ISSN No:2321 8630, V 1, I 1, 2014

Journal Club for Pharmaceutical Sciences (JCPS)

Manuscript No: JCPS/RES/2014/22, Received On: 04/09/2014 , Accepted On : 22/09/2014, Published On: 21/01/2015

RESEARCH ARTICLE
Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric
Method Using Bratton Marshall Reagent
Rajan J Patel1, Shruti S Shah1, Chandani D Patel1, Rajnikant B Mardia1, B N Suhagia1
1

Department of Quality Assurance, Faculty of Pharmacy, Dharmsinh Desai University,

Nadiad, Gujarat, India.

ABSTRACT
A simple, rapid, sensitive and cost effective Colorimetric method has been developed for the
analysis of Nimesulide in bulk and its tablet formulation. In which Nimesulide was reduced,
diazotized and coupled with Bratton Marshall Reagent. The proposed method was validated
according to ICH guideline. Beers law was obeyed in the concentration range of 2-10 g/ml.
The correlation co-efficient was found to be 0.9936. The % recovery of Nimesulide was in
the range of 99.89-101.85 %. The coefficient of variance for Intraday and Interday precision
was found to be less than 2. Limit of detection and limit of quantification were 0.482g/ml
and 1.462g/ml respectively. The proposed method was applied for determination of
Nimesulide in the marketed formulation in which % assay was found to be 99.58 %. The
method was found to be, accurate, precise, repeatable and specific.
KEYWORDS
Nimesulide, Bratton Marshall Reagent, Colorimetric method.
INTRODUCTION

used

Nimesulide is chemically (4 - nitro - 2 -

rheumatoid arthritis and osteoarthritis);

phenoxy methanesulfonanilide) a non-

surgery and post traumatic acute pain and

steroidal anti-inflammatory drug

inflammation;

with

antipyretic and analgesic properties.It is

in

chronic

respiratory

arthritis

(such

dysmenorrhea;
tract

infection.[1-2]

as

upper
The

extensive review of literature revealed that

*Address for Correspondence:
Patel Rajan J,
Faculty of Pharmacy, Department of
Pharmaceutical Analysis,
Dharmsinh Desai University, Gujarat, India.
Email: [email protected]

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many analytical methods like fluorimetry

[3]

, spectrophotometry

thin

layer

[4-10]

, HPLC

chromatography

[11-14]
[15]

voltametry[16]

152

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
NHSO2CH3
Reagents and Solvents:
O

Methanol (LR Grade,Sulab Reagents,

Vadodara)
Zinc Dust (LR Grade, Sulab Reagents,
Vadodara)
Sodium

NO2

Figure

1:

Chemical

structure

of

Nimesulide
and

nitrite

(LR

Grade,

Sulab

Reagents, Vadodara)
Ammonium Sulphamate (LR Grade,

electrophoresis[17-18]have

capillary

been reported for estimation of Nimesulide

but not any colorimetric method reported
using Bratton Marshall Regent (N-1naphthyl

Sulab Reagents, Vadodara)

BMR reagent (LR Grade, SDFCL Ltd,
Mumbai)
Whatman filter paper no. 41 (Whatman
International Ltd., England)

ethylene

diaminedihydrochloride).

Instrument

A Shimadzu model 1800 double beam

So, it was thought of interest to develop

UV/Vis spectrophotometer with spectral

simple,

width of 1

precise

and

economical

0.2nm,

wavelength

spectrophotometric method with Bratton

accuracy of 0.1 nm and a pair of 10

Marshall Reagent (BMR) for estimation of

mm matched quartz cells

Nimesulide in bulk and its tablet dosage

form. The developed method was validated
for its linearity, accuracy, precision, limit
of

detection

(LOD)

and

limit

Analytical balance SHIMADZU AUX

220
Preparation of Standard Stock Solution

of

quantification (LOQ) according to the ICH

Nimesulide

(25mg)

was

weighed

guideline (Q2R1).

accurately and transferred into 25 ml

volumetric flask. Methanol (15ml) was

EXPERIMENTAL

added, shaken for 15 minutes and dilute up

Materials and Methods

to the mark with Methanol (1000 g/ml).

Nimesulide was procured as a gift sample

Working Standard Solution

from Dwarkesh health care, Vatva GIDC,

Standard stock solution (5 ml) was taken

Ahmedabad.

in 50 ml volumetric flask and dilute up to

Nimesulide

Tablet

(Nimeson 100mg)

50 ml with Methanol (100 g/ml).

procured from local market.

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153

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
Preparation of Sample Solution
dissolve and diluted up to the mark with
Twenty

tablets

(NIMESON)

were

distilled water.

weighed accurately and powdered. Powder

Preparation of BMR Solution (0.1

equivalent to 25 mg of Nimesulide was

%W/V)

transferred to 25 ml volumetric flask.

Methanol (15 ml) was added and sonicated
for 15 minutes and diluted up to 25 ml

BMR (0.1 g) was transferred into 100 ml

volumetric flask, dissolve and diluted up to
the mark with distilled water.

with Methanol. The solution was filtered

through Whatman filter paper No. 41.
Aliquot (5 ml) was diluted to 50 ml with
Methanol.
Preparation

of

Hydrochloric

Acid

volumetric flask.Zinc dust (0.5 gm) was

added and heated at 80C for 15 minutes.

acid

(0.85

ml)

was

transferred into 100 ml volumetric flask

and diluted up to the mark with distilled
water.
Preparation

Drug solution (100g/ml, 50ml) and 0.1N

HCl (50ml) were taken in the 100 ml

(0.1N)
Hydrochloric

Procedure

This solution was allowed to cool and

filtered through whatman filter paper and
diluted up to the mark with 0.1 N HCl. The
prepared drug stock solution (50g/ml).An
aliquot 2 ml was transferred in to 25 ml
of

Hydrochloric

Acid

(5%V/V)
Hydrochloric acid (5 ml) was transferred
into 100 ml volumetric flask and diluted
up to the mark with distilled water.

volumetric flask (4g/ml). HCl (5% V/V,

1ml) was added and keep aside for 5
minutes. Sodium nitrite (0.2 % W/V, 1ml)
was added and allowed to stand for 5
minutes. Ammonium sulphamate (0.5 %

Preparation of Sodium Nitrite (0.2

W/V, 1ml) was added and kept aside for 5

%W/V)

minutes. Then BMR (0.1 %W/V, 1ml) was

Sodium nitrites (0.2 g) was transferred into

100 ml volumetric flask, dissolve and
diluted up to the mark with distilled water.

added and kept aside for 5 minutes and

make up to mark with water. The
absorbance was measured at 559 nm.
Optimal Conditions

Preparation of Ammonium Sulphamate

In this method for the reduction of nitro

(0.5 %W/V)

group 80 C temperature required for 15

Ammonium sulphamate (0.5 g)

was

transferred into 100 ml volumetric flask,

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minutes than all the steps are performed at

laboratory environment.

154

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
Colorimetric Reaction:
Heating time for reduction of nitro group
was optimized at different time (0, 5, 10,
NHSO2CH3

0.5gm Zn dust +
0.1N HCl

NO2

80C for 15 min 15, 20, 25 and 30 minutes)

5% v/v HCl
0.2% w/v of NaN02

Nimesulide

0.5% w/v of Ammonium sulphamate

0.1% BMR

Optimization of Parameters for Method

Development
Optimization of Quantity of Zinc Dust
The Quantity of zinc dust was optimized
by taking different quantity of zinc dust
(0.3, 0.5, 0.7, 0.9 and 1.1gms). Other
procedures are same as mention above.
The solution was analyzed at 559 nm. The

Pink coloured complex at 559nm

Quantity of zinc dust related to highest

absorbance was selected as an optimized

interval. Other procedures are same as

condition.

mention above. The solution was analyzed

0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0

absorbance

absorbance

at 559 nm.

0.5

1.5

gm of zinc dust

0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
0

10

20

30

40

Heating time (minutes)

Fig. 1: Optimization of Heating Time

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Fig. 2: Optimization of Zinc dust

155

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
interval. Other procedures are same as
mention above. The solution was analyzed
mention above. The solution was analyzed

at 559 nm. The volume of0.2% NaNO2

at 559 nm.

related to highest absorbance was selected

as an optimized condition.

0.3

absorbance

absorbance

0.4
0.2
0.1
0
0

20

40

Heating time (minutes)

0.36
0.35
0.34
0.33
0.32
0.31
0.3
0

ml of NaNo2
Fig. 3: Optimization of Heating time

Fig. 5: Optimization of volume of 0.2%

NaNO2

Optimization of Volume of 5% HCl

The volume of 5% HCl was optimized by

Optimization

taking different volume of HCl (1, 2, 3, 4

Ammonium Sulphamate

and 5 ml). Other procedure is same as

mention above. The solution was analyzed
at 559 nm. The volume of 5% HCl related
to highest absorbance was selected as an
optimized condition.

The

volume

of

of

Volume

0.5%

0.5%

Ammonium

sulphamate was optimized by taking

different

volume

of

Ammonium

sulphamate (1, 2, 3, 4 and 5 ml). Other

procedure is same as mention above. The

volume of 0.5%Ammonium sulphamate

related to highest absorbance was selected
as an optimized condition.
0

ml of HCl
Fig. 4: Optimization of volume of 5%
HCl
Optimization of Volume of 0.2% NaNO2

0.36
0.35
0.34
0.33
0.32
0.31
0.3

Abs.

absorbance

solution was analyzed at 559 nm. The

0.35
0.34
0.33
0.32
0.31
0.3

2
4
6
ml of Ammonium sulphamate

The volume of 0.2% NaNO2was optimized

by taking different volume of NaNO2 (1, 2,
3, 4 and 5 ml). Other procedure is same as

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Figure 6: Optimization of volume of

0.5% Ammonium sulphamate

156

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent

Fig. 7: Overlain spectra for of Nimesulide 2-10 g/ml

Absorbance Vs Conc.
1.2

y = 0.1003x - 0.0109
R = 0.9936

Abs

0.8
0.6
0.4
0.2
0
0

10

12

Conc. (g/ml)

Fig. 8: calibration curve of the colorimetric method for Nimesulide

Validation of the proposed method
The proposed method was validated

solution was allowed to cool and filtered it

according to the International Conference

and dilute up to the mark with 0.1 N HCl.

on Harmonization (ICH) Q2R1 guideline.

The

RESULT AND DISCUSSION

Linearity
Drug solution (100g/ml, 50ml) and 0.1N
HCl (50ml) were taken in the 100 ml
volumetric flask. Zinc dust (0.5 gm) was
added and heated for 15 minutes. This

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prepared

drug

stock

solution

(50g/ml).An aliquot (1, 2, 3, 4 and 5ml)

were transferred in to series of 25ml
volumetric flasks. HCl (5% V/V, 2ml) was
added in each flask and keep aside for 5
minutes. Sodium nitrite (0.2 % W/V, 2ml)
was added in each flask and allowed to
stand for 5 minutes.

157

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
Ammonium sulphamate (0.5 % W/V, 2ml)
The intra-day and inter-day precision of
was added in each flask and kept aside for

the proposed method was done by

5 minutes. Then BMR (0.1 %W/V, 2ml)

analysing the corresponding responses

was added in each flask and kept aside for

three times on the same day and on three

5 minutes and mark up to with water. The

different days over a period of one week

absorbance was measured at 559 nm.

for three different concentrations over the

calibration range of Nimesulide (4, 6 and

Accuracy (% Recovery)
The

accuracy

of

8g/ml). The results were reported in

the

method

was

performed by calculating % recovery of

terms of relative standard deviation (%

RSD).

Nimesulide by the standard addition

method. Known amounts of standard

Table 2: Results of Intra-day precision

solutions of Nimesulide were added at

and Inter-day precision study

50%, 100% and 150% levels to prequantify.

N
(g/ml)

Intra-day
precision (n = 3)

Inter-day
precision (n = 3)

Mean
S.D.

%RSD

Mean
S.D.

%RSD

0.358
0.0015

0.42

0.362
0.0035

0.96

0.581
0.0057

0.99

0.586
0.0040

0.69

0.790
0.0015

0.19

0.800
0.0036

0.45

At each level of the amount, three

determinations

were

performed.

The

amount of Nimesulide was estimated by

using

regression

equation.

The

Recovery was found to be in the range of

99.63-101.95%.
Table 1: Results of drug recovery data

N=Nimuslide

of Nimesulide
Repeatability
Amount Amount Amount
taken
added recovered

Recovery
S.D %
(n = 3)

%
RSD

by repeated scanning and measurement of

100.0

100.72
1.49

1.48

100.75

100.01
0.77

0.78

99.63

101.95

99.89
0.86
101.95
0.90

The precision of the methods was assessed

the absorbance of solutions (n = 6) of

Nimesulide (6 g/ml) without changing
the parameters for the methods.

0.89
0.98

Amounts in g

The % RSD was found to be 0.15 that

confirmed the repeatability of the proposed
method.

Precision

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158

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
Robustness study was carried out by
Table 3: Result of Repeatability study of
Nimesulide

changing the wavelength by 2 nm. The

Drug

Absorbance

0.582

0.581

0.580

0.582

0.580

0.581

Mean

0.581

S.D.

0.00089

% RSD

0.15

Limit

of

Detection

and

% RSD was found to be 0.52.

Analysis of Tablet Dosage Form
The proposed UV spectrophotometric
method was successfully applied for
determination of Nimesulide in tablet
dosage

claim.
Limit

of

Limit of detection (LOD) and the limit of

were

percentage

of

calculated

using the standard deviation of response

() and slope (S) of the calibration curve
LOD = 3.3 x /S
LOQ =10 x /S
So, the limit of detection and limit of
quantification were found to be 0.482
g/ml and 1.462 g/ml respectively.

No

any

interference

due

to

excipients was found in the assay results.

Each

(LOQ)

The

Nimesulide was found corresponding label

Quantification

quantification

form.

result

was

average

of

determinations.
Table 4: Results of assay of marketed
formulation of Nimesulide
NIMESON
tablet
1
2
3

Label
claim (mg)
100
100
100

Amount
Recovered
(mg)
98.5
99.83
100.43

Mean
Standard deviation
Relative standard deviation

% Amount
Recovered
(n = 3)
98.5
99.83
100.43
99.58
0.98
0.99

Sandells Sensitivity[19]

CONCLUSION

It is the concentration of the analyte (in

The proposed colorimetric method for

g/ml) which gives absorbance of 0.001 in

Nimesulide was found to be specific as

a cell of path length 1 cm. and is expressed

there was no interference of any excipients

as g/cm2 here the value of sandells

and impurities. The proposed method was

sensitivity was calculated to be 0.1625

found to be simple, accurate, precise,

g/cm2/0.001.

robust, repeatable and cost effective.

Robustness

Hence, it can be used successfully for the

routine analysis of Nimesulide and in
pharmaceutical dosage forms.

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159

Analysis of Nimesulide in Bulk and Tablet Formulation by Colorimetric Method Using Bratton Marshall Reagent
ACKNOWLEDGEMENT
spectrophotometric method for the
The Authors are thankful to Mr. Arpit
Patwari

for

valuable

guidance

and

Dharmsinh Desai University and Faculty

of Pharmacy for providing the research
facilities and also thankful to Dwarkesh
Health carefor

providing

the

graties

sample of Nimesulide for research work.

estimation

of

nimesulide.

Drug

Development and Industrial Pharmacy,

26(2), 229-234.
7. Nguyen, T.M.T., Nguyen, D.T. (2003).
Determination of nimesulide in tablets
by

second-derivative

UV

spectrophotometric method. Tap Chi

Duoc Hoc, 4, 16-18.

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HOW TO CITE THIS ARTICLE

Patel, R. J., Shah, S. S., Patel, C. D., Mardia, R. B., Suhagia, B. N. (2014). Analysis of
nimesulide in bulk and tablet formulation by colorimetric method using bratton marshall
reagent. Journal Club for Pharmaceutical Sciences (JCPS), 1(I), 152-161.

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161