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International Journal of ChemTech Research
CODEN( USA): IJCRGG
ISSN : 0974-4290
Vol.2, No.4, pp 1939-1944,
Oct-Dec 2010

Development and Validation of Stability indicating
RP-HPLC Method for the estimation of Azithromycin
Suspension
Sachin Kumar Singh*, Dev Prakash, Tamizh Mani T
Bharathi College of Pharmacy, Bharathinagara, Maddur Tq, Mandya Dist.,
Karnataka- 571422, India
*Corres.author:singhsachin23@gmail.com
Phone No. 07259653939

Abstract: A RP-HPLC method was developed and validated for quantitative determination of azithromycin in

pharmaceutical suspension dosage forms. The chromatography was carried out on a Phenomenex C (150 x 4.6 mm i.d.,
18

5μ) column with Acetonitrile: 0.5 % Formic acid as mobile phase (Isocratic A: B = 40: 60 % v/v), at 215 nm detector
wave length with aflow rate of 1 ml/min. Clarithromycin was used as an internal standard. The linearity was established
in the range of 20 - 600 ng/ml for HPLC. The HPLC method was accurate and precised for azithromycin suspension
with a recovery of 98.75 to 99.44%. The spiked sample solutions were stable upto 1 month. This validated method can
be used for estimation of azithromycin in pharmaceutical suspension.
Keywords: Azithromycin, Reversed Phase High Performance Liquid Chromatography, Analytical method validation,
Pharmaceutical solid dosage forms.

Introduction

Azithromycin1 is chemically (2R, 3S, 4R, 5R, 8R,
10R,11R,12S,13S,14R)-13-[(2,6-dideoxy -3 - C methyl - 3 - O - methyl - α- L - ribo - hexopyranosyl)
oxy]-2-ethyl-3, 4, 10-trihydroxy-3, 5, 6, 8, 10, 12, 14heptamethyl - 11 - [[3, 4,6-trideoxy-3(dimethylamino)-β-D-xylo-hexopyranosyl]oxy]-1-oxa6-azacyclopentadecan-15-one. It is derived from
erythromycin; however, it differs chemically from
erythromycin in that a methyl-substituted nitrogen
atom is incorporated into the lactone ring.
Azithromycin, like all macrolyte antibiotics, prevent
bacteria from going by interfering with their ability to
make protein. Due to the differences in theway
proteins are made in bacteria and humans, the
macrolyte antibiotics do not interfere with humans’
ability to make proteins. It binds to the 50s rRNA
subunit of the 70s bacterial ribosome’s, therefore
inhibits RNA-dependent protein synthesis. It inhibits

the translation of mRNA in bacterial cells at the chain
elongation step; result in the blockage of
transpeptidation. Nucleic acid synthesis isnot affected.
Various analytical methods2-19 like UV, HPLC,
HPTLC and LC/MS have been reported for the
determination of azithromycin in its tablet or capsule
dosage forms either in single or in combination.
There is however no reported HPLC method for the
analysis of azithromycin in suspension dosage forms.
This paper describes a validated HPLC method for the
quantitative determination ofazithromycin suspension
in reverse phase mode using Clarithromycin as an
internal standard. The proposed HPLC method
fulfilled the requirements of analytical parameters
necessary to be applied to the content uniformity tests
for finished pharmaceutical products in the study and
hence can be successfully applied for routine quality
control.

Sachin Kumar Singh et al /Int.J. ChemTech Res.2010,2(4)

Figure1: Structure of Azithromycin
Clarithromycin (Internal Standard)

and

1940

rate at 215 nm using UV detector. Spinchrom software
was used for the data interpretation.
Preparation of Standard Solutions
The different concentrations of standard solutions were
prepared to contain 20.0 to 600.0 ng/mL of
Azithromycin containing 40.00 μg/mL of internal
standard. These solutions were analysed and the peakareas and response factors were calculated. The
calibration curve was plotted using response factor Vs
concentration of the standard solutions. All solutions
were filtered through 0.45 μ membrane filter prior to
use.
Preparation of the Sample Solutions
The contents of azithromycin suspension were taken.
The Suspension containing equivalent to 25 mg of
azithromycin was accurately weighed and...