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Three Spectrophotometric Methods for

the Determination of Oxomemazine
Hydrochloride in Bulk and in Pharmaceutical
Formulations Using Bromocresol Green,
Congo Red, and Methyl Orange
a b b
Alaa S. Amin , Mohamed A. El‐Mossalamy , Hamada M. Killa & Amr L.
b
Saber
a
Faculty of Science, Chemistry Department , Benha University , Benha,
Egypt
b
Faculty of Science, Chemistry Department , Zagazig University , Zagazig,
Egypt
Published online: 29 Jan 2008.

To cite this article: Alaa S. Amin , Mohamed A. El‐Mossalamy , Hamada M. Killa & Amr L. Saber (2008)
Three Spectrophotometric Methods for the Determination of Oxomemazine Hydrochloride in Bulk and in
Pharmaceutical Formulations Using Bromocresol Green, Congo Red, and Methyl Orange, Analytical Letters,
41:1, 80-89, DOI: 10.1080/00032710701746808

To link to this article: http://dx.doi.org/10.1080/00032710701746808

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 Analytical Letters, 41: 80–89, 2008
Copyright # Taylor & Francis Group, LLC
ISSN 0003-2719 print/1532-236X online
DOI: 10.1080/00032710701746808

PHARMACEUTICAL ANALYSIS

Three Spectrophotometric Methods for the

Determination of Oxomemazine
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Hydrochloride in Bulk and in

Pharmaceutical Formulations Using
Bromocresol Green, Congo Red, and Methyl
Orange

Alaa S. Amin,1 Mohamed A. El-Mossalamy,2 Hamada M.

Killa,2 and Amr L. Saber2
1
Faculty of Science, Chemistry Department, Benha University, Benha,
Egypt
2
Faculty of Science, Chemistry Department, Zagazig University,
Zagazig, Egypt

Abstract: Three simple, sensitive, and highly accurate spectrophotometric methods

have been developed for the determination of oxomemazine hydrochloride (OXO-
HCl) in bulk and in pharmaceutical formulations. These methods are based on the
formation of yellow ion-pair complexes between the examined drug and bromocresol
green (BCG), congo red (CR), and methyl orange (MO) as reagents in universal buffer
solution of pH 3.0, 5.5, and 3.5, respectively. The formed complexes were extracted
with chloroform and measured at 413, 495, and 484 nm, respectively for the three
systems. The best conditions of the reaction were studied and optimized. Beer’s law
was obeyed in the concentration ranges 2.0 – 18.0, 2.0 – 14.0, and 2.0 – 16.0 mg ml21
with molar absorptivity of 4.1
104, 1.1
104, and 3.5
104 mol21cm21, for the
BCG, CR, and MO methods, respectively. Sandell’s sensitivity, correlation coefficient,
detection, and quantification limits are also calculated. The proposed methods have
been applied successfully for the analysis of the drug in pure and in its dosage
forms. No interference was observed from common pharmaceutical excipients and
additives. Statistical comparison of the results with those obtained by HPLC method

Received 4 October 2007; accepted 1 November 2007

Address correspondence to Amr L. Saber, Faculty of Science, Chemistry Depart-
ment, Zagazig University, Zagazig, Egypt. E-mail: amrlotfy2000@yahoo.com

80
 Determination of Oxomemazine Hydrochloride in Bulk 81

shows excellent agreement and indicates no significant difference in accuracy and

precision.

Keywords: Spectrophotometric, oxomemazine HCl determination, ion-pair

complexes, dosage forms

INTRODUCTION
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Oxomemazine, a phenothiazine derivative, is an antihistamine used for the

symptomatic relief of hypersensitivity reaction. It is also an ingredient of
compound preparations for the symptomatic treatment of coughs and the
common cold. It is given orally in doses equivalent to 10 to 40 mg of oxome-
mazine daily. Oxomemazine may also be administered rectally in form of sup-
positories. Oxomemazine hydrochloride (OXO-HCl) has been used similarly
by mouth. It is chemically known as N,N,b-trimethyl-3-(10H-phenothiazin-
10-yl)-propan-1-amine-5,5-dioxide (Jacob et al. 1961), (Fig. 1).
The assay of the drug in pure and dosage forms is, as far as we know, not
official in any pharmacopoeia, and therefore requires much more investi-
gation. The different analytical methods that have been reported for its deter-
mination include HPLC (Hoogewijs and Massart 1984; Hewala 1994), GC
(Hans and Karl 1988; Robert and Berges 1970), and TLC (Kelber 1970)
methods.
Although the visible spectrophotometric methods are the instrumental
methods of choice commonly used in industrial laboratories because of
their simplicity, selectivity, and sensitivity, in the literature, only few spectro-
photometric (El-Didamony 2005; Coclers and Bontemps 1985; Zivanov-
Stakic and Deric 1979; Jean et al. 1967), IR (Barbe 1972; Maria and Maria
1979), microcalorimetric titration (Gerhard et al. 2007), volumetric titration
(Zivanov-Stakic and Djceric 1977), LC-ES1-MS (Saber et al. 2007), and sep-
aration of drug enantiomers by capillary electrophoresis (Koppenhoefer et al.
1995; Lin et al. 1996) methods have been reported. The aim of the present
work is to develop three simple, rapid, accurate, and sensitive extractive spec-
trophotometric methods for the determination of OXO-HCl. The methods are

Figure 1. Chemical structure of oxomemazine hydrochloride.

 82 A. S. Amin et al.

based on ion-pair complexes of the OXOHþ cation with dye anions such as
bromocresol green (BCG), congo red (CR), and methyl orange (MO) and sub-
sequent extraction into chloroform under the reaction condition used.

EXPERIMENTAL

Material and Reagents

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All chemicals and reagents used were of analytical grade and all solutions were
prepared in doubly distilled water. Pure OXO-HCl was obtained from October
Pharma, Cairo, Egypt. A stock solution of OXO-HCl (100 mg ml21) was
prepared by dissolving 10 mg of pure drug in least amount of doubly
distilled water, transferred into a 100 ml measuring flask, and diluted with
water up to the marking. Working solutions of lower concentrations were
prepared by serial dilution. Solutions of 5.0
1024, 2.0
1023, and
1.0
1023 M of BCG, CR, and MO, respectively, were prepared by dissolving
accurately weighted acid dyes in a few drops of ethanol and then diluted them,
separately, to the marking with water in 100 ml measuring flasks. A series of
Britton-Robinson (B-R) universal buffer solutions were prepared by
following the standard method (Britton 1952).

Instrumentation

Spectral and absorbance measurements were made with a JASCO 530 UV-Vis. and
Perkin-Elmer Lambda 3B spectrophotometer with 10 mm quartz cells. The pH of
solutions was checked using an Orion Research Model 601 A/digital ionlyser.

Procedure of Calibration Curve

Aliquots of OXO-HCl containing up to 180 mg ml21 were transferred into a

series of 50 ml separating funnels. 2.5 ml (5.0
1024 M), 3.6 ml
(2.0
1023 M), and 2.6 ml (1.0
1023 M) of BCG, CR, and MO, respect-
ively, were added and then the total volume of the aqueous phase was
adjusted to 10 ml by addition of the optimum pH of buffer solution
(Table 1). Finally, 10 ml of chloroform were added to each funnel and the
contents were shaken well for 2.0 min. The two phases were allowed to
separate and the chloroformic layer was dried by running through
anhydrous sodium sulfate. The absorbance of the yellow-colored species
was measured after standing for 5.0 min at lmax as recorded in (Table 1), in
each case against a reagent blank similarly prepared. A calibration graph of
the absorbance versus the concentration of the drug was plotted.
 Determination of Oxomemazine Hydrochloride in Bulk 83

Table 1. Quantitative parameters for determination of OXO-HCl using BCG, CR,

and MO

Parameters BCG CR MO

pH 3.0 5.5 3.5

Extracting solvent chloroform chloroform chloroform
lmax 413 495 484
Molar ratio (OXO-HCl:dye) 1:1 2:1 1:1
pK 4.24 5.25 4.50
Beer’s law limits (mg ml21) 2.0 – 18 2.0– 14 2.0– 16
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Ringbom range (mg ml21) 1.6 – 15.4 1.8– 13.2 1.6– 14.2
Molar absorptivity (L mol21 cm21) 4.1
104 1.1
104 3.5
104
Sandell’s sensitivity (ng cm22) 9.1 34 10.5
Range of error (%) 20.44:0.58 20.31:0.34 20.71:0.55
Regression equationa
Intercept 0.010 0.080 0.036
Slope 0.111 0.029 0.095
Correlation coefficient (r) 0.9992 0.9911 0.9997
t-value (2.56)b 1.19 1.56 1.33
F-value (5.05)b 2.48 2.93 2.67
pK 4.25 5.19 4.4
a
A ¼ a þ bC, where C is the concentration in mg ml21.
b
Values in parentheses are the theoretical values for t- and F-values at 95% confi-
dence limits and five degrees of freedom.

Procedure for Dosage Forms

Take a volume of syrup solution containing 25 mg of each of OXO-HCl

and make the volume 250 ml with distilled water, Filter if necessary
and further dilution was carried out with water. The general procedure
described above was used for the determination of OXO-HCl concen-
tration using blank prepared in the same manner except for the drug. A
standard additions technique was also used to confirm the accuracy and
precisions.

RESULTS AND DISCUSSION

OXO-HCl forms ion-pair complexes in acidic medium with acidic dyes

such as BCG, CR, and MO, and these complexes are quantitatively
extracted into chloroform. The absorption spectra of the ion-pair
complexes were measured in the range 300– 600 nm against the blank
solution similarly prepared (Fig. 2). The ion-pair complexes show
maximum absorbance at lmax for each three systems as recorded in
(Table 1).
 84 A. S. Amin et al.
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Figure 2. Absorption spectra of OXO . Hþ-acid dye complex extracted in

chloroform.

Optimization of the Reaction Conditions

The optimum conditions for quantitative estimation of the drug were estab-
lished by a number of preliminary experiments.

Effect of pH

The influence of pH on the ion-pair formations of OXO-HCl with various dyes

has been studied using different pH values of universal buffers. It is evident
that the maximum color intensity and maximum absorbance were found in
universal buffer of pH 3.0, 5.5, and 3.5 with BCG, CR, and MO, respectively
as shown in (Fig. 3). OXO-HCl contains a tertiary amino group, which is pro-
tonated in acid medium. The protonated OXO-HCl forms ion-pairs with
anionic dyes, which are quantitatively extracted into chloroform. The
possible reaction mechanisms are proposed as follows.

OXO - HCl þ BCG  Na ! ðOXO  HÞþ  ðBCGÞ þ NaCl

2OXO - HCl þ CR  2Na ! ðOXO  HÞþ
2  ðCRÞ

þ 2NaCl
þ 
OXO - HCl þ MO  Na ! ðOXO  HÞ  ðMOÞ þ NaCl

Effect of Dye Concentration

The effect of the dye concentration on the intensity of the color developed at
the selected wavelength and constant OXO-HCl concentration was examined
 Determination of Oxomemazine Hydrochloride in Bulk 85
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Figure 3. Effect of pH on the absorbance of OXO.Hþ-acid dye.

using different amounts of the reagents. The results indicated that the
maximum absorbance in each case was found with 2.5, 3.6, and 2.6 ml of
BCG, CR, and MO reagents, respectively, which are used for ion-pair-
formations throughout the experiment.

Choice of Organic Solvent

A number of organic solvents such as chloroform, dichloromethane, carbon

tetrachloride, benzene, and toluene were examined for extraction of the
formed ion-pair complexes in order to provide an applicable extraction
procedure. Chloroform was found to be the most suitable solvent for extrac-
tion of colored complexes yielding maximum absorbance intensity. It was
also observed that only one extraction was adequate to achieve a quantitative
recovery of the complex and the shortest time to reach the equilibrium
between both phases.

Phase Ratio

The ratio of aqueous to organic phase was ineffective and the ratio 1:1 was
chosen for extraction of the colored species. It was also noticed that the
order of addition of the reagents had neither an effect on the absorbance nor
the color of the complexes.

Effect of Shaking Time

Shaking for 1.0 –5.0 min, provided a constant absorbance and hence, 2.0 min
was used as an optimum shaking time throughout the experiment. The ion-pair
 86 A. S. Amin et al.

complexes were quantitatively recovered in one extraction step only and were
stable for at least 24 h without any change in color intensity. Although the ion-
pair complexes were formed instantaneously, constant absorbance readings
were obtained after not less than 5 min of standing at room temperature
(25 + 28C).

Composition of Ion-pair Complexes

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The composition of ion-pair complexes was established by Job’s method of

continuous variations using variable dye and OXO-HCl concentrations. The
results indicated that 1:1 (OXO-BCG & OXO-MO ion-pairs) and 2:1
(OXO-CR ion-pairs) were formed through the electrostatic attraction
between the positive protonated drug and the anion of the studied dyes. The
extraction equilibrium can be represented as follows:

OXOHþ 
ðaq:Þ þ Dðaq:Þ ! OXOHþ 
ðaq :ÞDðaq:Þ ! OXOHþ D
ðorg:Þ

where OXOHþ and D2 represent the protonated OXO-HCl and the anion of
the dye, respectively. The subscript (aq.) and (org.) refer to the aqueous and
organic phases. The shapes of the resulting curves indicate that the ion-pair
are labile. Consequently, a large excess of reagent must always be used to
enhance the formation of the complexes. The spectrophotometric methods
that are usually applied to determine the stoichiometric ratio of the
complexes can also be used for the determination of their stability constant
in solution. The values of the stability constant showed that the complex of
the studied drug with congo red is more stable than methyl orange and bromo-
crysol green reagents (Table 1).

Analytical Data

The Beer-Lambert law limits, molar absorptivity, Sandell’s sensitivity,

regression equations, and correlation coefficients obtained by linear square
treatment of the results are given in Table 1. In order to determine the
accuracy and precision of the three methods, three different concentrations
of OXO-HCl were prepared and analyzed in six replicates. Therefore, the
ion-pair formation was successfully used for the determination of oxomema-
zine. Percentage relative standard deviation (RSD %) as precision and percen-
tage relative error (RE %) as accuracy of the proposed methods were
calculated (Table 2). These results of precision and accuracy showed that
the proposed methods have good repeatability and reproducibility. The
percent relative standard deviation (RSD %) and recoveries were found to
vary over the range of 0.018 – 0.143% and 99.30 –100.58%, respectively.
 Determination of Oxomemazine Hydrochloride in Bulk 87

Table 2. Evaluation of accuracy and precision for the proposed methods

Taken
Procedure (mg ml21) Recoverya (%) RSD (%) RE (%)

MO 8.0 99.44 0.113 20.56

10.0 100.56 0.109 0.55
12.0 99.30 0.143 20.71
CR 8.0 99.68 0.063 20.31
10.0 99.91 0.018 20.09
12.0 100.34 0.068 0.34
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BCG 8.0 99.56 0.088 20.44

10.0 99.70 0.060 20.30
12.0 100.58 0.116 0.58
a
Average of six determinations.

Interference Studies

The effect of common excipients and other substances were tested for possible
interference in the assay. It was observed that talc, glucose, starch, sulfate,
dextrose, acetate, phosphate, and magnesium stearate did not interfere with
the determination at the levels found in dosage forms. However, the drug
content from syrup was extracted into chloroform, which eliminates the
common excipients found in the drug formulations.

Analytical Applications

The proposed methods have been successfully applied to the determination

of OXO-HCl in formulations. The results obtained by the proposed methods
were compared by official method (Table 3). For further confirmation, the
standard addition method was applied to test the reliability and recovery
of the proposed methods, since the ion-pair complexes are stable for at
least 24 h. The high percentage recoveries indicate that the excipients in
pharmaceutical dosage forms of OXO-HCl such as talc, glucose, starch,
lactose, sulfate, dextrose, and acetate, phosphate, and magnesium stearate
were not found any exhibit and any interference in the analysis of
OXO-HCl.
The results obtained for the proposed methods were compared with those
obtained using the HPLC method (HPLC Method of 6th October Pharm
2003). The calculated student’s t-values and F-values did not exceed the
theoretical ones at 95% confidence level (Miller and Miller 1993).
Therefore, there is no significant difference between the proposed methods
and HPLC methods.
 88
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Table 3. Determination of OXO-HCl in its formulations using the proposed and HPLC methods

Founda
Taken (mg Added (mg
Sample Method Manifested by ml21) ml21) Official Proposed Recovery (%) S.D. (%)

Toplexil BCG Octoberb 3.0 2.95 2.98 99.33 0.53

Pharmab 3.0 5.92 6.01 100.17 0.62
6.0 8.90 9.05 100.56 0.98
9.0 12.10 12.07 100.58 1.13
12.0 15.20 14.95 99.67 0.57
15.0 18.25 17.90 99.44 0.75
CR 2.0 1.96 2.01 100.50 0.83
2.0 3.95 4.02 100.50 0.65
4.0 5.95 6.00 100.00 0.36
6.0 8.05 7.96 99.50 0.49
8.0 10.10 9.95 99.50 0.78
10.0 11.85 12.05 100.42 0.97
12.0 14.15 13.92 99.43 0.82
MO 2.5 2.53 2.48 99.20 0.51
2.5 5.07 4.96 99.20 0.67
5.0 7.58 7.48 99.73 0.71

A. S. Amin et al.
7.5 10.15 10.05 100.50 0.43
10.0 12.40 12.55 100.40 0.82
12.5 15.20 14.90 99.33 0.99
a
Average of six determinations.
b
October Pharma, Cairo, Egypt.
 Determination of Oxomemazine Hydrochloride in Bulk 89

CONCLUSIONS

In the present study, OXO-HCl was determined successfully in bulk drug and its
pharmaceutical preparations. The maximum color development of ion-pair
complex formations, are completed immediately after all reagents were added
and thus, all the measurements were made in 5.0 min after standing the
extracted solutions in all the experiments. The commonly used additives and
excipients in pharmaceutical dosage forms of OXO-HCl did not interfere in
the analysis. The proposed methods are simple, sensitive, and reproducible and
can be used for routine analysis of OXO-HCl in pure form and in formulations.
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