A NEW STABILITY INDICATING UPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF DULOXETINE AND MECOBALAMIN IN BULK AND IN ITS DOSAGE FORMS
Keywords:RP-UPLC, ICH, Duloxetine, Mecobalamin, UV-Spectroscopy, LOD, LOQ
Objective: The objective of the work is to develop and validate a stability indicating, new, simple, highly sensitive RP-UPLC method for simultaneous estimation of Duloxetine and Mecobalamin in bulk and in its dosage forms.
Methods: The Method was developed and excellent chromatographic separation was obtained on a reversed-phase Acquity UPLC BEH C18 (1.0 Ã— 100 mm, 1.7 Âµm) column using an isocratic elution mode by the mobile phase using Methanol: Water (55:45). The flow rate of the mobile phase was 1.0mL/min and the total run time was 10 minutes. UV-Spectroscopic detection at a wavelength of 320 nm was performed and the column oven temperature was maintained at 400C. The analytical procedure was validated by assessing the specificity, linearity, precision, robustness, ruggedness, limit of detection, limit of quantification and accuracy as per ICH guidelines.
Results: The retention times in the standard solution having the concentration of 3 Î¼g/ml and 75 Î¼g/ml of Duloxetine & Mecobalamin were found to be around 4.320 and 5.320 min respectively. The percentage purity values are 99.71 % w/v and 99.02% w/v. Calibration plots were linear (r2 > 0.999) over the concentration range of 1.5 â€“ 5.25 Î¼g/ml and 37.5 â€“ 131.25 Î¼g/ml, the percentage recovery was found to be 99.74% and 99.82% for Duloxetine & Mecobalamin respectively. %RSD for system precision, method precision, robustness and ruggedness was found to be with in 2. The LOD 0.04791 Âµg/mL for Duloxetine and 0.4496 Âµg/mL for Mecobalamin. The LOQ was found to be 0.1452 Âµg/mL for Duloxetine and 1.3625 Âµg/mL for Mecobalamin.
Conclusion: The method represents a fast analytical procedure for the simultaneous quantification of Duloxetine & Mecobalamin. The developed method requires lesser analysis time and less mobile phase consumption. No interference from any component of pharmaceutical dosage form was observed. Validation studies revealed that the method is specific, rapid, reliable, accurate, robust and reproducible. The method is amenable to the routine analysis of large numbers of samples with good precision and accuracy. The most striking feature of this method is its simplicity and rapidity, non-requiring-consuming sample preparation such as extraction with solvents, heating, degassing which are needed for HPLC procedure.
Bassett J, Denny RC, Jeffry GH, Mandham J. Vogelâ€™s Text book of quantitative inorganic analysis. 2nd Edition; 1986. p. 234â€“54.
Satinder Ahuja, Henrik Rasmussen. HPLC method development for pharmaceuticals, 5th Edition. Elsevier; 2009. p. 8-14.
Sharma BK. Instrumental methods of chemical analysis. 25th edition, Goel Publishing House; 2006. p. 286-8.
Usmangani K Chhalotiya, Kashyap K Bhatt, Dimal A Shah, Sunil L Baldania. Development and validation of a stability-indicating RP-HPLC method for duloxetine hydrochloride in its bulk and tablet dosage form. Sci Pharm 2010;78:857-68.
Tapas Kumar Laha, Gitanjali Mishra, Subrata Sen. A validated stability indicating reversed phase high performance liquid chromatographic method of duloxetine and characterization of its degradation products through retro-synthesis. Open Access Sci Rep 2012;1:6.
Srinivasulu Dasari, Raja kumar Viriyala, K Santosh, Archana Kumari NSS D, Ravikumar BVV, SPS Bisht. A validated RP-HPLC method for the analysis of Duloxetine Hydrochloride in pharmaceutical dosage forms. Pharm Globale Int J Compr Pharm 2013;1:3.
Dantu Durga Rao, Shakil S Sait1, A Malleswara Reddy, Dinesh Chakole1, Y Ramakoti Reddy, and K Mukkanti. Analysis of duloxetine hydrochloride and its related compounds in pharmaceutical dosage forms and in vitro dissolution studies by stability indicating UPLC. J Chromatogr Sci 2010;48(10):819-24.
Brunton LL, Parker KS, Lazo JS, Goodman, Gillman's. The Pharmacological Basis of Therapeutics, Mcgraw Hill Publishing; Edition; 2005. p. 436-50.
Saravanan J, Shajan A, Joshi NH, Varatharajan R, Valliappan KA. Simple and validated RP-HPLC method for the estimation of methylcobalamin in bulk and capsule dosage form. Int J Chem Pharm Sci 2010;1:2.
Sharma MC, Sharma AD. Simultaneous estimation and validation of gabapentin and methylcobalamin in tablet dosage form, Hydrotropic approach. Drug Invent Today 2011;3(6):95.
Varsha R Galande, Baheti KG, M Dehghan MH. UV-Vis Spectrophotometric method for estimation of gabapentin and methylcobalamin in bulk and tablet. Int J Chem Res 2010;2(1):695-9.
Pratik Tala, Bipinpaghadar, Komal Dhudashia, Trusha Bhaliya. Analytical method development and validation for simultaneous estimation of duloxetine hydrochloride and methylcobalamin in their pharmaceutical dosage form by absorption correction method. IJPRNS.2013;2(2):213-20.
Sengamalam, R Ravindran, M Gunjan, Manish, Meena S. Analytical method development and dissolution profile of duloxetine and methylcobalamin by Vierodt's method. J Pharm Res 2011;4(2):449.
Pratik Pravinbhai Goti, Parula B Patel. development and validation of ratio-derivative spectrophotometric method for simultaneous estimation of gabapentin, methylcobalamin and alpha lipoic acid in tablet formulation. J Pharm Res 2013;6:609-14.