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Sensors and Actuators B 127 (2007) 362369

Determination of paracetamol based on electropolymerized-molecularly imprinted polypyrrole modied pencil graphite electrode
Levent Ozcan, Y ucel S ahin
Anadolu University, Faculty of Science, Department of Chemistry, 26470 Eski sehir, Turkey Received 28 December 2006; received in revised form 21 March 2007; accepted 20 April 2007 Available online 4 May 2007

Abstract Preparation of a molecularly imprinted polymer (MIP) lm and its recognition property for paracetamol are investigated. The polypyrrole (PPy) lm was prepared by the cyclic voltammetric deposition of pyrrole (Py) in the presence of a supporting electrolyte (LiClO4 ) with and without a template molecule (paracetamol) through on a pencil graphite electrode (PGE). The performance of the imprinted and non-imprinted (NIP) lms was evaluated by differential pulse voltammetry (DPV). Several important parameters controlling the performance of the PPy was investigated and optimized. The molecularly imprinted lm exhibited a high selectivity and sensitivity toward paracetamol. The calibration curve for the DPV peak current observed for paracetamol oxidation versus paracetamol concentration at MIP electrode shows two linear regions. The rst region demonstrates linearity over a concentration range of 5 M to 0.50 mM with a correlation coefcient of 0.996. The slope of the second linear region was smaller than the rst regions slope with a wide concentration range of 1.254.5 mM (R2 = 0.990). The detection limit (3 ) of paracetamol is 7.9 107 M (S/N = 3). Molecularly imprinted polypyrrole modied pencil graphite electrode showed a stable and reproducible response without any inuence of interferents commonly existing in pharmaceutical samples. 2007 Elsevier B.V. All rights reserved.
Keywords: Molecularly imprinted polymer; Polypyrrole; Pencil graphite electrode; Electropolymerization; Paracetamol

1. Introduction Paracetamol (N-acetyl-p-aminophenol) is a commonly used analgesic and antipyretic drug these days [1]. Paracetamol (PC) was rstly introduced into medicine as an antipyretic/analgesic by Von Mering in 1893 and has been in use as an analgesic for home medication for over 30 years and is accepted as a very effective treatment for the relief of pain and fever in adults and children. It is the most used medicine after acetylsalicylic acid in many countries as an alternative to aspirin and phenacetin [2]. Over the past two decades, molecularly imprinted polymers (MIPs) have attracted broad interest from scientists engaged in sensor development. This attention can be explained by the serious potential advantages of using MIPs in place of natural receptors and enzymes such as their superior stability, low cost and easy preparation. The general principal of molecu-

Corresponding author. Tel.: +90 222 3350580; fax: +90 222 3204910. E-mail address: ysahin@anadolu.edu.tr (Y. S ahin).

lar imprinting is based on such a process where functional and cross-linking monomers are copolymerized in the presence of a target analyte (the imprint molecule) which acts as a molecular template. This procedure can be accomplished via either reversible covalent bonding or non-covalent interactions between monomers and imprint molecules. Other preparation methods of molecular imprinting polymers (MIPs) have been reported including chemical grafting, soft lithography technique [3], molecular self-assembled approach [4] and electropolymerization [3]. These lms can be synthesized in situ at an electrode surface by electropolymerization technique. This technique has some attractive features including the easy adherence of the polymeric lms to the surface of conducting electrodes of any shape and size and the ability to control thickness of the lms under different deposition conditions [5]. Various types of electrosynthesized polymers based on molecular imprinting have been reported in the literature including poly(ophenylenediamine) [5], polyphenol [6], polypyrrole [7] and copolymer of aniline with o-phenylenediamine [8]. Molecular imprinting has been approached using the electrosynthesis of conducting polymers through galvanostatic,

0925-4005/$ see front matter 2007 Elsevier B.V. All rights reserved. doi:10.1016/j.snb.2007.04.034

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potentiostatic and cyclic voltammetric methods. These methods provide a simple and rapid technique of controlling the thickness of the conductive polymer lm grown adherent to a transducer of any size and shape. The monomers that have been used in the design of molecularly imprinted conducting polymers include pyrrole, aniline and o-phenylenediamine. Imprinting of neutral compound-glucose has been achieved successfully by electropolymerization of o-phenylenediamine [5]. The big advantage of electropolymerization in comparison with other immobilization techniques lies in the ability to deposit a recognition lm at precise spot of the detector surface. It is possible to coat an electrode with complex geometry with homogeneous lm. The polymer thickness and deposition density is regulated by polymerization conditions (e.g., applied voltage). Deore and Freund reported a new approach for the electrosynthesis of saccharide-imprinted poly(aniline boronic acid) [9]. The method involves the formation of a saccharideaminophenylboronic acid complex in the presence of uoride to allow the electropolymerization of a self-doped, molecularly imprinted polyaniline. The formation of the anionic monomer complex enables electrochemical polymerization at near neutral pH (57) ensuring the incorporation of saccharide in the resulting, self-doped polymer. Molecularly imprinted polymers selective for uorescein, rhodamine or 2,4-dichlorophenoxyacetic acid (2,4-D) were electropolymerized onto graphite electrodes using an aqueous solution equimolar in resorsinol/ortho-phenylenediamine and in the presence of the template molecule. For the dyes, the MIP-coated electrodes showed higher afnity for their template molecule than for a non-template dye [10]. The caffeine-imprinted polymer was synthesized using galvanostatic electropolymerization of pyrrole monomer directly onto one of the gold electrodes of a 9 MHz AT-cut quartz crystal in the presence of caffeine. Caffeine molecules were entrapped in the matrix of polymer lm, and were removed by subsequent washing with water, leaving behind pores capable of recognizing the target analyte molecule [11]. A molecularly imprinted polymer was prepared by electropolymerization of pyrrole onto a stainless steel frit, using ochratoxin A as the template, in order to make a micro solid phase preconcentration device [12]. The use of a differential pulse voltammetry to determine the paracetamol using pencil graphite electrode prepared by imprinting electropolymerization was reported for the rst time in this work. Paracetamol was chosen as template molecule because of its popularity and electroactivity. Its successful application to the determination of paracetamol in commercial pharmaceutical tablets and syrup has been demonstrated. 2. Experimental 2.1. Chemicals and reagents Paracetamol (98%), dopamine (98.5%), phenacetin (98%), l-ascorbic acid (>99.5%), d-glucose (>99.5%), phenol (>99.5%), potassium chloride (99%) were obtained from Fluka (Steinheim, Germany). Potassium dihydrogen phosphate (>99.5%, Merck, Darmstat, Germany), potassium hydrogen

phosphate (98.5%, SigmaAldrich, Steinheim, Germany), lithium perchlorate (98%, Lancaster, Morecambe, England) and other reagents commercially available as analytical grade and used without further purication. Pyrrole (98%, Aldrich, Steinheim, Germany) was distilled repeatedly under vacuum until a colorless liquid was obtained and kept under nitrogen in darkness at 4 C. Stock solutions of paracetamol and buffer solutions were prepared by using ultra-pure deionized water (Sartorius). Minoset and MinosetPlus (Roche, Kocaeli, Turkey), Vermidon ( Ilsan Iltas , Kocaeli, Turkey), TylolHot (Nobel, D uzce, Turkey) paracetamol tablets and Calpol syrup (GlaxoSmithKline, Abdi Ibrahim Ilac A.S ., Istanbul, Turkey) were purchased from local pharmacy. Freshly prepared solutions of paracetamol were prepared each day owing to its low stability. 2.2. Apparatus Electrochemical studies were performed using Autolab PGSTAT 100 potentiostat-galvanostat controlled by a GPES 4.9 software (Ecochemie, The Netherlands). Three electrode system was used for all measurements; a pencil graphite electrode (PGE) as the working electrode and a Pt auxiliary electrode. All measurements carried out with an Ag/AgCl reference electrode. IONcheck45 model (Radiometer, France) pH-Ion meter was employed for pH measurements. Chromatographic measurements were carried out using a Agilent 1100 Series HPLC (Germany) consisting of a gradient pump, a C18 separator column (ACE , 5 m, 150 4.6 mm, Scotland) coupled with an UVVis detector and computer. The eluent used was methanol:water (80:20) mixture at a ow rate of 0.8 mL min1 . All separation was carried out at 25 C. The detection was performed at 254 nm for the compounds. 2.3. Preparation of MIP and NIP electrodes A Noki pencil model 2000 (Japan) was used as a holder for graphite leads (Tombo, HB, 0.5 mm diameter, Japan). Electrical contact with the lead was obtained by soldering a metallic wire to the metallic part. PGEs were washed with water and acetonitrile to remove the impurity and dried at room temperature before the experiments. Then, PGE was immersed the polymerization solution. The MIP was obtained by electrodeposition on the surface of the PGE using cyclic voltammetry in the potential range between 0.60 and +0.80 V during four cycles (scan rate: 100 mV/s) in aqueous solution of 0.1 M LiClO4 , 0.05 M pyrrole and 0.020 M paracetamol. After the electropolymerization process, the embedded paracetamol were then extracted to give a surface complimentary in shape and functionality to the original template paracetamol. Then the imprinted polymer was conditioned in 0.1 M KCl + 0.05 M phosphate buffer solution using potential cycling between 0.6 and +1.00 V. This process provided to remove the template inside the polymer. We determined the excess paracetamol in this solution using DPV by uncovered PGE (the data were not given here). A control electrode (non-imprinted polymer modied electrode, NIP) was

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prepared in every case under the same experimental condutions but without adding the paracetamol, to check the reliability of the measurements. 2.4. Electroanalytical measurements Differential pulse voltammetric measurements were carried out in a three-electrode cell, in 0.1 M KCl + 0.05 M phosphate buffer at pH 7.0. Before the measurements, electrolytic solutions were purged with nitrogen for 5 min. Current measurements were performed using differential pulse voltammetry (DPV) in the potential range between 0.00 and 0.80 V. To record differential pulse voltammograms, the following instrumental parameters were used: step potential 8 mV, modulation amplitude 50 mV; scan rate 15 mV/s. All electroanalytical measurement were made at room temperature. 2.5. Sample preparation Various commercial pharmaceutical tablets and syrup having paracetamol were examined for estimation of paracetamol. The tablets were nely powdered and dissolved in water. The solution was sonicated for 10 min and ltered. An aliquot of appropriate volume of stock solution was transferred into 250 mL volumetric ask and volume was completed with buffer (pH: 7.0). The syrup was transferred to a 100 mL ask and volume was adjusted to the same pH value with buffer solution. The samples were then spiked with appropriate amount of paracetamol for experiments. 2.6. The structure of interferents To investigate the selectivity of the MIP and NIP electrode in this work was evaluated in the presence of different interfering molecules. The structure of paracetamol and some possible interfering substances, dopamine, phenacetin, ascorbic acid, phenol and d-glucose were given in the Fig. 1.

3. Results and discussion 3.1. Electropolymerization of molecularly imprinted polypyrrole The electrochemical behavior of pyrrole was investigated in aqueous solution of 0.1 M LiClO4 using potential cycling between 0.6 and +1.40 V (versus Ag/AgCl). Electrooxidation of the pyrrole monomer occurs at the anode, and the resulting polymer deposits onto the surface of pencil graphite electrode. An anodic peak of pyrrole was observed at a peak potential of 1.10 V. The corresponding reduction process was not observed on the cyclic voltammogram. The oxidation peak corresponds to the formation of pyrrole radical cations. Fig. 2a demonstrates four cycles obtained in the same solution. The formation and growth of the polymer lm can be easily seen in this gure. The peaks due to the oxidation and reduction of the lm increase in intensity as the lm grows. A broad oxidation peak was observed at the peak potential of +0.10 V and reverse cathodic peak was seen at a peak potential of 0.20 V. For imprinted electropolymerizations, paracetamol was added to the electrochemical cell at a concentration of 20 mM. Fig. 2b demonstrated cyclic scans of electropolymerization of pyrrole in the presence of paracetamol. The effect of paracetamol (template) on the electropolymerization of pyrrole can be seen easily in this gure. The oxidation peak potential of polypyrrole shifted to more anodic potentials, from 0.1 to 0.30 V,

Fig. 1. The structure of paracetamol, dopamine, phenacetin, ascorbic acid, phenol and d-glucose.

Fig. 2. Cyclic voltammograms taken during the electropolymerization of pyrrole (0.05 M). (a) Multisweep cyclic voltammograms without and (b) with paracetamol (20 mM) onto a pencil graphite electrode (scan rate: 100 mV s1 ; supporting electrolyte: 0.1 M LiClO4 ; number of scans: 4).

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Fig. 4. Effect of pH on the MIP modied pencil graphite electrode response for 0.25 mM paracetamol in a phosphate buffer solution with KCl.

Fig. 3. Schematic representation of (a) imprinting and (b) removal of paracetamol from paracetamol imprinted polypyrrole modied pencil graphite electrode.

in the presence of paracetamol. This oxidation peak indicates that the template is becoming part of the polymeric chain [13]. Because of the polymerization solution was not stirred, the mass transfer was occurred by diffusion controlled process. Paracetamol molecules diffuses towards the surface of the PGE during the electropolymerization process and trapped into the polymer matrix. The creation of the molecular imprints favored by the diffusion of the electroactive template, generating a far higher number of recognition sites than those previously obtained with a non-electroactive template. If the template is non-electroactive, molecular diffusion towards the electrode is interrupted after the rst scans by the formation of the non-conductive polymeric layer, which prevents the template forming part of this layer and thereby reducing the number of recognition sites. During the electrodeposition of the conductive polymer, paracetamol template molecules are trapped in the polymer matrix as a result of the ability of these molecules to interact with the pyrrole units. The schematic representation of imprinting and removal of paracetamol from paracetamol imprinted polypyrrole modied pencil graphite electrode was shown in Fig. 3. The oxygen atom in the C O group of the paracetamol molecule forms a hydrogen bond with the hydrogen atom in the NH group of the pyrrole units (Fig. 3). Hydrogen bonding could

occur between the hydrogen in the hydroxyl group of paracetamol structure and the nitrogen atom of the NH group of pyrrole units. There may be a hydrogen bond between nitrogen atom in the NH group of the paracetamol molecule and the hydrogen atom of the NH group of pyrrole units. Chain branching and cross linking in PPy [14] generate a three-dimensional matrix with niches containing the template paracetamol. This imprinting process creates a microenvironment for the recognition of paracetamol molecule based on shape selection and positioning of the functional groups. In order to remove the entrapped template, various methods are possible: microwave assisted extraction, oxidation-reduction of the template in the polymer or the use of solvent that strongly interacts with the polymer causing the swelling of the coating necessary for template release. To remove the template inside the polypyrrole, the imprinted polymer was conditioned in 0.1 M KCl + 0.05 M phosphate buffer solution using potential cycling between 0.6 and +1.00 V in our work. 3.2. Effect of pH The pH of the solution has a signicant inuence on the stability of the polymeric lm. The signal of the polymer should be the lowest to avoid an interaction with paracetamol signal. It can be concluded that the polymer response was the lowest at neutral pH, while at more acidic and basic pH there is a corresponding signal to the polymer structure. Therefore the pH of the medium in which the measurement with the polypyrrole modied pencil graphite electrode is to be made should be equal to 7.0. The mechanism for the electrochemical oxidation of paracetamol is different depending on the pH of the medium. The pH effect of the MIP electrode is illustrated in Fig. 4 for paracetamol (0.25 mM) at different pH, in the range of 311, by DPV. The paracetamol signal shifted to more cathodic potentials as the pH increases (Fig. 4). The best results are obtained at a pH of 7.0, with a 0.05 M phosphate and 0.1 M KCl, giving an oxidation peak at 450 mV. No further improvements were observed at higher ionic strength.

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Fig. 5. Differential pulse voltammograms for (a) 0.25 mM and (b) 1.0 mM paracetamol at non-imprinted polypyrrole (NIP) (c) 0.25 mM and (d) 1.0 mM paracetamol at molecularly imprinted (MIP) modied pencil graphite electrode in a phosphate buffer medium at pH 7.0 with 0.1 M KCl.

3.3. Electrochemical behavior of paracetamol The electrochemical behavior of paracetamol was investigated by MIP and NIP electrode. The catalytic effect of the MIP electrode is demonstrated in Fig. 5 for paracetamol (0.25 mM) at pH 7.0 recorded at a NIP electrode and the MIP electrode by DPV. Paracetamol gives an oxidation peak response at about 370 mV versus at the NIP electrode, while the use of MIP electrode leads to an anodic peak at about 450 mV versus Ag/AgCl and the peak current increased greatly. In neutral media the paracetamol oxidizes to N-acetyl-p-quinoneimine [13]. The enhanced peak current response and a shift in the oxidation potential of paracetamol by about 80 mV in the anodic direction are a clear evidence of the catalytic effect of the MIP electrode towards the oxidation of paracetamol. The calibration curves were observed for paracetamol at MIP and NIP electrode at pH 7.0. The calibration curve for the DPV peak current observed for paracetamol oxidation versus paracetamol concentration at MIP electrode shows two linear regions. The rst region demonstrates linearity over a concentration range of 5 M to 0.50 mM with a correlation coefcient of 0.996. The slope of the second linear region was smaller than the rst regions slope with a wide concentration range of 1.254.5 mM (0.990). The detection limit (3 ) of paracetamol is 7.9 107 M (S/N = 3). The percentage error in the calculated detection limit was estimated to be 1.0%. Modication of PPy surface by imprinting remarkably improves the reactivity of MIP electrode towards the oxidation of paracetamol, thus, making it possible to detect paracetamol in a solution up to as low as 0.5 M. The calibration curve observed for paracetamol at NIP electrode was also show a linearity over a concentration range of 50 M to 0.50 mM. However, the current range was much smaller than that of observed at MIP electrode. The paracetamol oxidation current remained constant between 2.0 and 5 mM. 3.4. Effect of the monomer concentration The monomer concentration during polymerization also determined the analytical behavior of the sensor. Electropoly-

Fig. 6. Effect of the monomer concentration on ( ) molecularly imprinted (MIP) and ( ) non-imprinted polypyrrole (NIP) modied pencil graphite electrode for 0.25 mM paracetamol. Response was measured thorough DPV in 0.25 mM paracetamol in a phosphate buffer medium at pH 7.0 with 0.1 M KCl.

merization using low monomer concentrations produced sensors with good analytical behavior. To determine the effect of monomer concentration on the response of both MIP and NIP to paracetamol, the lms were grown in solutions of constant concentration of paracetamol and varying pyrrole concentrations in the range of 25500 mM by cycling potential between 0.60 V and +0.80 V for the same cycles. Fig. 6 shows the results of such a comparison. The binding ability of the template molecules onto the coating lm depends on the number of imprints. The monomer concentration should be proportional to the thickness of the deposit and amount of imprinted molecule (template) in the polymeric matrix. The response of the MIP electrode to paracetamol was found to increase with increasing pyrrole concentration up to 50 mM. There was considerable decrease in the response of MIP electrode below and above this pyrrole concentration. The current difference between the MIP and NIP electrode for paracetamol should be as high as possible (iMIP iNIP ). In addition, the signal of the NIP electrode should be nearby zero (iNIP 0). It can be concluded that the optimum monomer concentration under these conditions was about 50 mM. 3.5. Effect of the electropolymerization cycles The optimum number of CV cycles to use to form the sensing layer of the electrode was determined from a series of experiments in which electrodes were fabricated with different numbers of cycles in aqueous solution of 0.1 M LiClO4 , 0.05 M Py and 20 mM paracetamol by cycling potential between 0.60 and +0.80 V. The number of cycles applied to the cell during the electropolymerization was found to affect the sensitivity and linearity of the sensor. The MIP electrode produced at lower number of cycles exhibited favorable analytical performance. Higher cycles lead to more extensive electropolymerization, and therefore to the formation of thicker sensing lm with less accessible imprinted sites. The response of the MIP and NIP electrodes to paracetamol was found to increase with increasing the number of cycles up to 4 and 6, respectively (Fig. 7). There

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Fig. 7. Effect of the number of cycle to ( ) molecularly imprinted (MIP) and ( ) non-imprinted polypyrrole (NIP) modied pencil graphite electrode response in a phosphate buffer medium at pH 7.0 with 0.1 M KCl.

was considerable decrease in the performance of the MIP electrode below and above this number of cycles. The highest current difference between the MIP and NIP electrode for paracetamol was obtained by applying 4 cycles in the electropolymerization. Therefore the optimum polymerization cycles was found to be 4. The thickness of the PPy was measured by the charge passed through the electropolymerization process [15] was about 500 nm. 3.6. Effect of the template concentration Dopamine is a most signicant neurotransmitter, plays a prime role in the functioning of the central nervous, cardiovascular and hormonal systems and the structure of dopamine is quite similar to paracetamol. Dopamine is present in biological uids and may interfere with paracetamol because of its similarity to that of paracetamol and electrochemically active. The structures of dopamine and the other interferents are given in Fig. 1. The oxidation peak potentials are +0.18 and +0.45 V for dopamine and paracetamol, respectively at MIP electrode. 0.27 V difference in the oxidation peak potentials of dopamine and paracetamol was allowed to analyse paracetamol without any interaction with dopamine in biological samples. The effect of the template concentration during the lm electrodeposition was shown in Fig. 8. Dopamine was used as a control substance to investigate the effect of template (paracetamol) concentration on the response of the MIP electrode in this gure.The sensor behavior was inuenced by the template concentration employed in the electrodeposition process. As the template concentration grows in the polymerization solution, the response to paracetamol of the MIP electrode obtained is also higher. The great number of specic cavities for paracetamol was generated using high concentration of template. However, this process may cause decrease the selectivity of the sensor. Because of this limitation, a greater number of cavities will permit a greater diffusion of paracetamol to the active surface of the MIP electrode. The response of the MIP electrode to paracetamol was found to increase with increasing paracetamol concentration up to 5 and 20 mM (Fig. 8). There was a small decrease in the response of MIP electrode above this paracetamol concentration (20 mM).

Fig. 8. Response of MIP electrode was measured through DPV in solutions with ( ) paracetamol as a template and ( ) dopamine as a control substance to MIP electrode response in a phosphate buffer medium at pH 7.0 with 0.1 M KCl.

It is necessary to select a template concentration to which the response to paracetamol is highest, without obtaining a signal for dopamine. The highest current difference between paracetamol and dopamine was obtained with 20 mM paracetamol concentration. Therefore the optimum template concentration was found to be 20 mM. 3.7. Effect of interferents The selectivity of the MIP and NIP electrode in this work was evaluated in presence of different interfering molecules. Voltammetric responses of paracetamol imprinted and nonimprinted polypyrrole lm were examined in the presence of some possible interfering substances like ascorbic acid, dglucose, phenacetin, dopamine and phenol. These substances are present in biological uids and may interfere with the determination of paracetamol through conventional methods. Differential pulse voltammograms were taken for the oxidation of paracetamol (0.25 mM) after addition of varying concentration of each interferent (0.252.5 mM). The obtained currents in the absence of any interferent were 42 and 115 A with NIP and MIP electrode, respectively. The results are given in Table 1. In all cases, there was no substantial change in current response for 0.25 mM paracetamol in the presence of less than two fold excess of interferents for MIP electrode. At higher concentrations of these interferents the variation was within 7.170 A relative to that in their absence. However, the concentrations of phenacetin and dopamine are not as high as 1.0 mM in biological samples. Thus, the response of paracetamol at the MIP electrode is not affected by the interferents examined here below at least four fold concentrations. It was found that D-glucose and phenol produce negligible changes in the paracetamol response even when an analyte-inteferent ratio of 1:10 was used. That is, the electropolymerized-molecularly imprinted polypyrrole modied pencil graphite electrode can recognize the paracetamol molecules by means of shape selection and the size of functional groups. However, the response of paracetamol at the NIP electrode is affected by the interferents.

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Table 1 Effect of interferents on the differential pulse voltammetric response of 0.25 mM paracetamol at the MIP and NIP electrode Interferent Phenol Concentration of interferents (mM) 0.25 1.00 2.50 0.25 1.00 2.50 0.25 0.50 1.00 0.25 0.50 1.00 0.25 0.50 1.00 Change in current responsea for 0.25 mM paracetamol with MIP (A) 0.678 1.150 2.100 0.852 1.495 2.645 2.163 3.245 7.170 1.370 2.980 5.250 0.62 1.78 2.350 Change in current responseb for 0.25 mM paracetamol with NIP (A) 3.360 7.351 10.990 0.823 3.884 5.209 8.863 12.518 19.021 6.813 12.930 15.280 4.691 7.195 10.033

Glucose

Phenacetin

Dopamine

Ascorbic acid

a b

The current in the absence of any interferent was 115 A (with MIP). The current in the absence of any interferent was 42 A (with NIP).

Table 2 A comparison of observed and reported paracetamol concentration in pharmaceutical preparations (tablets and syrups) using DPV at molecularly imprinted polypyrrole modied pencil graphite electrode and HPLC Tablet/syrup name Minoset Minoset Plus Vermidon TylolHot Calpol syrup
a b c

Reported content 0.500a 0.250a 0.500a 0.500b 120c

Detected content (with MIP) 0.487a 0.236a 0.496a 0.492b 117c

R.S.D. (%) (n = 3) 1.26 0.67 0.81 1.62 1.55

Detected content (with HPLC) 0.496a 0.245a 0.495a 0.494b 118c

R.S.D. (%) (n = 3) 0.42 0.49 0.55 0.48 0.31

g/tablet. g/20 g. mg/5 mL.

3.8. Analysis of commercial samples Various tablets and syrup having paracetamol were examined for estimation of paracetamol by MIP electrode and HPLC method. Solution obtained by dissolution of paracetamol tablets and syrup were subsequently diluted so that paracetamol concentration lies in the range of calibration plot. Differential pulse voltammograms were then recorded under exactly identical conditions that were employed while recording differential pulse voltammograms for plotting calibration plot. Keeping dilution factor in consideration, it was found that paracetamol concentration determined using this method is in good agreement with the manufacturers stated contents of paracetamol (as shown in Table 2). Experimentally determined paracetamol amount and reported paracetamol amount in tablets and syrup are listed in Table 2. In order to validate the electrochemical detection we have compared the obtained results with both detection methods, by using the optimal chromatographic conditions. Table 2 shows the obtained data when the HPLC and DPV, using MIP electrode, are used. In this table we can observe that both detection methods were useful for analytical purposes.

3.9. Reproducibility of the MIP electrode The reproducibility of the molecularly imprinted modied pencil graphite electrode was investigated for 0.25 mM paracetamol. The peak current response of paracetamol was determined with six electrodes which produced under the same conditions. The response peak intensity showed a relative standard deviation of 1.3% conrming that the results are reproducible. The MIP PG electrodes could be used at least 10 or 15 times with subsequent washing and measuring operations. Alternatively another MIP-PGE could be easily prepared with the procedure described above. 4. Conclusion Electrochemistry has many advantages making it an appealing choice for pharmaceutical analysis. The role of electrochemistry in pharmaceutical analysis has been well dened, and is likely to get preference when low analyte concentrations, small sample volumes, or complex sample matrices requiring high specicity challenge the analytical method. In this study, paracetamol imprinted electrodes formed by the cyclic

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voltammetric deposition of polypyrrole lm on pencil graphite electrode in the presence of target paracetamol have been successfully fabricated. Molecular imprinting was shown to be an effective way of growing the polymer coating directly onto a disposable electrode. A linear relationship between paracetamol concentration and current response was obtained with excellent reproducibility of the current and a low detection limit of 7.9 107 M. Molecularly imprinted polypyrrole modied pencil graphite electrode showed a stable and reproducible response without any inuence of interferents commonly existing in pharmaceutical samples. The proposed low cost chemical sensor could nd application in the measurement of paracetamol level in clinical samples as well as in pharmaceutical industry. Acknowledgements Financial support of Anadolu University Research Found (Project Nos.: 031064 and 051060) is gratefully acknowledged. References
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Biographies
Levent Ozcan was born in Sakarya, Turkey on October 16, 1977. He received his bachelor degree in chemistry from Anadolu University, Eskis ehir, Turkey in 1999. In 2002 he has received his master degree in Department of Chemistry at Anadolu University. He has been a PhD student in Department of Chemistry at Anadolu University since 2002. He research interests include electrochemical biosensors, modied electrodes and conducting polymers. Yucel S ahin was born in Ankara, Turkey in 1969. He received his master and PhD degrees in the Department of Chemistry at Hacettepe University in 1996 and 2000, respectively. He has worked as an assistant professor from 2000 to 2004 in the Department of Chemistry at Anadolu University. Since 2004, he worked as an associated professor in the same department. His current research interests are electroanalytical chemistry, conducting polymers, electrochemical sensors and biosensors, modied electrodes, electrocatalysis, and electroorganic chemistry.