Talanta, Vol. 42, No. I, pp.

105-108, 1995
Copyright ~3 1994 Elsevier Science Ltd
Pergamon 0039-9140(94)00224-X Printed in Great Britain. All rights reserved
0039-9140/95 $9.50 + 0.00

SPECTROPHOTOMETRIC DETERMINATION OF
DICLOFENAC SODIUM WITH METHYLENE BLUE

JULIO C. BOTELLO a n d GUADALUPE PI~REZ-CABALLERO*

Facultad de Estudios Superiores Cuautitl/m U.N.A.M., Secci6n de Quimica Analitica, Campo Uno. Av.
Primero de Mayo, s/N Cuautitlfin Izcalli, C.P. 54740, Estado de M6xico, M6xico

(Received 4 May 1994. Revised2 August 1994. Accepted4 August 1994)

Summary--A sensitive spectrophotometric method was established for the determination of Diclofenac
sodium (DS) with Methylene Blue (MB) as analytical reagent. It was found that DS reacts with an excess
of MB in the pH range 9.2-9.4, to form a chloroform-extractable blue ion-association complex. Good
agreement with Beer's law was found in the range of DS concentrations of 0.8~.4/ag/ml with a detection
limit of 0.37 pg/ml. The method was applied for the determination of DS in various tableted forms with
a good precision.

2-(2,6-dichloroanilino)Phenyl acetate (Diclofe- Research pH-meter, calibrated with buffer

nac sodium) (Fig. la) is a nonsteroidal anti-
inflammatory agent with potent activity and
outstanding tolerability in the treatment
of rheumatic diseases.L2 It has been measured
by a variety of analytical techniques such
as ultraviolet 3 and visible spectrophoto-
metry, 4"5 gas 6-8 and liquid chromatography 9-12
and nuclear magnetic resonance spectros-
copy. 13
Spectrophotometric methods provide sensi-
tive, precise and accurate measurements of suit-
able analytes and they offer practical and
economical advantages over other methods. Di-
clofenac sodium (DS) has been analyzed by
spectrophotometric methods using complexa-
tion with copper acetate 4 or Methylene Violet 5
followed by extraction, the latter being a more
sensitive method than the former. The aim of
the present work was to establish an improved
extraction and spectrophotometric method with
greater sensitivity for the determination of DS
using Methylene Blue (MB) (Fig. lb) in different
tableted dosage.
solution (pH 10 + 0.05).
Reagents
DS (standard drug), 99.46% purity, Voltaren
Retard and Flenaken tablets containing 100 mg
DS, and Voltaren tablets containing 50 mg DS
were analyzed. All other chemicals used were
analytical-reagent grade. Deionized water was
used to prepare all solutions and in all
experiments.
Solutions
A freshly prepared aqueous solution of the
pure drug (40 mcg/ml; 1.2573 x 10-4M) was
used as the standard solution for analytical
purposes. A 0.032% w/v (!.04 x 10-3M)
aqueous Methylene Blue solution was prepared
for these experiments. A m m o n i u m / a m m o n i a
buffer solutions covering the pH range from 8 to
11 were made by mixing equal volumes of 1.0M
ammonia and 1.0M a m m o n i u m chloride sol-
utions and adjusting the pH with sodium
hydroxide or hydrochloric acid.

EXPERIMENTAL
Apparatus
A Beckman DU-65 single beam spectropho-
tometer with 10 m m glass cells was used for the
absorbance measurements. The p H measure-
ments were made with a Corning Model 12
*Author to whom the correspondence should be addressed.
Procedure
Aliquots of DS solution were transferred into
a series of 125 ml separating funnels; then 5 ml
of MB solution and 1 ml of buffer (pH 9.4) was
added. The total volume of the aqueous phase
was adjusted to 10 ml with water. Chloroform
(10 ml) was added and the contents shook for
exactly 5 min, the separated chloroform layer
was collected in a 25 ml volumetric flask. A new

105
106 J. C. BOTELLO and G. Pi~REZ-CABALLERO

<•CHzCOONa
to 50 ml with deionized water and treated as
described above for the standard drug solution.
NH C l ~

C RESULTS AND DISCUSSION

(a)
N<
(CH3)aN
+ maximum at 653 nm against reagent blank,
(b)
Fig. 1. Structure of (a) Diclofenac sodium and (b) Methyl-
ene Blue.
10 ml volume o f chloroform volume was added
and shook for 5 min, this second extract was
combined with the first and this solution diluted
to the m a r k with chloroform. The absorbance of
the organic phase was measured against a re-
agent blank. The reagent blank was prepared
exactly like the procedure described above, but
in absence of DS.
Procedure of the assay of tab&ted dosage
An amount of tablet powder equivalent to 80
mg of anti-inflammatory agent was weighed
accurately and dissolved in water by magnetic
stirring. Filtration through a filter paper
(Whatman 40) was performed to remove insolu-
ble matter remaining. The solution was diluted
Absorption spectra
Diclofenac sodium reacted with MB in
aqueous solution in alkaline medium to form a
blue ion-association complex which was ex-
tracted into chloroform. Figure 2 shows the
absorption spectra of this complex and the
Cl- reagent blank. This complex has an absorption
N(CH3)=
hence this wavelength was used for all sub-
sequent measurements. Under the same exper-
imental conditions the reagent blank gave a
maximum absorption at 642 nm.
Optimum conditions for complex formation
In order to establish the optimum p H range,
DS was mixed with M B in aqueous solution
from p H 8 to 11, and the ion-associated com-
plex absorbance measured. Figure 3 shows that
the absorbance increases and reaches a maxi-
m u m and constant value at 9.2-9.5 p H range.
At p H values above 9.5 the absorbance decrease
was probably due to the formation of a new
ion-associated complex between MB and hy-
droxyl anion in alkaline medium since exper-
imentally, the reagent blank absorbance
increases. Hence, a pH of 9.4 was used in all the
subsequent experimental work. The shape of the
spectra and m a x i m u m position did not vary
with pH, so it was concluded that only one
complex was formed at this p H range.

0.9

A
B zq

S 0.6
O
R e i

B i i

A
N
C e" I

E 0.3 II

..... o°,"
° ° . . ° - °
0.0 I I
400 500 600 700
WAVELENGTH(nm)
Fig. 2. The absorption spectra of the blank reagent (---), and DS-MB ( ) complex formed in the
aqueous 0.05M NH+/NH3 buffer containing 5 x 10-4M MB and extracted with chloroform.
 Determination of Diclofenac sodium with Methylene Blue 107
0.7

A
B
$ 0.6
O
R
B
A
N
C 0.5
E

I I I I I
8.0 9.0 10.0 11.0 12.0
pH
Fig. 3. Variation in absorbance of the DS-MB complex, fonned in aqueous 0.05M NH+/NH3 buffer
containing 5 x 10-4M MB at different pH values (aborbances were obtained at 2653nm)"

The extent of the extraction of the ion-associ-
ated complex was found to be affected by the
MB concentration. To establish the optimum
MB concentration the solution absorbances
were plotted as a function of MB concentration
(Fig. 4). The absorbance of the system increased
in the range of 1-3.5 x 10-4M MB and the
absorbance was practically constant in a range
of 4-6 x 10-4M MB, hence a concentration of
5.0 x 10-4M MB was used as the optimum
concentration. The excess of MB in this system
was 25-200 times the concentration of DS.
Two extractions were necessary to achieve a
quantitative recovery of the complex. Ab-
sorbances of the separated extracts were stable
for at least 1 hr. A shaking time of 5 min
produced a constant absorbance. An ionic
strength of 0.025M was produced by the buffer.
Linearity
A calibration curve was obtained under opti-
m u m conditions (pH 9.4; 5 x 10-4M MB; ionic
strength 0.025M), absorbance responses were
linear in relation to the calculated concentration
of DS over the range 0.8-6.4 mcg/mi
(2.5 x 10-6-2.5 x 10-SM). The equation for
one representative calibration curve is:
A = ( - 0 . 0 1 9 1 + 0.0423) + (0.1789 + 0.0105)C,
where A and C correspond to absorbance
and DS calculated concentration (pg/mi),

1.3.

X I .-.'4
1.2'
A
B
S
1.1'
o
R
B
1.0,
A
N
C
E 0.9

0.8 ¸

0.7 I ! I I I i
0 1.0 2.0 3.0 4.0 S.0 6.0
[MB| (M X 10 -4)
Fig. 4. Variation in absorbance of the DS-MB complex, formed in aqueous 0.05M NH~/NH~ buffer
containing different concentrations of MB (absorbances were obtained at 2653,m).
108 J.C. BOTELLOand G. I~REZ-CABALLERO

Table 1. Comparison of sensitivity parameters of the differ- o f the m a t r i x effect over the absorbances
ent spectrophotometric methods for the determination of measurements.
Diclofenac sodium
Sandell Detection CONCLUSIONS
Analytical sensitivity limit
reagent (rag/cm 2) (mcg /ml) T h e p r o p o s e d m e t h o d is economical, simple,
Copper acetate 0.01026 1.00 rapid, precise a n d sensitive. The use o f M B as
Methylene Violet -- 72.00 a n a l y t i c a l reagent p r o v i d e s a fairly higher sensi-
Methylene Blue 0.006 0.37
tivity c o m p a r e d with o t h e r similar reagents such
as M e t h y l e n e Violet.
Table 2. Determination of Diclofenac sodium in different
tableted dosage Acknowledgements--We thank Dr Ignacio Gonzfilez for his
collaboration in this project. J. C. Botello acknowledges the
Labeled amount Amount found scholarship from CONACYT.
Drug (mg ) (rag)
Flenaken 100.00 101.45 + 0.42
Voltaren Retard 100.00 101.35 _+0.13 REFERENCES
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