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Adsorption and Partition Studies of Fluconzole

Article May 2009

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Ashok A. Hajare

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Asian J. Research Chem. 2(2): April.-June, 2009

,

www.ajrconline.org

ISSN 0974-4169
RESEARCH ARTICLE

Adsorption and Partition Studies of Fluconzole

Ashok A. Hajare*, Mahesh N. Mali, Sushil Sarvagod, Sachin Kurane, Shweta Patwardhan and Arun S.
Dange
Bharati Vidyapeeth College of Pharmacy, Kolhapur-416013, M.S., India
*Corresponding Author E-mail: aahajare@rediffmail.com

ABSTRACT

The present work was aimed to study the partition coefficient and adsorption of Fluconazole. Fluconazole is an
antifungal drug used in treatment of superficial and systemic fungal infections. It undergoes extensive hepatic first pass
metabolism. Adsorption studies were performed on hydrophobic adsorptives viz. activated charcoal and Talc I. P. at
temperatures 25, 35 and 45C. The partition studies were carried using different lipophilic solvents viz.
dichloromethane, dichloroethane, hexanol and n-octanol at constant temperature of 25C. The partition coefficient was
determined by shake flask method. Adsorption studies on talc and activated charcoal follow both the Freundlich and the
Langmuir adsorption isotherms suggesting possibility of mono as well as multilayer physical adsorption revealing poor
oral bioavailability. Partition studies indicates that increase in carbon chain length of lipophilic solvent decreases
partition coefficient of the drug. Closeness of partition coefficient values to unity suggests possibility of drug to cross
the biological membrane of the microorganism to act locally. From adsorption and partition study it can be presumed
that Fluconazole may be formulated in topical dosage form for local effects to improve its therapeutic actions.

KEY WORDS: Fluconazole, adsorption, adsorption isotherms, partition coefficient

1. INTRODUCTION:

Fluconazole an antifungal agent applied topically for its

action against the fungal infection in the form of
semisolid formulation. It is commonly marketed under
the trade name Diflucan or Trican. The necessity of
physical characterization of drug and its suitability for
particular type of formulation is must for its
development.
Partition coefficient (P) is the ratio of concentrations of
unionized compound in the two phases of a mixture of
immiscible solvents at equilibrium. It is expressed as
K
= Co / Cw
.. (1)
Where, K is partition coefficient, Co is concentration of
solute in organic phase and Cw is concentration of solute
in aqueous phase. Partition coefficient is a measure of
differential solubility of the compound between two
solvents1. For ionizable substances measurements are
made only on their non-ionized form (free acid or free
base) produced by the use of an appropriate buffer with
a pH of at least 1 pH unit below (free acid) or above
(free base) the pKa. Usually solvent pair chosen is
aqueous (water) and hydrophobic (n-octanol).

Received on 26.03.2009
Accepted on 22.05.2009

Modified on 10.04.2009
AJRC All right reserved

Asian J. Research Chem. 2(2): April.-June, 2009 page 213-219

Partition coefficients are useful in estimating distribution

of drugs within living organisms. Information on
structural formula, dissociation constant, water
solubility, hydrolysis, surface tension of the substance,
etc, can be obtained from partition coefficient.
It is useful in the study of the equilibrium
thermodynamics and is very useful to understand solutesolvent interactions in both aqueous and organic
solvents. Hydrophobic drugs with high partition
coefficients are preferentially distributed to hydrophobic
compartments (lipid bilayers of cells) while hydrophilic
drugs with low partition coefficients are preferentially
found in hydrophilic compartments (blood serum). Most
studies are concentrated on the water because water is
the major and most important solvent in biological
systems. The partition coefficient of a solute between
octanol and water is widely used to predict drug
pharmacokinetic properties2. Various techniques used to
measure the partition coefficients include the classical
shake flask method and stir flask methods, dual-phase
potentiometric titrations, reversed phase planer and
liquid chromatographic procedures, cyclic voltammeter,
centrifugal partition chromatography, counter-current
distribution, rotating diffusion cells, etc. The most
classical and reliable method of determination of
partition coefficient is the shake flask method, which
consists of dissolving some of the solute in question in

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equal volumes of aqueous and organic solvents and then

measuring the concentration of the solute in each solvent
by spectroscopic method3.

Adsorption is a phenomenon by which the molecules of

gas, vapor and liquid spontaneously concentrate at a
contacting surface without undergoing chemical
reaction, thereby it forms the surface layer. Adsorption
is one of the most important mechanisms of interaction
between drugs and excipients. The forces involved in
this type of interaction can be either physical or
chemical in nature or combination of both. When the
drug is in contact with adsorptive it can be adsorbed on
it causing reduction in their systemic absorption and
toxicity of drug4. Activated charcoal is commercially
available adsorptive and activated by heating. The
process of activating results in particles with small pores
having a large surface area (typically 950-2000 m2/g),
and are coated with carbon moieties that have varying
ability to form complex with drug molecules. The invitro adsorption of several drugs onto various
adsorptives has been studied with a variety of
techniques5.
Fig. 1: Freundlich adsorption isotherm of Fluconazole on
activated charcoal

Kim D. applied statistical adsorption isotherms for the

multilayer adsorption of gas molecules on non-porous
solid adsorbents.7 Robert et al. studied the adsorption of
tramadol hydrochloride on activated charcoal in-vitro
and in-vivo and found that 13 to 14 fold rightward
antinociceptive dose-response curve5. Sanjuan et al
studied the in-vitro adsorption of methotrexate and
calcium leucovorin onto cholestyramine and concluded
that it is possible to use cholestyramine by the oral route
in order to prevent reabsorption of methotrexate which is
excreted via bile, thus improving the elimination of drug
in the faeces8. Oremusova et al. studied the adsorption,
partition and release study of terbinafine hydrochloride
and claimed that adsorption follows the both the
adsorption isotherms.9 Teiko et al. studied the adsorption
of drugs on the various adsorbents and found that
medicinal carbon showed an excellent adsorption of all
the tested drugs.10 Otero et al. studied the effect of

temperature on adsorption process of salicylic acid on

polymeric adsorbents and activated charcoal11. Suhair et
al studied the adsorption of ketotifen on some of the
pharmaceutical excipients and suggested that the
predominant mechanism of ketotifen adsorption is
physical and exothermic in nature12. Andrzej et al.
studied the application of adsorption method to the
chromatographic analysis of free drug concentration13.

The process of adsorption is affected by various factors,

viz. time, pH, and specific surface area, cross linking
time, surface free energy and nature of adsorbents used4.
The equations to describe the adsorptions theoretically
well are the Langmuir and Freundlich adsorption
isotherms6. Fluconazole is a highly selective inhibitor of
fungal cytochrome P-450 by sterol C-14 alphademethylation. Fluconazole a bis-triazole antifungal
agent used in the treatment of superficial and systemic
infection. Mammalian cell demethylation is much less
sensitive to Fluconazole inhibition. It exhibits in vitro
activity against Cryptococcus neoformans and Candida
spp. In common with other azole antifungal agents, most
fungi show a higher apparent sensitivity to Fluconazole
in vivo than in vitro. There have been reports of cases of
superinfection with Candida species other than C.
albicans, which are often inherently not susceptible to
oral formulation. In such cases requires alternative
antifungal therapy. The drug has slight solubility in
water (8mg/ml) and phosphate buffer (10 mg/ml),
soluble in organic solvents at room temperature. It has
melting point of 138-142 C and is widely available as
tablets and IV infusion. It is well absorbed following
oral administration having bioavailability of 90% or
more compared to intravenous administration14.
In the present work, the partition coefficient of
Fluconazole in different organic solvents and phosphate
buffer pH 7.4 has been determined by shake flask
method where as adsorption studies on activated
charcoal and talc were performed at different
temperatures to fit the adsorption isotherm models to
investigate the suitability of the drug for the topical
effects.

2. EXPERIMENTAL:

2.1. Materials:
Fluconazole was a generous gift from Cipla
Pharmaceutical Ltd. Mumbai. Dichloromethane,
dichloroethane, hexanol and octanol were purchased
from Spectrochem Pvt. Ltd. Mumbai. The Activated
charcoal and Talc were purchased from Loba Chemie,
Mumbai and all other ingredients were of analytical
grade.
2.2
Calibration curve of Fluconazole:
A calibration curves as mean of five sets were obtained
by UV-visible spectrophotometer (Jasco, Japan). An
accurately weighed 10 mg of Fluconazole was dissolved
in 100 ml of each solvent system as listed in Table 1.

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Asian J. Research Chem. 2(2): April.-June, 2009

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These solutions are further diluted to 5, 10, 15, 20, 25,

30, 35 and 40 g/ml and absorbance was measured at
max 260 nm.
Fig. 2: Langmuir adsorption isotherm of Fluconazole on
activated charcoal

2.4.1
Activation of adsorptive:
Adsorptives were passed through sieve No.180 and
evenly spreaded in a Petri-dish followed by heating in
hot air oven at 100 C for 24 hours to make the surface
active for adsorption. The dried adsorptives were
allowed to cool to room temperature and used for the
study immediately.

2.4.2
Adsorption on talc and charcoal:
In each of stoppered glass bottle accurately weighed 1 g
of adsorptive were transferred. An aqueous stock
solution of Fluconazole (1 mg/ml) was prepared by
dissolving small quantity of drug in methanol and the
final volume was adjusted with phosphate buffer pH 7.4.
A specified quantity of stock solution was transferred to
each bottle to have a concentration of 1, 1.5, 2, 2.5, 3,
3.5, and 4 mg/ 100ml, respectively. The bottles were
immediately placed in incubator maintained at specified
temperatures (i.e. 25, 35 and 45C) and intermittently
agitated during the period of 1 hour to attain adsorption
equilibrium.
Table 1: Calibration curve - linear
values
Solvent system
Phosphate buffer pH 7.4 at 25C
Phosphate buffer pH 7.4 at 35C
Phosphate buffer pH 7.4 at 45C
Dichloromethane
Dichloroethane
Hexanol
Octanol

regression analysis
r
0.999
0.999
0.997
0.994
0.997
0.997
0.998

Slope
43.56
42.55
45.41
45.69
40.68
36.95
31.96

2.3.
Partition coefficient study15:
The partition coefficient of Fluconazole was studied
using various solvent systems listed in Table 2 by shake
flask method. Fluconazole was added continuously to
non aqueous phase till saturated solution is obtained.
The solution was filtered through Whatman filter paper.
The filtrate was used as stock solution to prepare
different strength solutions of which aliquots of 10, 20,
30 and 40 ml were transferred separately to different
flasks already containing 50 ml of phosphate buffer and
the final volumes were made to 100 ml using respective
pure non aqueous solvent. These flasks were shaken on
orbital shaker at room temperature (25C) for 1 hour to
attain chemical equilibrium. The solution mixtures were
separated using separating funnel. At the end, aqueous
and non-aqueous portions were analyzed for
Fluconazole
concentration
using
UV-visible
spectrophotometer at max 260 nm. The reliability of the
determined values of partition coefficients were tested
by comparison of the means of triplicate determinations
with overall mean.
2.4.
Adsorption Study15:
The adsorption of Fluconazole was studied using
hydrophobic adsorptives, activated charcoal and talc.

2.4.3
UV spectrometric analysis:
After completion of 1 hour the content of bottles were
filtered through the membrane filter (100 ) and filtrates
were analyzed by UV spectrophotometer at max = 260
nm.
Table 2: Results of partition coefficient of Fluconazole
Solvent Systems
Partition
coefficient*
Dichloromethane/ phosphate buffer pH 7.4
1.080.01
Dichloroethane/ phosphate buffer pH 7.4
1.050.01
Hexanol/ phosphate buffer pH 7.4
1.030.02
Octanol/ phosphate buffer pH 7.4
1.010.01
(*Results are the mean of triplicate observations SD)

3. RESULTS AND DISCUSSION:

Adsorption, partition and releasing balances of drug are

important properties which have to be known in the
development of dosage form. The study of partition
balances is useful in understanding the behavior of drugs
in the organism. Investigations of temperature
dependences of drug adsorption on hydrophilic or
hydrophobic surfaces are often used to obtain
information about the adsorption and adhesive
mechanisms which are directly related to its
physicochemical properties that also help in predicting
their compatibility and suitability.
In this work, effect of length of carbon chain on partition
process of Fluconazole in o/w system was studied. The
effect of temperature on adsorption of Fluconazole on
activated charcoal and talc at different temperatures was
studied in order to find out which isotherm describe more
appropriate experimental adsorption process. Before
formulating the any formulation we have to study their
physicochemical characteristics for their compatibility
and suitability.

215

Table 3: Adsorption
charcoal)
Temperature
(C)
25
35
45

Asian J. Research Chem. 2(2): April.-June, 2009

,

parameters of Freundlich and Langmuir adsorption isotherm at different temperature (activated

Freundlich adsorption isotherm
n
K
r
8.32
0.0074
0.997
10.32
0.0065
0.997
13.65
0.0042
0.996

Langmuir adsorption isotherm

b
a
r
91849
61.56
0.998
97405
76.43
0.998
103200
87.46
0.995

Table 4: Adsorption parameters of Freundlich and Langmuir adsorption isotherm at different temperature (Talc)
Freundlich adsorption isotherm
Langmuir adsorption isotherm
Temperature
(C)
n
K
r
b
a
r
25
9.71
0.0045
0.996
113891
137.66
0.993
35
11.65
0.0033
0.997
120879
143.31
0.992
45
12.87
0.0028
0.996
143446
162.68
0.994

Calibration curve of Fluconazole:

Fluconazole absorbance spectrum has two maxima at
260 and 267 nm. Determination of calibration curve and
all measurements for adsorption and partition were
carried out at max 260 nm. From the plot of absorbance
at max = 260 nm Vs concentration in various organic
solvents and at temperatures of 25, 35, 45C in
phosphate buffer (pH 7.4) it shows linear regression and
obeys Beers law. The linear regression line values in
different solvents are given in Table 1.
Fig. 3: Freundlich adsorption isotherm of Fluconazole on
talc

applications in the assessment of its penetration and

distribution.
Adsorption:
The effect of temperature on Fluconazole adsorption
from non-aqueous buffered solutions on Activated
charcoal and Talc I. P. at temperatures 25C, 35C and
45C were studied. The objective of this study was to
investigate which adsorption isotherm is more
appropriate for the experimental adsorption process.
From the observed values amount of solute adsorbed per
gram of activated charcoal and talc at all temperatures
were calculated. The calculated data indicate that
Fluconazole adsorption was lower at high temperature
than that at low temperature. It was also found that
higher amounts of Fluconazole were adsorbed on
activated charcoal than that of talc at all temperatures.
The experimental adsorption isotherms were expressed
by the Freundlich (equation 1) and Langmuir model
(equation 2) of adsorption6.
log w/m = log K + 1/n log C

Partition coefficient:
The partition coefficient study is useful in understanding
of the behavior of drug in the organism. Parameters of
the calibration curve were used for determination of the
experimental partition coefficients in the o/w partition
solvent systems. All the obtained values of partition
coefficient were given in Table 2.
The experimental observations showed that as the length
of carbon chain of lipophilic solvents increases (octanol
> hexanol > dichloroethane > dichloromethane) the
partition coefficient values goes on decreasing. The
observed partition coefficient values are very close to 1
suggesting the easy permeation through biological
membrane of the microorganism. The partition
coefficient of drug between octanol and water is a
physicochemical properties with wide spread

. (1)

Where, w/m is observed values of the Fluconazole in mg

adsorbed per gram of adsorbent, C is concentration of
Fluconazole in mg per 100 ml remaining in solution
after equilibrium had been established, K is amount of
Fluconazole adsorbed per one gram adsorbent at an
equilibrium concentration dependent on character of
adsorbent and adsorbate, and n is Freundlich parameter
dependent on temperature and related to Fluconazole
affinity to the adsorbent.

. (2)

Where, a is related to the surface area of the adsorbates

as amount of maximum monolayer adsorption and b is
constant called adsorption coefficient related with
enthalpy of adsorption. The experimental data was fitted
to Freundlich isotherm model as shown in Fig. 1 and 3.
The calculated data was also fitted to Langmuir isotherm
model, Fig. 3 and 4.

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Asian J. Research Chem. 2(2): April.-June, 2009

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According to Freundlich isotherm the plot at different

temperatures for adsorption of Fluconazole on activated
charcoal and talc is linear and provided n and K values as
shown in Table 3 and 4. The value n increased with
temperature of solutions and plot of n vs. temperature was
not found linear. Plot of K vs. temperature was found to
be nearly linear and decreased with increasing
temperature. All Langmuir plots for different
temperatures gave a and b values as listed in Table 3 and
4. The values a and b increased with increasing
temperature of solution. This clearly suggests that
adsorption of Fluconazole on activated charcoal and talc
conveniently describes both models of adsorption.

Fig. 4: Langmuir adsorption isotherm of Fluconazole on

talc

4.

5.

6.

7.

8.
9.
10.
11.
12.
13.

14.
15.

CONCLUSION:

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review. Drug Delivery Technology. 2006; 31: 58-63.
Robert BR, et al. Determination of the adsorption of
tramadol hydrochloride by activated charcoal in vitro and
in vivo. J Pharmacol Toxicology methods. 2000; 43: 205210.
Hajare AA and Pishawikar SA. Comparative in vitro
adsorption studies of diclofenac sodium and dilitiazem
hydrochloride by talc. The Indian Pharmacist. 2006; 47: 8184.
Kim D. Statistically modeled adsorption isotherms for the
multilayer gas molecules adsorbed on non porous solid
adsorbents of two and three groups sites. Korean J Chem
Eng. 2000; 17: 156-168.
Sanjuan M, et al. Adsorption of methotrexate and calcium
leucovorin onto chlolestyramine in vitro. Int J Pharm.
2004; 278: 283-291.
Oremusova J and Vitkova Z. Adsorption, partition and
release balances of terbinafine hydrochloride, Part I.
Pharmazie. 2007; 62: 273-277.
Teiko T, et al. In vitro study of the adsorption
characteristics of drugs. Biol Pharm. Bull. 2005; 28: 841844.
Otero M, Carlos A and Alirio ER. Adsorption of salicylic
acid onto polymeric adsorbents and activated charcoal.
Reactive and Functional Polymers. 2004; 60: 203-213.
Suhair SA, et al. Adsorption of ketotifen onto some
pharmaceutical excipients. Int J Pharm. 1997; 149: 115121.
Andrzej LD, Mateusz K and Rafal K. Application of
adsorption method to the chromatographic analysis of free
drug concentration. J Pharm Biomed Analysis. 2006; 41:
973-978.
Susan B. Merck Index. Edn12. Whitehouse Station, NJ:
Merck and co. Inc 1996, 698.
More HN and Hajare AA. Practical Physical Pharmacy.
Career Publications. 2007.

Experimentally determined partition coefficients

increased with the length of carbon chain in the
partitioning system. Based on results obtained it could
be concluded that n-octanol /water system is the most
suitable than other organic solvent systems. This is
because the partition coefficient obtained in noctanol/water system is nearly equal to 1. The
Freundlich and Langmuir adsorption isotherms showed
the adsorption of Fluconazole on talc and activated
charcoal at different temperatures conveniently
describing both models. Partition and adsorption data
suggest that Fluconazole can be formulated as topical
dosage form for local effects for the increasing
therapeutic action.

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