Journal of the Chinese Chemical Society, 2004, 51, 363-366

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A New Kinetic-Photometric Method for Determination of Carbimazole

Mohsen Barzegara*, Abdolvahed Rahmanib, Ali Jabbaric and Mir Fazlollah Mousavid a Department of Food Sciences and Technology, Tarbiat Modarres University, P. O. Box: 14115-336, Tehran, Iran b Department of Chemistry, Hormozgan University, Bandar Abbas, Iran c Department of Chemistry, K. N. Toosi University, Tehran, Iran d Department of Chemistry, Tarbiat Modarres University, Tehran, Iran

A new kinetic photometric method for determination of carbimazole (CBZ) is described. The proposed method is simple, rapid, inexpensive and sensitive for the determination of CBZ in pure and tablet forms. This method is based on the inhibitory effect of carbimazole on the palladium(II)-catalyzed reaction between neutral red (NR) and hypophosphite (HP) ions. The effect of various parameters such as dye, hypophosphite, and Pd(II) concentrations, pH, ionic strength, and temperature were optimized. The calibration graph was linear in the range 0.04-0.30 ppm (n = 9, r = 0.9972) and the detection limit was 0.02 ppm. The relative standard deviation of the developed method was 0.70% (n = 10). The proposed method was applied for the determination of carbimazole in pure and tablet forms.
Keywords: Carbimazole; Kinetic photometric method; Palladium; Inhibition; Neutral red.

INTRODUCTION Carbimazole 3-ethoxycarbonyl-1-methyl-2,3-dihydro1H-imidazole-2-thione (Scheme I) is an anti-thyroid agent that depresses the formation of thyroid hormones.1 Scheme I Carbimazole

N Me S

N COOEt

fluorometric.15 Some of the published methods suffer interference from tablet base, while others are not simple for routine analysis, as they need sophisticated instruments and expensive reagents. Therefore, it was considered worthwhile to develop a rapid, simple and accurate procedure suitable for application in quality control laboratories. In this paper, a detailed study of the appropriate conditions for the inhibition effect of CBZ on the reduction of neutral red (NR) by hypophosphite (HP) in the presence of palladium(II) was performed. The reaction was monitored photometrically at the maximum wavelength of the NR (530 nm) while measuring the change in the absorbance with time.

This drug and other anti-thyroid agents being the thiourea pharmacophore, inhibit the first step in the pathway of thyroid hormone biosynthesis which is the incorporation of oxidized iodide into tyrosine residues in the large thyroid hormone precursor molecule, thyroglobulin. The determination of carbimazole (CBZ) is important in different areas such as clinical chemistry, nutrition and pharmaceutical formulations. Several analytical procedures have been described for the analysis of CBZ: an indirect bromometric titration,2 a direct titration method,3 potentiometric,4,5 voltammetric,6 chromatographic,7-9 spectrophotometric,10-13 polarographic,14 and
* Corresponding author. E-mail: mbb@modares.ac.ir

EXPERIMENTAL Reagents All chemicals were of analytical-reagent grade and the solutions were prepared with doubly distilled water. Working solutions of lower concentrations were prepared by appropriate dilution of the respective stock solutions prior to use. Palladium chloride (1000 ppm, 0.1670 g) was dissolved in 2.0 mL concentrated HCl (Merck) and the solution was transferred into a 100 mL standard flask and then diluted to

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volume with water to make Pd stock solution. Neutral red solution (2 10-4 M) was prepared by dissolving 0.0290 g of NR (Merck) and diluting to 500 mL in a calibrated flask. Hypophosphite (2 M) solution was prepared by dissolving 20.8180 g of potassium hypophosphite (Merck) in 100 mL of water. The purity of the carbimazole, kindly supplied by Poursina Pharmaceutical Company (Tehran, Iran), was proved to be 100.4% according to BP.16 The stock solution was prepared by dissolving 0.0100 g of CBZ with water and diluting to 100 mL. The pharmaceutical formulation carbimazole tablets (5 mg) were offered by Poursina Pharmaceutical Company. Apparatus A 662 probe-type photometer (Metrohm), and a Grant thermostatically controlled bath/circulator (Grant Instrument, Ltd., Cambridge) were used. During all experiments, solutions were stirred with a magnetic stirrer (Zag-Chemie Co., Ltd., Tehran, Iran). All pH measurements were carried out with a Metrohm pH-meter model 691. Procedures For calibration curve: After a suitable aliquot of solution containing 0.6-4.5 mg carbimazole was transferred into a 15-mL volumetric flask, 3.5 mL of 2.0 M hypophosphite, and 3.0 mL of 2.0 10-4 M neutral red solution were added. The solution was diluted to 15 mL with water. After mixing well, the aliquot was transferred into a glass cell (thermostated at 25 C). Then, the A-t curve was recorded just after the instantaneous addition of Pd(II) using a microsyringe. The reaction was monitored photometrically by measuring the absorbance at 530 nm. The time scan was recorded by computer-interfaced probe-type. The cell was cleaned after use by immersion into HNO3 (6 M) in order to remove any trace of Pd(0) absorbed on to the wall. Assay of CBZ tablet: Ten tablets were weighed and crushed in a mortar. A mass of powder equivalent to the average mass of one tablet (5 mg) was dissolved in 50 mL water. The solution was filtered through a Millipore filter, and the filtrate was diluted with water in a 100 mL calibrated flask. A 2.0 mL aliquot of solution was diluted to 50 mL with water and the proposed procedure was applied. All the necessary precautions were taken to guard against the possible degradation of CBZ. These precautions included the use of freshly prepared solution at low temperature and protection from light.

RESULTS AND DISCUSSION Our previous work17 has demonstrated that in the presence of a trace amount of Pd(II), hypophosphite reduces neutral red rapidly. A report11 has shown that Pd(II) reacts with CBZ and methimazole to produce yellow complexes. Therefore, carbimazole decreases the catalytic effect of Pd(II) on the reaction of the NR-HP system. The reaction rate of the catalyzed reaction decreased with increasing the amounts of carbimazole and time required for the absorbance to decrease to a predetermined value increased. The reaction was monitored photometrically by measuring the time required for the absorbance of the NR to decrease by 0.1 (at lmax = 530 nm) by means of variable time method. Effect of variables To obtain the maximum sensitivity in the determination of CBZ, we investigated the effect of several variables on the rate of catalyzed and inhibited catalyzed reactions, as follows. The effect of pH on the catalyzed and inhibited catalyzed reaction was studied in the range of 2.0-6.0 and the results are shown in Fig. 1. As can be seen, the inhibitory effect of CBZ reaches a maximum and is constant over the pH range 4.1-5.0. All subsequent investigations were performed at pH = 4.6. The influence of the hypophosphite concentration was studied over the range 0.11-0.56 M. The rates of catalyzed and inhibited reactions increase with increasing concentration of HP up to 0.39 M, above which they decrease slightly. The effect of neutral red was investigated in the range 1.1 10-5-5.6 10-5 M. Fig. 2 shows the percentage of inhibition (I%) versus different NR concentrations. A concentration of 3.3 10-5 M, at which the inhibitory effect of CBZ is
25 20

%I

15 10 5 0 1.5 2.5 3.5 4.5 5.5 6.5

pH

Fig. 1. Effect of pH on the percent of inhibition. Conditions: [NR] = 3.3 10-5 M; [Pd] = 1.1 ppm; [HP] = 0.39 M; T = 25 C.

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maximum, was selected. The influence of ionic strength by using KCl solution (2 M) on the reaction rate was studied in the concentration range of 0.0-0.9 M. The percentage of inhibition remains constant when the KCl concentration is in the range of 0.0-0.34 M and decreases with the concentration larger than 0.34 M. An important variable is concentration of Pd(II). The inhibitory effect of the antithyroid drug decreases when the concentration of Pd(II) increases; see Fig. 3, which shows the results obtained in the presence of 0.14 ppm of CBZ and an increasing amount of Pd(II). Moreover, the linear range of CBZ concentrations that can be determined and the slope of the calibration graph obtained under optimum conditions depend on the concentration of catalyst. A concentration of 0.45 ppm of Pd(II) was selected as the most suitable concentration. The effect of temperature on the rate of catalyzed and inhibited reaction was studied in the range 15-55 C. The results show that 25 C is best, since at higher temperatures the inhibition effect of CBZ is decreased, causing great increase in the rate. Thus, 25 C was used as the most convenient temperature.
25 20

Table 1. Determination of Carbimazole in Tablet Form in mg per Tableta Declared content 5

a

Reference method Proposed method RSD% 4.90 0.10 4.93 0.04 0.81

Average of six determinations S.D.

Features of the analytical method The method used to construct calibration plot in the kinetic determination of inhibitors, as described by PerezBendito,18 was commonly run by plotting the percentage of inhibition as a function of the inhibitor concentration. Under the optimum experimental conditions a calibration plot was obtained in the range 0.04-0.30 ppm of carbimazole. In the concentration range 0.04-0.30 ppm of CBZ, a regression equation, I% = 40.58 + 0.20 C, with a correlation coefficient of 0.9972 (n = 9) was obtained, where C is the concentration of carbimazole (ppm). The theoretical limit of detection19 was 0.02 ppm of CBZ. The precision (RSD%) of the developed method was 0.70% (n = 10) for 0.15 ppm of carbimazole. Study on the interference from other substances Under the optimum experimental conditions to study the selectivity of the method, the effect of frequently encountered excipients and additives in the pharmaceutical dosage form of CBZ was studied by adding different amounts of possible interferences to a sample containing 0.17 ppm of CBZ. No interference was observed from the presence of glucose, lactose, starch or carboxymethyl cellulose (120-fold).

%I

15 10 5 0 0 1 2 3 4
5

C (M) x 10

Fig. 2. Influence of neutral red concentration on the percent of inhibition. Conditions: [Pd] = 1.1 ppm; [HP] = 0.39 M; T = 25 C.
80 60

40 20 0 0 0.5 1 C (ppm ) 1.5 2

Application To evaluate the analytical applicability of the method, the developed method was applied to the determination of CBZ in carbimazole tablets. For comparison purposes, the reference method adopted in the BP16 was applied using spectrophotometric determination at 291 nm. Table 1 shows the results obtained by proposed and reference methods. Applying the F-test and the t-test at the 95% confidence level compared the results obtained in both methods. The calculated F and t values did not exceed the theoretical values (F = 5.05, t = 2.57) indicating that there is no significant difference between the two methods with respect to accuracy.

Fig. 3. Effect of Pd(II) concentration on the percent of inhibition. Conditions: [NR] = 3.3 10 -5 M; [HP] = 0.39 M; T = 25 C.

%I

CONCLUSION The kinetic-photometric method proposed for the de-

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termination of CBZ is simple, sensitive, inexpensive, and rapid. This method is comparable in accuracy and precision with the reference method described in the cited reference. The developed method is suitable for the analysis of the CBZ in pharmaceuticals, as there are no interferences from the excipients normally found in commercial preparations.

ACKNOWLEDGEMENT The authors are grateful to the Tarbiat Modarres University Research Council for financial support and Poursina Pharmaceutical Company for supplying pure and tablet forms of carbimazole.

Received May 27, 2003.

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