Analysis of the complex

The analysis of complex involves:

➢ Determination of the Stoichiometric ratio
of ligand to acceptor
➢ The stability constant of the complex.

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 Analysis of the complex
Method of continuous variation

◼ In this method an additive property such as

spectrophotometric extinction coefficient, refractive
index or dielectric constant can be used.
◼ When there is no complexation between these
species (A and B), the value of property is additive.
◼ In case of complexation, these properties change,
i.e. additive phenomena do not hold good.

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Spectrophotometric approach

3
A B Mole Absorb. In Absorb. Of the Abs differ.
fraction of absence of complex
A complex

0 10 0 0.1

1 9 0.1 0.12

2 8 0.2 0.3

3 7 0.3 0.34

4 6 0.4 0.36

5 5 0.5 0.4

6 4 0.6 0.45
7 3 0.7 0.41

8 2 0.8 0.3

9 1 0.9 0.21

10 0 1 0.11

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 Analysis of the complex
Method of continuous variation
◼ A plot of
absorbance
difference
versus mol
fraction
◼ (from Martin,
Physical
Pharmacy, p 261)

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 Analysis of the complex
Method of continuous variation
◼ If the magnitude of the measured property (e.g. A) is
proportional only to the concentration of the complex
(MAn ) the molar ratio of ligand A to the metal M can be
determined.
◼ The concentration of M is held constant and that of A is
varied and MAn can be determined by spectroscopic
analysis.

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Analysis of the complex
Method of continuous variation
◼ The equation of complexation is:
◼ M + nA = MAn
◼ Where n is the number of ligand molecules
◼ The Stability constant (K) is:
◼ K = [MAn]/[M][A]n
◼ Log [MAn] = log K + log [M] + n log [A]

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Analysis of the complex
Method of continuous variation

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 Analysis of the complex
Solubility method
◼ Phase–solubility diagram is constructed to determine not
only the value of the stability constant but also to give
insight into the stoichiometry of the equilibrium.
◼ Experimentally, an excess of a poorly water-soluble
drug (i.e. a substrate or drug, D) is introduced into
several vials to which a constant volume of an aqueous
vehicle containing successively increasing
concentrations of the CD.
◼ The vials are agitated at constant temperature until
equilibrium.
◼ The suspensions are then filtered and the total
concentration of the drug (Dt) determined based on
appropriate analytical techniques. 9
 Analysis of the complex
Solubility method
◼ The phase–solubility profile is then constructed
by assessing the effect of the CD on the
apparent solubility of the drug (D).
◼ Based on the shape of the generated phase–
solubility relationships, several types of
behaviors can be identified.
◼ Phase–solubility diagrams fall into two major
types: A and B.

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 Analysis of the complex
Solubility method

◼ A-type profiles.
◼ In A systems, the apparent
solubility of the substrate
increases as a function of CD
conc.
◼ Three subtypes have been
defined:
◼ 1- AL profiles indicate a linear
increase in the solubility as a
function of CD conc.

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 Analysis of the complex
Solubility method

◼ A-type profiles.
◼ 2- AP systems indicate an
isotherm where the curve deviates
in a positive direction from
linearity (i.e. the solubilizer
becomes more effective at higher
conc.).
◼ 3- AN relationships indicate
a –ve deviation from linearity
(i.e. the solubilizer becomes less
effective at higher conc.).
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 Analysis of the complex
Solubility method
◼ A-type profiles.
◼ Taken as a whole, these isotherms indicate that water soluble
complexes are being formed with solubilities higher than that of
the uncomplexed substrate.
◼ AL-type relationships are first order with respect to the CD and
may be first or higher order with respect to the drug (i.e. D•CD,
D2•CD, D3•CD, etc).
◼ If the slope of the AL isotherm is greater than unity, higher order
complexes are assumed to be involved in solubilization.
◼ Although a slope of less than one does not exclude the
occurrence of higher order complexes, a one-to-one complex is
often assumed in the absence of other information.

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 Analysis of the complex
Solubility method
◼ A-type profiles.
◼ AP systems suggest the formation of higher order complexes with
respect to the CD at higher CD concentrations (i.e. D•CD2, D•CD3,
etc).
◼ The stoichiometry of the formed complexes may be implied by the
extent of curvature of the phase–solubility profile. Thus, an
isotherm best fit to a quadratic function suggests the formation of a
one-to-two (D•CD2) complex, one best fit to a cubic function
suggests a one-to-three complex (D•CD3), and so on.

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 Analysis of the complex
Solubility method
◼ A-type profiles.
◼ AN profiles have several explanations including bulk
changes imparted to the solvent by the solubilizer at
various concentrations (i.e. the solubilizer is altering the
bulk properties of the media by changing its viscosity,
surface tension or conductivity) and/or self-association of
the solubilizer at high concentrations.

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Solubility method:
A-type profiles-Equilibrium constants

◼ Do = intrinsic solubility
◼ CE = complexation efficiency
◼ K1:1 = stability constant

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 Analysis of the complex
Solubility method
◼ B-type profiles.
◼ These indicate the formation of
complexes with limited water
solubility and are traditionally
observed with naturally occurring
CDs, especially βCD.
◼ Two subclasses have been
described including BS and BI
systems.

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 Analysis of the complex
Solubility method
◼ BS -type profiles.
◼ As CD concentration increases, a soluble complex
forms which increases the total solubility of the
substrate.
◼ At a particular point, the drug maximum solubility is
achieved which is the sum of Do plus any drug-CD
complex.
◼ Additional CD generates additional complex which
precipitates but so long as solid drug remains,
dissolution and complexation can occur to maintain
the value of Dt.
◼ After consuming the solid drug, addition of more
CD results in formation of additional insoluble
inclusion complex which precipitates and further
depletes the total drug concentration.

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 Analysis of the complex
Solubility method

◼ BI -type profiles.
◼ Similar to BS profiles except
that the formed complexes
are so insoluble that they do
not give rise to the initial
ascending component of the
isotherm.

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