ANALYITCAL METHOD DEVELOPMENT AND VALIDATION OF A REVERSED-PHASE HIGHPERFORMANCE LIQUID CHROMATOGRAPHY FOR THE DETERMINATION OF MODAFINIL IN BULK AND PHARMACEUTICAL DOSAGE FORMS

Objective: Todevelopment and validation of a reversed-phase high-performance liquid chromatography (RP-HPLC) for the determination of modafinil in bulk and pharmaceutical dosage forms.Methods: A simple, precise, rapid, and accurate RP-HPLC method was developed for the estimation of modafinil in bulk and pharmaceutical dosage forms. Xterra RP 18 (250 mm × 4.6 mm, 5 μ particle size) with a mobile phase consisting of methanol:water 70:30 V/V was used. The flow rate 1.0 ml/min and the effluents were monitored at 260 nm. The retention time and recovery time was 12 minutes. The detector response was linear in the concentration of 10-50 μg/ml. The respective linear regression equation being Y=452.1x+65237. The limit of detection and limit of quantification were 4.547 and 1.377 mcg, respectively. The method was validated by determining its accuracy, precision, and system suitability.Result: The objective of the present work is to develop simple, precise, and reliable HPLC method for the analysis of modafinil in bulk and pharmaceutical dosage forms. This is achieved using the most commonly employed Xterra RP 18 (250 mm × 4.6 mm, 5 μ particle size) column detection at 260 nm. The present method was validated according to ICH guidelines.Conclusion: In this study, a simple, fast and reliable HPLC method was developed and validated for the determination of modafinil in pharmaceutical formulations.Keywords: Modafinil, Reversed-phase high-performance liquid chromatography, Estimation, ICH guidelines, Tablets.INTRODUCTIONModafinil belongs to the class narcoleptics. The chemical name is 2-[(Diphenylmethyl)-sulfinyl] acetamide. This is a1-adrenergic agonist. Clinical evaluation in hypersomnia and narcolepsy. It is not official in any of the pharmacopoeia. It is listed in the Merck Index 13th edition [1] and Martindale the complete drug reference 36th edition [2]. Literature survey revealed estimation of modafinil by several techniques such as simultaneous estimation by high-performance liquid chromatography (HPLC) [3], determination of modafinil in human plasma by solid-phase and liquid-liquid extraction by HPLC [4-6] determination of modafinil in human urine by HPLC [7], determination of related substance in modafinil [8], and determination of modafinil by a chiral chromatography [9] methods have been reported. In this study, an attempt was made to develop rapid and economical spectrophotometric and reversed-phase HPLC (RP-HPLC) method for estimation of modafinil in bulk and pharmaceutical formulation with better sensitivity, precision, and accuracy using Xterra RP 18 column and UV-detector.Structure [10]ExperimentalInstrumentationQuantitative HPLC was performed on liquid chromatography, Waters separation 2996, photodiode array detector (PDA) detector module equipped with automatic injector with injection volume 20 μl, and 2693 pump. Xterra RP 18 (250 × 4.6 mm, 5 μ particle size was used. The HPLC system was equipped with Empower Software.Chemicals and solventsModafinil was provided as gift sample by Orchid Pharma Ltd., Alathur, India. The solvents which are used methanol and water of HPLC grade were purchased from Merck Specialities Pvt. Ltd., Mumbai, India. Commercial tablets of modafinil were purchased from local market. HPLC grade water obtained from Milli-Q water purification system was used throughout the study [7].Selection of mobile phase• Trail 1 - The method is carried out with the mobile phase acetonitrile and water in the ratio of 70:30 at the 260 nm. In this method, broad peak is found• Trail 2 - The method is carried out by the mobile phase acetonitrile and water in a ratio of 50:50 at 260 nm. In this method, sharp and narrow peak is not came, so we change the mobile phase• Trail 3 - The method was then carried out with the mobile phase water and methanol in the ratio of about 90:10 at 260 nm. This method does not obey the HPLC parameters• Trail 4 - The method was then carried out with the mobile phase methanol and water in the ratio of about 70:30 at 260 nm. This method gives the sharp and narrow peak and it’s obey the HPLC parameters, so we select this mobile phase for HPLC.Preparation of the mobile phase and diluentsThe mobile phase is prepared by methanol and water at the ratio of 70:30 (v/v). The resultant solutions were thoroughly mixed and filtered through a poly-tetra-fluoro ethanol filter of 0.45 μm pore size using Research Article© 2016 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2016.v9i5.13178