrostatic forces
Static charge may develop if the powder particles are in close contact of the wall of the instruments or are moving at
very high velocity through an instrument. If charges develop on the particle surface the flowability of the powder will
decrease.
If the granules or powder become very dry the probability of charge production will increase. So there must be residual
moisture (at least 4% moisture) on the particle surface.
Remedies:
1. Frictional contacts are reduced, for example powder is poured down chutes or conveyed along pipes
pneumatically, the speed and length of transportation should be minimized.
2. Proper earthing of equipments.

Moisture content
Adsorbed surface moisture films tends to increase bulk density and reduce porosity.
Remedies:
1. If excessive moisture content is present the powder is dried (up to 4% moisture content).
2. If the powder has hygroscopicity then it is stored and processed in low-humidity conditions.
3. If the powder is excessively dried then static charge will develop on the powder particles in that case moisture is
sprayed and mixed gently.

4. Formulation additives: flow activators


Flow activators are substances those reduces adhesion and cohesion of the powder.
Example:
1. Glidants like talc, corn starch, magnesium stearate reduce the electrostatic interaction in between the particles and
equipment wall.
2. Colloidal silica gel has very high specific surface area. It reduces the bulk density of tightly packed powder. If
excessive amount of colloidal silica gel is added then flooding will occur.
3. If the powder contains excessive moisture then magnesium oxide is added as flow activator. The particles of
magnesium oxide disrupts the continuous film of adsorbed water.
4. Silicone treated powders like silicone-coated-talc or sodium bicarbonate is used to improve moist or hygroscopic
powders.

5. Alteration of process conditions


Use of vibration-assisted hoppers
If arching or bridging occurs within a hopper, the stress is increased by vibrating the hopper mechanically. Both
amplitude and frequency of vibration can be altered. E.g. compressed air-device or hammer to high frequencies
produced.
Use of force feeders
Fitting of vibrating baffles at the bottom of the hopper forces the powder into the die or capsule shells.