Grewal et al., IJPSR, 2012; Vol. 3(8): 2716-2720
IJPSR (2012), Vol. 3, Issue 08
ISSN: 0975-8232
(Research Article)
Received on 24 April, 2012; received in revised form 14 May, 2012; accepted 21 July, 2012
SIMULTANEOUS SPECTROPHOTOMETRIC ESTIMATION OF CIPROFLOXACIN AND ORNIDAZOLE IN TABLET
DOSAGE FORM
A.S. Grewal*1, S.K. Patro 2, S.K. Kanungo 2 and S.K. Bhardwaj 1
JCDM College of Pharmacy 1, Sirsa- 125 055, Haryana, India
Institute of Pharmacy & Technology 2, Salipur- 754 202, Cuttack, Orissa, India
ABSTRACT
Keywords:
Ciprofloxacin,
Ornidazole,
Spectrophotometry,
Simultaneous,
Validation
Correspondence to Author:
A.S. Grewal
Assistant Professor, JCDM College of
Pharmacy, Sirsa- 125 055, Haryana, India
Email: ajmergrewal2007@gmail.com
Three simple sensitive, rapid, accurate and economical methods were
developed for simultaneous estimation of Ciprofloxacin & Ornidazole in bulk
and its pharmaceutical formulations. First method was based on the
simultaneous equation, second method was based on Q-Analysis
(Absorbance ratio method) and third method was based on multicomponent
mode. The absorption maxima for Ciprofloxacin and Ornidazole were 272 nm
and 319 nm in 0.1 N NaOH respectively. The iso-absorptive point for
Ciprofloxacin Hydrochloride and Ornidazole was 292 nm. The calibration
curve was linear over the concentration range 2.5 to 22.5 µg/ml of
Ciprofloxacin & Ornidazole. The methods had been validated statistically and
by recovery study. It can be used in the simultaneous estimation of
Ciprofloxacin and Ornidazole in bulk drugs and its pharmaceutical
preparations in a routine manner.
INTRODUCTION:
Ciprofloxacin,
chemically,
1cyclopropyl-6-fluoro-1, 4- dihydro-4-oxo- 7 (piperazin1-yl) quinoline-3-carboxylic acid1; belongs to the group
of synthetic fluoroquinolone antibiotics with broad
antimicrobial activity. Literature review reveals
spectrophotometry 1-11, titrimetry 12, HPLC 13-18 and
HPTLC 19 for its determination. Ornidazole, chemically,
1-chloro-3-(2-methyl-5-nitro-1H-imidazol-1-yl) propan2-ol, is an antimicrobial agent.
Ornidazole is used in the treatment of amoebiasis and
other protozoal diseases. This drug is under the
category antihelmentics. The combination of two drugs
is not official in any pharmacopoeia; hence no official
method is available for the estimation of ciprofloxacin
and ornidazole in their combined dosage form.
Literature review reveals that few methods are
available for the determination of ornidazole in bulk
and solid dosage form based on spectrophotometry 2027
and RP-HPLC.
Ciprofloxacin and ornidazole in combined tablet
dosage form is available in the market, has gained
increasing acceptance in bacterial and protozoal
infections. Spectrophotometric 28-29 and HPLC 30 method
have been established for their simultaneous
estimation in tablet dosage form. This paper presents
three simple, sensitive, accurate and reproducible
spectrophotometric methods for simultaneous
determination of ciprofloxacin and ornidazole in tablet
dosage form.
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CIPROFLOXACIN
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Grewal et al., IJPSR, 2012; Vol. 3(8): 2716-2720
ORNIDAZOLE
FIGURE 1: STRUCTURE OF CIPROFLOXACIN AND ORNIDAZOLE
MATERIALS AND METHODS:
ISSN: 0975-8232
concentration range of 2.5 to 22.5 µg/ml were
prepared in 0.1 N NaOH using working standard
solution. The absorbance of resulting solution was
measured at 278 nm and 319 nm for both
Ciprofloxacin HCl and Ornidazole, and calibration
curves plotted at these wavelengths. Both the
drugs obeyed linearity individually within the
concentration range 2.5 - 22.5µg/ml. The overlay
UV spectrum of Ciprofloxacin Hydrochloride and
Ornidazole is shown in Fig. 2.
Instrument used: A Shimadzu UV-Vis Double beam
spectrophotometer (Pharmaspec-1700) with matched
quartz cells was used for all spectral measurements.
Materials: The tablets of the said combination were
purchased from a local pharmacy (The label claim
contained 500 mg of Ciprofloxacin and 500 mg of
Ornidazole). Sodium hydroxide was of analytical grade
chemical and distilled water was used to prepare all
solutions.
Preparation of 0.1N Sodium Hydroxide Solution: 0.1 N
NaOH was prepared by dissolving 4 g of sodium
hydroxide in sufficient distilled water to produce 1000
ml.
Preparation of Standard Solutions: The standard stock
solution of Ciprofloxacin and Ornidazole were
prepared by dissolving 50 mg of each drug in 50 ml of
0.1 N NaOH. Stock solutions of Ciprofloxacin and
Ornidazole were further diluted in 0.1 N NaOH to get
working standard solutions of concentration 100
µg/ml.
Method-I: Si ulta eous e uatio
ethod Vie odt’s
method): If a sample contains two absorbing drugs (X
and Y) each of which absorbs at the Imax of the other, it
may be possible to determine both drugs by the
technique of simultaneous equation Vie odt’s
method) provided that certain criteria apply.
a) Study of Beer’s-Lamberts law: The standard
solutions of Ciprofloxacin Hydrochloride (20 µg/ml)
and Ornidazole (20 µg/ml) in 0.1 N NaOH were
s a ed i the e ti e UV a ge to dete i e max
of oth the d ugs. The max of Ciprofloxacin
hydrochloride and Ornidazole were found to be
272 nm and 319 nm, respectively. A series of
standard solutions were prepared having
FIGURE 2: THE OVERLAY UV SPECTRUM OF CIPROFLOXACIN
HYDROCHLORIDE AND ORNIDAZOLE
b) Estimation of Absorptivity (E 1%, 1cm) values at
selected wavelengths: The Absorptivity (E 1%, 1cm
value) of Ciprofloxacin HCl and Ornidazole drugs
were calculated at 272 nm and 319 nm. Two
simultaneous equations were formed using these
values as given below.
CX = 0.0145714 A1 – 0.0050635 A2 …………..
CY = 0.039518 A2 – 0.0145311 A1 …………....
Where, A1 and A2 are the absorbance of sample at 272
nm and 319 nm respectively. CX and CY are the
concentration of Ciprofloxacin HCl and Ornidazole in
sample respectively.
c) Analysis of standard mixture: The mixture of pure
Ciprofloxacin HCl (10 µg/mL) and Ornidazole (10
µg/mL) was prepared from stock solution in 0.1 N
NaOH. Their absorbance value at the two selected
wavelength was recorded and quantitative
estimation of the drugs was carried out by solving
simultaneous equations.
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Grewal et al., IJPSR, 2012; Vol. 3(8): 2716-2720
d) Analysis of Tablet Formulation: Twenty tablets
were weighed and finely powdered. An accurately
weighed quantity of the powder equivalent to
about 50 mg of Ciprofloxacin & Ornidazole; was
taken in 50 mL volumetric flask and dissolved in 25
mL of 0.1 N NaOH ; it was further diluted up to the
mark with same solvent. The solutions were then
filtered and filtrate gets diluted to get final 10
µg/mL concentration of Ciprofloxacin HCl &
Ornidazole. The solution was then read at two
selected wavelength. The results of tablet analysis
are given in Table 1.
Method-II: Q - Analysis (Absorbance Ratio Method):
Q-Absorbance method depends on the property that,
for a substance which obeys Beer's law at all
wavelength, the ratio of absorbances at any two
wavelengths is a constant value independent of
concentration or path length. In the quantitative assay
of two components in a mixture by the absorbance
ratio method, absorbances are measured at two
a ele gths: O e ei g the max of one of the
o po e t 2) and the other being a wavelength of
equal absorptivities of the two components i.e. an isoabsorptive point.
a) Determination of Iso-absorptive point and
selection of suitable Wavelength: An isoabsorptive point (a wavelength of equal
absorptivity of the two components) was
determined by taking overlain spectrum of the
solutions ciprofloxacin HCl and Ornidazole (20
µg/ml each) in 0.1 N NaOH in UV range against the
solvent blank. From the overlain spectra of the two
d ugs, it as fou d that ip oflo a i sho ed max
at 7
a d o idazole sho ed max at 319 nm.
Iso-absorptive point was found out at 292 nm. The
two wavelengths 272 nm at which ciprofloxacin
exhibits maximum absorption and 292 nm as isoabsorptive point were selected for estimation of
drugs simultaneously.
b) Study of Beer-Lambert’s law: The solutions having
concentrations in range 2.5 - 22.5 µg/ml for both
ciprofloxacin HCl and ornidazole were prepared in
0.1 N NaOH using working standard solution. The
absorbances of resulting solutions were measured
at 272 nm and 292 nm. Calibration curves were
plotted at these wavelengths. Both the drugs
ISSN: 0975-8232
obeyed linearity individually and combination
within the concentration range of 2.5 - 22.5 µg/ml
for both ciprofloxacin & Ornidazole.
c) Estimation of Absorptivity (E 1%, 1cm) values at
Selected Wavelengths: The Absorptivity (E 1%,
1cm value) of Ciprofloxacin HCl and Ornidazole
drugs were calculated at 272 nm and 319 nm.
d) Analysis of Standard Mixture: The laboratory
mixture of Ofloxacin (5 µg/mL) and ornidazole (12
µg/mL) was prepared from stock solution in 0.5 N
acetic acid The absorbance value at the two
selected wavelength was recorded The contents of
Ciprofloxacin and Ornidazole were calculated by
substituting values in the formulae given below.
Cx = (QM - QY) A1 / (QX - QY) ax1
Cy = (QM – Qx) A1 / (Qy – Qx) ax1
Where,
Cx=concentration
of
Ciprofloxacin;
Cy=concentration of Ornidazole; Qx= ratio of
absorptivity of ciprofloxacin at 292 nm and 272 nm;
Qy= ratio of absorptivity of Ornidazole at 292 nm and
272; QM= absorbance ratio of mixture at 292 nm and
272 nm; ax1 and ay1=absorptivity of pure Ciprofloxacin
and Ornidazole respectively at iso-absorptive
wavelength.; A1=absorbance of mixture at isoabsorptive point.
e) Analysis of tablet formulation: Twenty tablets
were weighed and finely powdered. An accurately
weighed quantity of the powder equivalent to
about 50 mg of ciprofloxacin was taken in 50 ml
volumetric flask and dissolved in 25 ml of 0.1 N
NaOH; it was diluted up to the mark with 0.1 N
NaOH. The solutions were then filtered and filtrate
gets diluted to get final concentration of
ciprofloxacin HCl (10 µg/ml) and Ornidazole (10
µg/ml). The solution was then read at two selected
wavelength. The results of tablet analysis are given
in Table 1.
Method-III: Multi-component mode: From the
overlain spectra of the two drugs, the two wavelengths
selected as sampling wavelengths were 272 and 319
nm which are absorbance maxima for Ciprofloxacin
HCL and Ornidazole respectively. Now the sampling
wavelengths and concentrations of the two
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Grewal et al., IJPSR, 2012; Vol. 3(8): 2716-2720
components in each of the mixed standards were fed
to the instrument using multi-component mode of the
instrument and all the seven mixed standards were
scanned in the range of 400 to 200nm. The instrument
collects and compiles the spectral data from the mixed
standards and gets ready for the quantitative analysis
of the samples.
Validation of Analytical Methods: As per I.C.H.
guidelines.
Linearity and range: 2.5 – 22.5 µg/ml
Precision: For evaluation of precision of the results a
o e t atio of
g/ l as e aluated
4 epli ate
determinations. S.D. was found to be less than 0.1.
Precision study data is given in Table 2.
Accuracy (Recovery study): It was found out by
recovery study using standard addition method.
Known amounts of standard Ciprofloxacin and
Ornidazole was added to pre-analyzed samples at a
level from 80% up to 120% and then subjected to the
proposed methods. Results of recovery studies are
shown in Table 3.
TABLE 1: ANALYSIS OF CIPROFLOXACIN AND ORNIDAZOLE IN TABLETS
Label claim (mg/tab)
Amount found* (mg/tab)
Method
CIP
ORD
CIP
ORD
I
500
500
502.87
502.48
II
500
500
502.87
501.45
III
500
500
502.52
502.57
*Average of five determinations, CIP-Ciprofloxacin, ORD-Ornidazole
TABLE 2: PRECISION DATA OF CIPROFLOXACIN AND ORNIDAZOLE
Drug used
Conc. (µg/ml)
Ciprofloxacin (272 nm)
Ornidazole (319 nm)
ISSN: 0975-8232
% Label claim* ± S.D.
CIP
ORD
100.57±0.977
100.49±0.255
100.57±0.381
100.50±0.693
100.51±0.86
100.52±0.198
Absorbance
10
0.763
10
0.774
10
0.758
10
0.768
10
0.318
10
0.334
10
0.328
10
0.321
S.D.
0.006
0.007
TABLE 3: RESULT OF RECOVERY STUDY
Method
Level of recovery
80 %
I
100 %
120 %
80 %
II
100 %
120 %
80 %
III
100 %
120 %
*Average of five determinations, CIP-Ciprofloxacin, ORD-Ornidazole
RESULTS AND DISCUSSION: The proposed methods
were found to be simple, accurate and reproducible
for routine simultaneous estimation of ciprofloxacin
and ornidazole in combination. In 0.1 N sodium
hydroxide solution, ciprofloxacin and ornidazole
sho ed max at 272 nm and 319 nm respectively.
% Recovery* ± S.D.
CIP
100.02±1.14
100.60±0.96
100.79±1.29
100.44±0.88
100.75±0.72
100.23±1.73
99.44±0.89
100.25±0.62
100.73±1.63
ORD
99.83±1.06
100.75±1.01
100.45±0.88
100.18±0.97
100.25±1.42
99.34±1.29
100.18±1.55
100.25±1.42
101.34±1.29
As thei max differ more than 20 nm; simultaneous
equation method tried for their simultaneous
estimation in formulation. Also iso-absorptive point
was observed at 292 nm so the absorbance ratio
method was tried for simultaneous estimation of drugs
i fo ulatio at 7
max of ciprofloxacin) and
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Grewal et al., IJPSR, 2012; Vol. 3(8): 2716-2720
292 nm (iso-absorptive wavelength). The calibration
curve was linear over the concentration range 2.5 to
22.5µg/ml of Ciprofloxacin & Ornidazole. By observing
the validation parameters, all the methods were found
to be simple, specific, accurate and precise. Recovery
experiments indicated the absence of interferences
from the commonly encountered pharmaceutical
additives and excipients.
All the methods were successfully used to determine
the amounts of ciprofloxacin and ornidazole present in
the tablets. The results obtained were in good
agreement with the corresponding labelled amount.
Hence, all the three methods can be employed for the
routine analysis of these two drugs in combinations.
13.
14.
15.
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18.
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How to cite this article:
Grewal AS, Patro SK, Kanungo SK and Bhardwaj SK: Simultaneous
Spectrophotometric Estimation of Ciprofloxacin and Ornidazole in
Tablet Dosage Form. Int J Pharm Sci Res 2012; Vol. 3(8): 2716-2720.
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