International Journal of Biomedical and Advance Research

VALIDATED UV SPECTROSCOPIC METHOD FOR ESTIMATION OF ZOLMITRIPTAN FROM TABLET FORMULATIONS Syeda Humaira, N.G.Raghavendra Rao*, Mangesh R. Munde PG Department, Luqman College of Pharmacy, Old Jewargi Road, Gulbarga-585102, Karnataka, India. E-mail of Corresponding author: ngraghu@rediffmail.com

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Abstract A simple, economical, sensitive and specific UV spectrophotometric method was developed for the estimation of Zolmitriptan in bulk drug tablet dosage form. The optimum conditions for the analysis of the drug were established. The wavelength maxima (max) for zolmitriptan were found to be 226.5 nm. The linearity for this method was found to be in the range of 1-5g/ml. The method showed high sensitivity with reproducibility in results. The lower limit of detection and the limit of quantification were found to be 4.234and 12.702 respectively. The calibration curve was drawn by plotting graph between absorbance and concentration. Coefficient of correlation was 0.99. The regression of the curve was Y = 0.1751x +0.0476. Precision of the method was found to be 1.625 0.324 against the label claim of 2.5mg. The percentage recovery was found to be 98.66 99.50. The sample solution was stable up to 12 hours. The proposed method may be suitably applied for the analysis of Zolmitriptan in tablet pharmaceutical formulation for routine analysis Keywords: Zolmitriptan, UV Spectroscopy, tablet dosage form. Introduction Zolmitriptan[1,2], single enantiomer (4S)-4[[3-[2-(dimethylamino)ethyl]-1H-indole-5yl]methyl]-2-oxazolidinone is a novel serotonin 5-receptor agonist that has shown ,in an extensive clinical trial program, to be highly effective in the acute oral treatment of migraine with or without aura [3,4]. It works by stimulating serotonin receptors in the brain serotonin is a natural substance in the brain that, among other things, causes blood vessels in the brain to narrow. Zolmitriptan mimics this action of serotonin by directly stimulating the serotonin receptors in the brain and relieves the pain of migraines.

Chemical structure of zolmitriptan A few techniques had been reported in the literature for zolmitriptan quantitation in plasma samples coulometric detection, liquid chromatography tandem mass spectroscopy, liquid chromatography with florescence detection and chiral liquid chromatographic method for drug determination and its impurities [5-8]. www.ijbar.ssjournals.com

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Further a Spectrophotometric method is also reported in the literature [9]. To the best of our knowledge there are no UV Spectrophotometric methods reported in the literature. This paper describes for the first time the application of UV spectroscopy for the quantitative analysis of zolmitriptan in pharmaceutical raw material and dosage forms. Materials and Methods Instruments: UV-Visible double beam spectrophotometer (UV-1700, SHIMADZU Co, Japan) with 1 cm matched quartz cells and digital balance (Citizen Co.) were used. Materials: Zolmitriptan was obtained as gift sample from Cipla Pvt. Ltd, Mumbai. The obtained zolmitriptan was having 99.99% w/w assay value and was used without further purification. Ethanol was purchased from Changshu Yangyuan Chemicals, China. All chemicals and reagents used were of analytical grade. The zolmitriptan tablets were purchased from local market of Gulbarga. Preparation of standard stock solution: The standard stock solution was prepared by dissolving zolmitriptan in ethanol to make final concentration of 100 g/ml. Different aliquots were taken from stock solution and diluted with ethanol separately to prepare series of concentrations from 1-5 g/ml. The max was found by scanning UV spectrum of zolmitriptan in ethanol, in the range of 200-400 nm and it was found to be 226.5 nm. Absorbance was measured at 226.5 nm against ethanol as blank. The calibration curve was prepared by plotting absorbance versus concentration (g/ml) of zolmitriptan shown in Fig 1. IJBAR(2010)01(03)

Application of the proposed procedure for the determination in tablets: The proposed method was applied in order to determine the zolmitriptan in tablet formulation. The marketed tablet formulation of zolmitriptan was used for this. Twenty five tablets were weighed and average weight was calculated, crushed to fine powder. The powder equivalent to100 mg of zolmitriptan was transferred in 100 ml volumetric flask and dissolved in ethanol by shaking. The solution was then filtered through whattman filter paper # 41. The volume was made up to mark to get final concentration of 1mg/ml.The above solution was diluted suitably with ethanol to get the solution of 100g/ml concentration. The working solution of drug (100g/ml) was prepared from standard stock solution in ethanol. The absorbance of this solution was measured and amount of zolmitriptan was calculated from the calibration curve. The readings were taken in triplicate. Method Validation: As per the ICH guidelines, accuracy, precision, LOD, LOQ and linearity of the calibration curve were determined [10-12]. For linear response measurement, the least squares method was applied. The statistical analysis was calculated by ANOVA. Amounts of 1 g/ml,2 g/ml and 3 g/ml of zolmitriptan standard solution were added into pre analyzed samples and absorbances were measured and the recovery was calculated. Results and Discussion The development of spectrophotometric methods for the determination of drugs has increased considerably in recent years because of their importance in pharmaceutical analysis. Due to its greater solubility in ethanol, it was selected for further study. The values of standard www.ijbar.ssjournals.com

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deviation and coefficient of variation were satisfactorily low. The percentage recovery range of 98% to 101% was indicating the accuracy of method. From the proposed method, it was found that zolmitriptan obeys linearity within the concentration range of 1-5 g/ml shown in Fig 2. It was found that the % RSD is less than 2, which indicates that the method is highly reproducible. The analytical and necessary validation parameters for the UV spectrophotometric determination of Zolmitriptan from tablet formulation are presented in Table 1 For evaluating precision, the selected concentration (2.5 and 4g/ml) were prepared in ethanol and analyzed to determine the intraday and inter day variability. The intra-day and inter day precision were determined and presented as the RSD %. The precision and accuracy of the analysis are given in Tables 2, indicates high precision and accuracy. Determinations of active ingredients in tablets were analyzed and results are shown in Table 2. For evaluating precision, the selected concentration (2.5 and 5 g/ml) were prepared in ethanol and analyzed to determine the intraday and inter day variability. The intra-day and inter day precision were determined and presented as the RSD %. The Precision and accuracy of the analysis are given in Table 3, indicates high precision and accuracy. Recovery studies: The accuracy and validity of the proposed method were further ascertained by performing recovery studies. Pre-analysed tablet powder was spiked with pure zolmitriptan at three concentration levels(50.100,150% of that in tablet powder) and the total was found by proposed methods. In all cases, the added IJBAR(2010)01(03)

zolmitriptan recovery percentage values ranged between 98.66-99.50 (Table-4) indicating that the recovery was good. Conclusion: The proposed methods can be applied for quantitative assay of zolmitriptan in tablet dosage form. The method was applied on commercially available brand of zolmitriptan and results showed that the drug content of this formulation was in accordance with the labeled claim 2.5mg (Zomig, Astra Zeneca). Acknowledgement: The authors are thankful to Cipla Pvt. Ltd, Mumbai procuring Zolmitriptan as gift sample. Authors are also thankful to Prof. Syed Sanaullah, Principal Luqman College of Pharmacy for providing all facilities to complete this work successfully References 1. Sweetman SC. (Ed.), Martindale- the Complete Drug Reference Pharmaceutical Press, London (U. K.) 2002 33th edition., p.458. 2. Oneil MJ (Ed.), the Merck Index An Encyclopedia for Chemicals, Drugs and Biologicals., Merck and Co., 2001 13th edition., p. 1816. 3. Yates Y, Niarn K, Dixon R, Kemp JV, Dane AL. J. Clin. Pharmacol. (42), 2002 , 1244-1250. 4. Oldman AD, Smith LA, McQuary HJ, Moore RA, Pain 2002, 97, 247-257. 5. Clement EM, Franklin M, Chromatogr J. Analyst Technol.Biomed.Life Sci. 2002, 766, 339- 343. 6. Vishwanathan K, Barlett MG, Stewart JT. Rapid Commun. Mass Spectrum. 2000, 14, 168-172. www.ijbar.ssjournals.com

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7. Chen J, Jiang X, Jiang W, Mei N, Gao X, Zhang Q. J. Pharm. Biomed, Anal. 2004, 35, 639- 645. 8. Srinivasu MK, Mallikarun Rao B, Sridhar G, Rajendra Kumar P, Chandrasekhar KB, J. Pharm. Biomed, Anal.2005, 37, 453- 645. 9. Asad Raza, Tariq Mahmood Ansari, Shahida Begum Niazi, J. Chin. Chem. Soc. 2007, 54(6), 1413-1417. 10. International Conference on Harmonization (ICH), Harmonized Tripartite Guidelines on Validation of Analytical Procedures: Methodology,

Operational from June 1997, Published by The European Agency for the Evaluation of Medicinal products, Human Medicines Evaluation Unit. 11. International Conference on Harmonization, Draft Guideline on Validation Procedure, Definition and Terminology Federal Register, 1995, 60, 11260. 12. SO Krause, Supp to Bio Pharm Int. 2005, 2634.

Table 1: Analytical validation parameters:

Parameter Result

Absorption maxima(nm) Linearity Range (g/ml) Molar Absorptivity Regression equation (Y*) Slope (b) Intercept(a) Correlation Coefficient (r2 ) Accuracy (% recovery SD) Precision (%) LOD (g/ml) LOQ (g/ml) %Drug found in tablet Formulation Y=(bC+a)*

226.5 nm 1-5 7.7 X104 0.1751 0.0476 0.9987 98.56 0.238 1.625 0.324(Intraday) 1.279 0.215(Interday) 4.234 12.702 99.53

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Table 2: Determinations of active ingredients in tablets Sample Zolmitriptan Label claimed 2.5 mg Amount Found mg/tab 2.42 % Labeled Claim 99.98

Table 3: Results of recovery and precision: Ingredient Amount of drug from formulation 2.5 mg Zolmitriptan 5.0 mg 100 4.0 3.98 98.18 0.78 0.51 100 % Amount Amount % Precision Precision (Intra day) 98.56 1.62 (Inter day) 1.27

Addition added (g/ml) 2.5

recovered Recovery (g/ml) 2.49

Table 4: Accuracy assessment by recovery experiments. Tablet studied zolmitriptan in tablet(g/ml) 2 Zomig-2.5 mg 2 2 Pure zolmitriptan added(g/ml) 1 2 3 Total found(g/ml) 2.96 3.98 4.97 Pure zolmitriptan recovered*in percent 98.66 99.20 99.50

*Mean value of three measurements.

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Calibration Curve of Zolmitriptan in ethanol

1 Absorbance 0.8 0.6 0.4 0.2 0 0 1 2 3 4 5 6 Concentration in mcg/ml

Fig 1: Calibration curve of Zolmitriptan

Absorption Spectrum of Zolmitriptan in ethanol at 226.5 nm

1 Absorbance 0.8 0.6 0.4 0.2 0 210

220

230

240

250

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Wavelength in (nm)

Fig 2: Absorption spectra of Zolmitriptan in ethanol at 226.5 nm.

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