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! ABSTRACT
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| The enhanced clinical use of sulfadoxine necessitated the study of new methods
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I for its determination in quality control laboratories. New sensitive visible
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| spectrophotometric methods are developed for the determination of sulfadoxine.
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| Proposed methods are based on the reaction of drug with aldehyde followed by cobalt(II)
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| chloride in acidic medium. The reaction of aldehyde with drug results in the formation of
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I Schiff base which act as a ligand for the formation of complex with Co(II). Developed |
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| methods are successfully applied to the pharmaceutical formulation. |
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Sulfadoxine
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6.1. DRUG PROFILE
Sulfadoxine is chemically 4-amino-iV-(5,6-dimethoxypyrimidin-4-yl)benzene-l-
sulfonamide belonging to the class of drug known as sulfanilamides. It is mainly used for
the treatment or prevention of malaria and also used as anti-infective agent [1].
Sulfadoxine has microbial activity similar to that of sulfadiazide. However, it is
principally used for the suppression of malaria caused by chloroquine-resistant
plasmodium falciparum [2]. Sulfadoxine helps to inhibit the enzyme dihydropteroate
synthetase which is an enzyme necessary in the conversion of /?-aminobenzoic acid to
folic acid [3], Folic acid is essential for synthesis and methylation of DNA which is vital
to cell growth in plasmodium falciparum [4]. In such a case because of the nutrient
lacking, parasite find difficulty in reproducing. Sulfadoxine is an ultra-long-lasting
sulfonamide often used in combination with other drugs, to treat or prevent various
infections in livestock, respiratory, urinary tract, and malarial infections [5].
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the maximum absorption at 505 nm for amodiaquine and at 470 nm for sulfadoxine. The
proposed method was applicable for the determination of these drugs in pharmaceutical
formulations.
Syed et al, [13] contributed new spectrophotometric method for the
determination of sulfa drugs namely, sulfanilamide, sulfadoxine and sulfamethoxazole.
The methods were based on the interaction of diazotized sulfa drugs with cardanol to
produce a yellow colored product with a maximum absorption at 415 nm. The color
developed was stable up to 24 hrs,
Balyejusa et al, [14] reported first derivative spectrophotometric method for the
simultaneous separation of sulfadoxine and pyrimethamine. Method was achieved by
applying zero-crossing. Linear calibration graphs of 1 st-derivative values at 273 and 240
nm for sulfadoxine and pyrimethamine respectively with negligible intercepts were
obtained. The authors recommend that it becomes a favorable alternative to the expensive
HPLC method.
Sastry et al., [15] proposed two new spectrophotometric methods for the
determination of sulfaethidole and sulfadoxine in pure samples and in dosage forms. The
1st method was based on the reaction of the primary amino group of the drugs with the
quinonimine produced in situ from metol-Cr(VI) to give a colored charge-transfer
complex with an absorption maximum at 520 nm; the 2 nd method involves the formation
of a colored azo dye (X,max 415 nm) through the coupling of phloroglucinol with each of
the diazotized drugs.
Raghuveer et al, [16] reported a sensitive spectrophotometric method for the
determination of sulfadoxine was based on the reaction with
4- dimethylaminocinnamaldehyde in orthophosphoric acid medium to form a stable and
sensitive chromogen.
Mohamed et al, [17] developed a simple spectrophotometric method for the
determination of 15 sulfonamides in bulk and in dosage forms. The method was based on
the interaction of p-benzoquinone with sulfonamides in 0.1 M HC1. The resulting
chromophore was measured at 500 nm. The effects of different variables on color
development were established.
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Rao et al, [18] described spectrophotometric method for the determination of
sulfadoxine and sulfalene in pharmaceutical dosage forms by using sodium
l,2-naphthoquinone-4-sulfonate. Developed colored chromogens were measured at
425-430 nm.
Rao et al, [19] reported spectrophotometric method for the determination of
pindolol and sulfadoxine in pharmaceuticals. Method was based on the treatment of the
drugs with Na2CC>3 soln. followed by reaction with Folin-Ciocalteu reagent and
measurement of absorbance at 765 nm.
Parimoo [20] reported differential spectrophotometric method for the
simultaneously analysis of pyrimethamine and sulfadoxine in syrups by a based on the
measurement of the absorbance in various media at 220-320 nm.
Rao et al, [21] proposed simple spectrophotometric method for the determination
of sulfadoxine and sulfalene based on reaction with metol at pH 2.5 using NaI04 as
oxidant. The chromogens formed had absorption maximum at 520 nm. It was found that
pyrimethamine and trimethoprim present in combination with these drugs did not
interfere.
Rao et al, [22] reported new spectrophotometric method for the determination of
sulfadoxine and sulfalene in combined dosage forms by reaction with o-chloranil at pH
9.0 and measurement of the absorbance of the chromogens at 525 nm.
Quite a few visible spectrophotometric methods have been developed for the
quantification of sulfadoxine in pharmaceuticals. Therefore, developing a selective and
sensitive methods using visible spectrophotometry is of paramount importance. Thus, aim
of the present work is to develop new and simple spectrophotometric methods for the
determination of sulfadoxine.
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6.4. EXPERIMENTAL
6.4.1. Apparatus
All solutions were prepared with double distilled water. Chemicals used were of
analytical reagent grade. Solutions of p-dimethylaminobenzaldehyde (PDAB) and
vanillin (VA) (5 * 10'4 M) were prepared in ethanol and cobalt(II) chloride
(1.5403 x 10"4 M) was prepared with water. About 0.1 g of pure sulfadoxine (SFD)
weighed accurately and dissolved in 5 mL of (2 M) hydrochloric acid and diluted to 100
mL with ethanol (1000 pg mL"1).
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6.4.33. Assay offormulation
The proposed method was applied for the determination of SFD in tablet
formulation. Tablet weight equivalent to 750 mg of SFD (Reziz forte) were crushed
thoroughly in a mortar and dissolved in 20 mL of 2 M hydrochloric acid and diluted to
100 mL by using ethanol. The solution was filtered through Whatmann No.l filter paper
and diluted quantitatively with ethanol to obtain a suitable concentration for the analysis.
Different aryl aldehydes are tested, out of which PDAB and VA gave stable green
colored complexes (Table 6.1). The color stability of these complexes may be due to the
presence of electron donating groups in PDAB (-NMe2) and VA (-OH and -OMe) which
helps in the formation of Co(II) complexes. Other aryl aldehydes do not form any colored
complexes.
The reaction is carried out at room temperature. It has taken around 5 min for the
complete color development. After the color development absorbance of the complex is
found to be constant up to 6 hrs. The series of solutions containing 60 pg mL'1 of drug
solution is taken and various amount of reagent ranging from 0.5-2.00 mL is added. It is
found that 1 mL of reagent is sufficient to form Schiff base ligand with drug and 0.5 mL
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of metal solution is found to be optimum to form stable complex with the ligand in both
the cases.
In order to study the accuracy and precision of the proposed methods, three
concentrations of pure SFD within the linearity range are analyzed, each determination
being repeated five times and the percentage relative standard deviation (% RSD) is
found to be less than 2 %. Accuracy of the proposed methods is measured by calculating
the percentage relative error (% RE) and is found to be less than 3 %. The results of this
study indicate the high accuracy and precision of the methods. Detailed results are given
in Tables 6.3A & 6.3B.
6.5.4.3. Selectivity
The effects of wide range of excipients which usually present in the formulations
are studied. It is found that proposed method can be successfully applied for the
determination of SFD in pharmaceutical formulation without any analytical problem due
to the tablet excipients such as glucose, starch and lactose.
The selectivity of the proposed methods is tested by preparing the placebo blank
(solution containing all the tablet excepients except SFD). A solution of analytical
placebo is prepared according to the sample preparation procedure and subjected to the
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analysis using the procedure described earlier. The absorbance measured is nearly same
as that of the reagent blank which indicates that the change in absorbance with respect to
the reagent blank caused only by the analyte. Non interference by common additives
including SFD is analyzed by preparing the test solution containing following
components: SFD (25 mg), glucose (45 mg), starch (50 mg) and lactose (30 mg). The
entire mixture is transferred to 100 mL calibrated flask, the contents shaken for 20 min;
volume diluted to the mark with distilled water, mixed and filtered. The filtrate after
suitable dilution is analyzed by proposed methods. The result confirms the selectivity of
the proposed methods in the presence of excipients.
6.5.4.4. Robustness
6.6. APPLICATIONS
The proposed methods are successfully applied for the determination of SFD in
dosage form. The results (Table 6.4) shows that the Student’s t tests at 95 % confidence
level are less than the theoretical values, which confirmed the good accuracy of the
methods. It is also clear that the result obtained from the proposed methods is in good
agreement with those obtained from the reported methods.
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6.7. CONCLUSIONS
❖ The proposed spectrophotometric methods are rapid and easily applicable for the
determination of SFD in tablets.
❖ The proposed methods are free from critical experimental conditions and
complicated procedures such as heating or extraction steps.
❖ The reagents used in the proposed methods are cheap, readily available and the
procedures do not involve any tedious sample preparation.
❖ These advantages encourage the application of the proposed methods in routine
quality control analysis of SFD in pharmaceutical formulation.
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Table 6.1: Comparison of A^of Schiff bases and Co(II) complexes synthesized from
different aryl aldehydes
Table 6.2: Spectral and statistical data for the determination of SFD
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Table 6.3A: Evaluation of accuracy and precision (Method A)
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o
u>
l/l
CO
d
CM
m
Absorbance
o o
CM
r-i
in
o o
Figure 6.1: Absorption maximum for (i) Co(II), (ii) method A and (iii) method B
CM
in
o
fN
Absorbance
Absorbance
Hm
O
oin
in
o
0 50 100 150
tH
Figure 6.2: Calibration curve for method A Figure 6.3: Calibration curve for method B
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Scheme 6.1: Reaction of PDAB and VA with SFD followed by the complex formation
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