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ORIGINAL ARTICLE
a,*
Department of Chemistry, Chodhary Dilip Singh Kanya Mahavidyalya, Bhind 477001, MP, India
School of Pharmacy, Devi Ahilya Vishwavidyalaya, Indore 452001, MP, India
Department of Pharmaceutical Sciences, Dr. H.S. Gour University, Sagar 470 003, MP, India
KEYWORDS
Nitazoxanide;
Ooxacin;
HPTLC;
Spectrophotometric method;
ICH
* Corresponding author.
E-mail address: drsmita.sharma@rediffmail.com (S. Sharma).
Peer review under responsibility of King Saud University.
1. Introduction
Ooxacin (OFL) is chemically 9-uoro-2,3-dihydro-3-methyl10-(4-methyl-1-piperazinyl)-7-Oxo-7H-pyrido (1,2,3-di)-1,4benzoxazine carboxylic acid. It is a uoroquinolone derivative.
It is used mainly as an antibacterial. It is ofcial in the United
1878-5352 2012 King Saud University. Production and hosting by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.arabjc.2012.07.009
Please cite this article in press as: Sharma, S. et al., Simultaneous determination of Nitazoxanide and Ooxacin in pharmaceutical preparations
using UV-spectrophotometric and high performance thin layer chromatography methods. Arabian Journal of Chemistry (2012), http://
dx.doi.org/10.1016/j.arabjc.2012.07.009
2
State Pharmacopoeia, 2006. Literature survey reveals that
spectrophotometric, HPLC, RP-HPLC, and HPTLC (Rane
et al., 2008; Kasture et al., 2004; Gopu et al., 2007; Kraas
and Hirrle, 1986; Gandhimathi et al., 2006) methods are available for the determination of Ooxacin from pharmaceutical
preparations and biological formulation. Nitazoxanide (NT),
N-(5-nitro-2-thiazolyl) salicylamide acetate (ONeil, 2001;
Reynolds, 2002) is a Nitrothiazole derivative. Its chemical
structure is related to metronidazole. It is a broad spectrum
antiprotozoal. Nitazoxanide is an antiamebic and anthelmintic
agent. It is indicated for amebiasis, helminthiasis, giardiasis,
fascioliasis, trichomoniasis and cryptosporidiosis, including
those with AIDS or HIV infections (Yoshimasa et al., 1996;
Cavier, 1978). Literature survey reveals that, spectrophotometric and RP-HPLC (Kapse et al., 2006; Narayana et al., 2006)
methods are available for the estimation of Nitazoxanide in
single dosage form. A survey of the literature reveals of these
combination that a variety of spectrophotometric and chromatographic methods (Singh et al., 2011; Game and Sakarkar,
2011; Lalitha et al., 2009; Mahaparale et al., 2009). An Ooxacin and Nitazoxanide combination is indicated to have
antibacterial and antiprotozoal activities. The combination
of Nitazoxanide and Ooxacin is antiparasitic and antibacterial which is effective against a wide variety of protozoa, helminthes and Gram-negative organisms. Oral bioavailability
is good and well tolerated, with mild gastrointestinal side effects. Used in Giardia intestinal is induced diarrhea in patients
(Guerrant et al., 2005). A combination of 200 mg of ooxacin
and 500 mg of Nitazoxanide is available commercially as
tablets (Nitazete-O). The aim of this paper was to explore
the possibility of techniques of simultaneous estimation using
UV spectrophotometric, rst derivative and HPTLC methods
for quantifying Ooxacin and Nitazoxanide simultaneously in
their mixture forms. The proposed methods are simple, convenient, precise, accurate, and economical than the reported
method. All chemicals and reagents used are of analytical
grade and were purchased from Merck Chemicals, India. All
dilutions were performed in standard volumetric asks. Pure
and tablet dosage form, Nitazete-O (claim: 500 mg NT and
200 mgOF) was procured from the local market (see. Fig. 1).
2. Experimental
2.1. Instrumentation and chromatographic conditions
Chromatography was performed on 10 cm 10 cm precoated
silica gel 60 F254 HPTLC plates. The chromatographic plates
were prewashed with methanol and dried in an oven at
120 C for 2 h before use. The samples were applied onto the
plates as a band with 6 mm width using Camag 100 ll sample
syringe (Hamilton, Switzerland) with a Linomat 5 applicator
(Camag, Switzerland). Linear ascending development was carried out in a twin trough glass chamber (for 10 10 cm). Densitometric scanning was performed using Camag TLC scanner
3 in the range of 4001500 ng/spot and operated by winCATS
software (V 1.4.2, Camag). The chromatography estimation
was performed using the following conditions: stationary
phase was precoated with silica gel 60 F254 aluminum sheets
and the mobile phase used was chloroform:carbon tetra chloride:toluene:glacial acetic acid (10:5:3:0.5 v/v). The source of
radiation was a deuterium lamp. Slit dimensions were
S. Sharma et al.
Figure 1
Please cite this article in press as: Sharma, S. et al., Simultaneous determination of Nitazoxanide and Ooxacin in pharmaceutical preparations
using UV-spectrophotometric and high performance thin layer chromatography methods. Arabian Journal of Chemistry (2012), http://
dx.doi.org/10.1016/j.arabjc.2012.07.009
Table 1
Parameter
Repeatability
Precision (a) Intra-day
(b) Inter-day
RF (SD)
Linearity and range (ng/spot)
Linearity detection (ng/spot)
Limit of quantication (ng/spot)
% Accuracy SDa (n = 6) (%)
LOD
LOQ
UV method-I
UV method-II
HPTLC
NTO
OFL
NTO
OFL
NTO
OFL
0.874
2.65
1.64
101.04
0.021
0.012
0.577
1.63
0.48
99.96
0.065
0.041
0.681
0.954
1.21
99.98
0.144
0.095
0.317
0.215
0.62
100.08
0.165
0.0.81
0.451
1.45
0.154
0.57
4002500
112
164
99.94
5.3
26.00
0.654
0.974
0.654
0.63
4001200
150
217
99.85
6.5
12.32
Please cite this article in press as: Sharma, S. et al., Simultaneous determination of Nitazoxanide and Ooxacin in pharmaceutical preparations
using UV-spectrophotometric and high performance thin layer chromatography methods. Arabian Journal of Chemistry (2012), http://
dx.doi.org/10.1016/j.arabjc.2012.07.009
S. Sharma et al.
Table 2
Method
Tablet content
UV-I
NTO
OFL
NTO
OFL
NTO
OFL
UV-II
HPTLC
500
200
500
200
500
200
Amount Founda
(in mg)
(In%)
499.974
199.053
500.03
199.31
499.941
200.31
99.994
99.931
100.05
99.986
99.941
100.31
SDa
RSD (%)a
0.041
0.035
0.143
0.087
0.198
0.142
0.18
0.051
0.217
0.328
0.054
0.083
Table 3
80
100
120
Drug
NTO
OFL
NTO
OFL
NTO
OFL
HPTLC
UV-Method I
UV-Method II
Recovery (%)*
SD*
Recovery (%)*
SD*
Recovery (%)*
SD*
100.36
100.27
100.01
99.93
99.97
100.12
0.043
0.154
0.049
0.0654
0.044
0.219
99.95
99.97
100.04
100.06
100.08
99.94
0.033
0.0761
0.0207
0.0381
0.0549
0.0087
100.07
100.21
99.96
100.09
99.90
99.82
0.0321
0.0543
0.0612
0.0431
0.0543
0.0451
simultaneous equation method, wavelengths selected for quantitation were 221.8 nm (kmax of Nitazoxanide) and 244.3 nm
(kmax of Ooxacin). In rst order derivative spectroscopy,
wavelengths selected for quantitation were 263.6 nm for Nitazoxanide (zero cross for Ooxacin) and 269.2 nm for Ooxacin
(zero cross for Nitazoxanide). For UV spectrophotometric
method, linearity was obtained in the concentration range of
525 lg/ml, for both drugs; with regression 0.9994 and
0.9997, intercept 0.0854 and 0.0328 and slope 0.0188 and
0.0742 for Nitazoxanide and Ooxacin, respectively. For UV
Spectrophotometric method LOD for Nitazoxanide and
Ooxacin was found to be 0.021 and 0.065 lg/ml respectively.
LOQ for Nitazoxanide and Ooxacin was found to be 0.012
and 0.041 lg/ml respectively. These data show that microgram
quantity of both drugs can be accurately determined. Robustness of the method when small changes in the mobile phase
composition (0.1 ml for each component) were made and
the effects on the results were examined.
4. Conclusion
The proposed HPTLC method was validated as per ICH
guidelines. The standard deviation, %RSD and standard error
calculated for the method are low, indicating a high degree of
precision of the methods. The results of the recovery studies
performed show a high degree of accuracy of the proposed
methods. The proposed method is highly accurate, selective
and precise hence can be used for a routine quality-control
analysis and quantitative simultaneous determination of
Nitazoxanide and Ooxacin in pharmaceutical preparations.
Moreover, the proposed method has the advantages of simplicity, convenience and quantication of Nitazoxanide and
Ooxacin in combination and can be used for the assay of their
dosage form.
Acknowledgements
We are grateful to Prof. D.V. Kohli and Prof. Abhay Kumar
Singhai Department of Pharmaceutical Sciences Dr. Harisingh
Gour Sagar University Sagar (M.P.) India, for given valuable
suggestion and facility.
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Please cite this article in press as: Sharma, S. et al., Simultaneous determination of Nitazoxanide and Ooxacin in pharmaceutical preparations
using UV-spectrophotometric and high performance thin layer chromatography methods. Arabian Journal of Chemistry (2012), http://
dx.doi.org/10.1016/j.arabjc.2012.07.009
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Please cite this article in press as: Sharma, S. et al., Simultaneous determination of Nitazoxanide and Ooxacin in pharmaceutical preparations
using UV-spectrophotometric and high performance thin layer chromatography methods. Arabian Journal of Chemistry (2012), http://
dx.doi.org/10.1016/j.arabjc.2012.07.009