UV Spectroscopy Analysis for Itraconazole
Taranjeet Kukreja1,
Swarnlata Saraf *
1University
Institute of Pharmacy, Pt. Ravi Shankar Shukla University, Raipur - 492010,
Chhattisgarh, India
Abstract:
Itraconazole
is a triazole antifungal agent that is synthesised. Itraconazole has been
manufactured into a variety of pharmacological formulations and administered in
a variety of ways. Itraconazole pills are used to treat pulmonary fungi that
can cause fungal infection and spread throughout the body. Because Itraconazole
is not yet officially listed in any pharmacopoeia, only a few procedures for
quality control and stability testing in pharmaceutical formulations have been
published. The goal of this study is to develop a more precise, easy, and
cost-effective spectrophotometric approach for analysing Itraconazole in bulk
and capsule dosage forms with improved precision, accuracy, and sensitivity.
The UV spectroscopic determination was performed with Chloroform as the solvent
at an absorption maximum of 267 nm. Linearity over the concentration range in
the UV spectroscopic approach. The linearity of Itraconazole over the
concentration range was found to be 1-10 g/ml using the UV spectroscopic
technique, with a correlation coefficient of 0.999. The findings of the
analyses were statistically and the recovery studies have confirmed this.
Keywords:
Itraconazole, pharmacopoeia, UV spectroscopic technique, Quality control.
Introduction
Itraconazole
is a triazole antifungal agent that is synthesised. Itraconazole is a racemic
combination of four diastereomers (two enantiomeric pairs), each with three
chiral centres, in a ratio of 1:1:1:1. The following nomenclature can be used
to denote it: 4-[4-[4-[4-[[ 2. (2, 4-dichlorophenyl) - two (1H-1, 2, 4- triazole-
1-ylmethyl) - 1,3- dioxolan-4-yl] methoxy]phenyl] piperazine-1- yl]phenyl] piperazine-1-
yl]phenyl] piperazine-1- yl]phenyl] piperazine-1- yl]phenyl] piperazine-1-
-2-(1-methyl propyl) -2, 4-dihydro-1, 2-dihydro-1, 2-dihydro-1, 2-dihydro-1,
2-dihydro-1, 2-dihydro (Fig 1).
Figure1:
Itraconazole's structure
C35H38Cl2N8O4
is the molecular formula and the chemical molecular weight is noted to be705.64.
[1-4] It's a powder that's white to slightly yellowish. In alcohols, it is very
partially soluble, but in dichloromethane, it is completely soluble.
Itraconazole is an extremely lipophilic compound that is water
insoluble. It's a very weak base (pKa =3.7) that only ionises at very low pH
levels. It's a three-chiral hydrophobic antimycotic medication that's employed
in clinical trials as a stereoisomeric combination. [5] It is an oral triazole
antifungal medication efficacious against dermatophytes, species of bacteria,
Malassezia, and penicillium species having a broad range of activity and
Histoplasma capsulatum var. capsulatum, among other fungal species. [6,7]
Mechanism
of action of Itraconazole
It
binds to 14-demethylase, a cytochrome P-450 enzyme that is required for the
conversion of lanosterol to ergosterol. Because ergosterol is a necessary
component of the fungal cell membrane, inhibiting its synthesis increases
cellular permeability, causing cellular contents to flow out. Itraconazole can
also decrease endogenous respiration, interact with membrane phospholipids, prevent
yeasts from transforming into mycelial forms, block purine uptake, and disrupt
triglyceride and/or phospholipid production. When itraconazole is taken with
food, its oral bioavailability increases and plasma concentrations are nearly
twice as high as when taken when fasting. Itraconazole is metabolised primarily
in the liver by the cytochrome p450 3a4 isoenzyme system, which produces
various metabolites, the most significant of which is hydroxyl itraconazole 8. [9]
It is metabolised in the liver, primarily through an oxidative pathway,
forming the bioactive metabolite hydroxyl itraconazole. [10]
Materials
and methods
Experimental
Chemicals
and reagents: Throughout UV spectrophotometric technique, development and
validation Chloroform was used.
Instrumentation
UV
spectrophotometric technique was performed on a double beam UV-visible
spectrophotometer (Shimadzu, model 1800) having two matched quartz cells with a
1 cm light path.
Selection
of solvent
For
the analysis of Itraconazole, Chloroform had been selected as the ideal solvent
for spectrophotometry.
Standard
stock solutions preparation
A 10 mg
Itraconazole reference standard was accurately weighed and transferred to a 10
ml volumetric flask, where it was dissolved and diluted up to the mark using
chloroform to yield a stock solution with a strength of 500g/ml. Diluting 1 ml
of stock solution to 5 ml with chloroform yielded a 50 g/ml working standard
solution.
Preparation
of Sample stock solution
For
analysis of the drug, 10mg of the drug itraconazole was weighed and transferred
to a 10ml volumetric flask and dissolved with chloroform. The itraconazole drug
solution was diluted to get a final concentration of 10μg/ml. The absorbance of
these solutions was measured at 267 nm. The amount of Itraconazole was
calculated using the calibration curve.
Formula:
%Purity=Sample absorbance / Standard
absorbance X 100
1. Method
validation
The
method was validated according to the International Conference on Harmonization
(ICH)
Q2B
guidelines 1996 for validation of analytical procedure to determine the
linearity,
limit
of detection, accuracy and precision.
2. Linearity
& Range
Under
the experimental conditions, the calibration graphs of the absorbance versus
Concentration
was found to be linear over the range of 0.2-1.0μg/ml for the proposed method.
The
statistical analysis of data obtained for estimation of Itraconazole is
indicated chloroform
of
accuracy for the proposed methods evidenced by the low values of standard
deviation and
coefficient
of variation. The results are noted below:
Table
1: Itraconazole Linearity Data
S.
No.
|
Concentration
in μg/ml
|
Absorbance
in UV
|
1
|
0
|
0
|
2
|
2
|
0.1316
|
3
|
4
|
0.2520
|
4
|
6
|
0.3527
|
5
|
8
|
0.4713
|
6
|
10
|
0.5813
|
|
y = 0.0575x + 0.0106
slope = 0.057
RSQ = 0.9986
|
|
Results
and Discussion
The
purpose of this study was to validate an Itraconazole using a
UV-Spectrophotometric technique under optimal conditions. The validation
parameters' results were found to be within acceptable limits. Within the
concentration range of 1-10g/ml, itraconazole followed linearity. The measured
linearity range suited Beer-law Lambert's well, and the corresponding
regression coefficient (r=0.999) indicates a high degree of technique
sensitivity, as shown in Table 1. The number of drugs detected and the findings
of the analysis demonstrate that the percentage of the drugs found and the number
of drugs found was in good accord.
Conclusion
UV-Spectrophotometer
techniques produced equivalent results. The linearity of Itraconazole over the
concentration range was found to be 1-10 g/ml using the UV spectroscopic
technique, with a correlation coefficient of 0.999. The findings of the
analyses were statistically and the recovery studies have been confirmed.
Acknowledgment
I am thankful to
the University Institute of Pharmacy (UIOP), Pt. Ravishankar Shukla University,
Raipur, Chhattisgarh, India for my practical support. I also want to thank
Prof. Swarnlata Saraf, University Institute of Pharmacy (UIOP), Pt. Ravishankar
Shukla University, Raipur, Chhattisgarh, India for her valuable suggestions and
guidance.
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