Mikrochim.

Acta 137, 185±189 (2001)

New Colorimetric Methods for the Determination of Indapamide

and its Formulations

Hanna M. Saleh1; , Alaa S. Amin2 , and Magda El-Mammli1

1
Analytical Chemistry Department, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt
2
Chemistry Department, Faculty of Science, Benha University, Benha, Egypt

Abstract. Two simple and sensitive methods for the selectivity. Most analytical techniques for its determi-
determination of indapamide in pure and in dosage nation were concerning on GLC and HPLC methods
forms are developed. These methods are based on [5±13]. However, all these methods are costly, tedious,
the oxidation of indapamide with iron(III) in acidic time consuming and prior separation of the drug is
medium. The liberated iron(II) reacts with 1,10-phenan- required.
throline (Method A) and the ferroin complex is color- This paper describes colorimetric methods for the
imetrically measured at max 509 nm against reagent determination of indapamide which can be used in la-
blank. Method B is based on the reduction of Fe(III) by boratories where modern and expensive apparatus such
the drug. Iron(II) forms a colored complex (max as that required for GLC or HPLC are not available.
522 nm) with 2,20 -bipyridyl. Optimization of the experi- However, colorimetric methods are versatile and eco-
mental conditions is described. Beer's law is obeyed nomical particularly for developing countries. The pro-
in the concentration range 1:0±12 mg mlÿ1 and 4:0± posed procedures are also applicable for estimation
18 mg mlÿ1 for A and B, respectively. The apparent of indapamide in some pharmaceutical dosage forms
molar absorptivity and Sandell sensitivity for method without previous separation.
A is 3  104 L molÿ1 cmÿ1 and 0:0188 mg cmÿ2 , while
for method B is 2:3  104 L molÿ1 cmÿ1 and 0:0159
mg cmÿ2 . The detection and quanti®cation limits are Experimental
calculated. The developed methods are applied success- Apparatus
fully for the determination of indapamide in pure and
UV-Visible recording spectrophotometer Shimadzu-260 with 1.0 cm
in tablet form without interference from common ex- quartz cuvettes was used for all spectral measurements.
cepients.
Key words: Colorimetry; indapamide; pharmaceutical formula- Material and Reagents
tions; iron-phenanthroline; iron-bipyridyl complexes.
All chemicals and materials were of analytical grade and all
solutions were freshly prepared in double distilled water.
Indapamide [4-chloro-N(2-methyl-1-indolinyl)-3-sul-
phamoyl benzamide] is a diuretic agent having a potent 1. Pure indapamide and indapamide tablets (NatrilixR 2.5 mg) were
supplied by Les Laboratories Servier, France.
anti-hypertensive activity [1, 2]. In the UPS XXIV, 2. Preparation of iron(III)-o-phenanthroline reagent [14]. Mix
supplement, a liquid chromatographic assay for in- 0.198 g of 1,10-phenanthroline monohydrate (Sigma Chemical
dapamide is reported [1]. A review of the literature of Company, St. Louis, U.S.A.), 2.0 ml of 1.0 M HCl and 0.16 g of
the past two decades reveals that there are a few ferric ammonium sulphate dodecahydrate (Aldrich, Germany)
and diluted with water to the mark in 100 ml calibrated ¯ask.
spectrophotometric methods [3, 4] reported for the 3. Preparation of iron(III)-bipyridyl reagent (Aldrich, Germany):
assay of indapamide but lack both sensitivity and dissolve 0.16 g of 2,20 -bipyridyl in 2.0 ml of 1.0 M HCl and
0.16 g of ferric ammonium sulphate dodecahydrate and diluted to
 To whom correspondence should be addressed 100 ml with water in a 100 ml calibrated ¯ask.
186 H. M. Saleh et al.

4. Indapamide solutions: prepared by dissolving 100 mg of pure the reaction of indapamide with an iron(III)-o-phenan-
indapamide in 30 ml ethanol and completed to 100 ml with the
same solvent to obtain the standard solution of 1:0 mg mlÿ1 . throline or iron(III)-2,20 -bipyridyl reagent. The reaction
Working solutions were prepared by an appropriate dilution of proceeds through reduction of iron(III) ion to iron(II)
the stock standard solution. and subsequent formation of an intensive orange-red
coloration of the complex.
General Procedure The absorption spectra of the colored complex species
Aliquots of standard drug solution of indapamide ranging from 10± in the proposed methods show characteristic max at
120 mg for method A and from 40±180 mg for method B, were 509 and 522 nm using method A and B, respectively
transferred into a series of 10 ml calibrated ¯asks. 2.5 ml Fe(III)-o- (Fig. 1). The experimental conditions were established
phenanthroline or 3.0 ml Fe(III)-2,20 -bipyridyl reagent were added.
The ¯asks were then heated on a water bath at 70  C for 10 min by varying each parameter individually and observing
using method A and 20 min using method B. The contents of the the effect on the absorbance of the colored species. The
¯asks were cooled to room temperature (25  1  C) and the volume obtained complex is stable for at least 24 h.
is completed to the mark with distilled water. The absorbance was
measured at 509 nm for A and 521 nm for B against blank treated
similarly.
Effect of pH

Procedure for Tablets The pH adjustment is necessary especially in acidic

medium because the reaction is affected by the change
Twenty tablets were accurately weighed and powdered. An accu-
rately weighed quantity equivalent to 50 mg indapamide was of the pH in the range of 2.5±6.0. pH 3.5 is the
dissolved in 10 ml ethanol and transferred to a 100 ml calibrated optimum pH value of the ®nal assay solution for both
¯ask. The contents of the ¯ask was shaken for 10 min, and then methods A and B for complete oxidation of the drug
completed to the mark with the same solvent. The sample was
®ltered and the procedure was then continued as described above.
and obtaining a high color intensity and stability.

Results and Discussion Effect of Reagent Concentration

The methods A and B are based on the tris(o-phen- The addition of 2.5 ml of iron(II)-o-phenanthroline or
anthroline) or tris(2,20 -bipyridyl)-iron(II) chelate upon 3.0 ml of iron(III)-2,20 -bipyridyl reagent was required
to obtain a maximum and reproducible absorbance.
Smaller amounts give incomplete complex formation.
Therefore 2.5 ml of iron(III)-o-phenanthroline or 3.0 ml
of iron(III)-2,20 -bipyridyl reagent were used throughout
the experimental studies.

Effect of Temperature and Heating Time

Figures 2 and 3 show the effect of temperature and
heating time on the formation of the colored complex.
The reaction of indapamide with both reagents pro-
ceeds slowly at room temperature. Higher temperature
was used to accelerate the reaction. Maximum absor-
bance was obtained after heating (at 70  C) for about
10 min and 20 min for Fe2‡ -phen and Fe2‡ -bipyridyl
colored complexes, respectively. Therefore, develop-
ment times of 10 min at 70  C (method A) and 20 min
at 70  C (method B) were adopted in subsequent
investigations.

Sensitivity, Accuracy and Precision

Fig. 1. Absorption spectra of 1-Fe(III)-1,10-phenanthroline, 2-
Fe(III)-2,20 -bipyridyl before addition of 12 mg mlÿ1 and 3, 4 after Under the optimum conditions described above, Beer's
reduction with indapamide, respectively law holds well over the concentration range 1:0±12 mg
New Colorimetric Methods for the Determination of Indapamide and its Formulations 187

Fig. 2. Effect of temperature and heating time on

absorbance of the colored product from method
A. at 25  C; at 40  C; at 50  C; at
60  C; at 70  C

Fig. 3. Effect of temperature and heating time on

absorbance of the colored product from method
B. at 25  C; at 40  C; at 50  C; at
60  C; at 70  C

mlÿ1 using method A and 4:0±18 mg mlÿ1 of indapa- Table 1. Quantitative parameters for the proposed methods A and B
mide using method B. The apparent molar absorptivity Parameter Method A Method B
and Sandell sensitivity were found to be 3  104 max /nm 509 522
L molÿ1 cmÿ1 and 0:0188 mg cmÿ2 using method A, Beer's law/mg mlÿ1 1.0±12 4.0±18
whereas for method B were 2:3  104 L molÿ1 cmÿ1 Ringbom concentration range/mg mlÿ1 2.0±11 4.5±16.6
Molar absorptivity …"†=L molÿ1 cmÿ1 3  104 2:3  104
and 0:0159 mg cmÿ2 . The optimum concentration Sandell sensitivity/mg cmÿ2 0.0188 0.0159
ranges of indapamide that can be measured accurately, Detection limit/mg mlÿ1 0.25 0.90
as evaluated from Ringbom plot are 2.0±11 and 4:5± Quanti®cation limit/mg mlÿ1 0.83 3.1
Regression equation
16:6 mg mlÿ1 using methods A and B, respectively. The Intercept/a 0.003 0.007
relative standard deviation for nine replicate determi- Slope/b 0.078 0.055
nations of 7:0 mg mlÿ1 is 0.76 and 0.80% using method Correlation coef®cient/r 0.9997 0.9994
A and B, respectively. The regression equation using Relative standard deviation (Sr )/% 0.764 0.804

Fe2‡ -phen is A ˆ 0.003‡0.078 C, whereas using Fe2‡ -  A ˆ a‡bC where (C) is the concentration in mg mlÿ1 and (A) is
bipyridyl is A ˆ 0.007‡0.055 C (where C is the the absorbance unit.
concentration in mg mlÿ1 ). The standard deviations of
the absorbance measurements obtained from a series of The proposed methods are more sensitive compared
13 blank solutions for each reagent were calculated with Agrawal et al. [3] method using ammonium
(Table 1). The limits of detection (K ˆ 3) and of molybdate as reagent (molar absorptivity is 6:1  103
quanti®cation (K ˆ 10) of the methods were estab- L molÿ1 cmÿ1 with Sandell sensitivity of 0:059 mg
lished according to IUPAC de®nition [15]. cmÿ2 . Comparison of the recovery obtained with the
188 H. M. Saleh et al.

proposed methods with that of Agrawal et al. [3] formulated with indapamide. Therefore they could be
showed a high accuracy of the present methods. used easily for the routine analysis of pure indapamide
Moreover, the proposed methods could be used for and its dosage forms.
the routine determination of indapamide in pure or in The performance of the proposed methods was
dosage forms. assessed by calculation of the t-test (for accuracy) and
F-value (for precision) compared with the of®cial
method for 95% con®dence level with ®ve degrees of
Interferences
freedom [16]. The results showed that the t-values were
No interferences were observed in the determination 1.37 and 1.78 using method A and B, respectively,
of indapamide in presence of gum acacia, propylene whereas the critical value is 2.57. The F-values were
glycol, calcium lactate, calcium hydrogen phosphate, 2.85, and 3.41, respectively, while the critical value is
reserpin, zinc stearate, carboxymethyl cellulose, frac- 5.05. These results indicated that there was no signif-
tose, glucose, lactose, magnesium stearate and starch icant difference between the proposed and of®cial
when present in 120 fold molar excess. Also, there was methods.
no interference from the thermal and hydrolytic deg-
radation products of indapamide. In preliminary experi-
Conclusion
ments, these compounds were found not to reduce
iron(III) to iron(II) and therefore do not interfere in the The proposed method is simpler, less time consuming
determination. The results suggest that the methods and more sensitive than the of®cial method (based
would be useful for quality control of indapamide in on liquid chromatographic assay). Whereas the color
their pharmaceutical preparations. developments of Fe2‡ -phen and Fe2‡ -bipyridyl com-
plexes at room temperature required 60 and 90 min
using method A and B2 by raising the temperature to
Analytical Applications
70  C. The proposed methods are suitable for the
The proposed methods were successfully applied to determination of indapamide in pure and in dosage
determine indapamide in its dosage forms using the forms without interference from excipients such as
standard addition method in which variable amounts of starch and glucose or from common degradation
the pure drug were added to the previously analysed products, suggesting applications in bulk drug analysis.
portion of pharmaceutical dosage forms. Results are
shown in Table 2 and con®rm that the proposed meth-
References
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New Colorimetric Methods for the Determination of Indapamide and its Formulations 189

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