Abstract
Background: Design of experiment (DOE) is considered the most powerful tool to identify factors that affect variation and improve the response by tuning these factors. In the present work, DOE was applied to establish an innovative, sensitive, and precise HPLC method for the simultaneous determination of Escitalopram oxalate, Paroxetine hydrochloride hemihydrate and Clonazepam in the presence of their related compounds in drug substance and drug products.
Methods: Buffer molarity, % organic modifier (acetonitrile content) at the beginning/end of the gradient, flow rate at the beginning/end of the gradient, pH of the mobile phase, and column temperature were screened using Plackett-Burman design (PBD) model. The main effect plot showed that % organic modifier at the beginning/end of gradient and flow rate at the beginning of gradient were statistically significant variables influencing peaks resolution (p<0.05). Box-Behnken design (BBD) was then used as an optimization model in order to achieve the highest possible resolution with the least possible experimental trials through studying the interaction and quadratic effects of these three factors. Finally, the optimum condition for predicated peak resolution could be achieved by the desirability function.
Results: After optimization, the chromatographic separation was attained on Agilent Zorbax SB C18 (4.6×250mm, 5μm) column using gradient elution of mobile phase: (A) potassium dihydrogen phosphate (pH 2.7; 0.025M) and (B) acetonitrile at ambient column temperature with the last eluted compound in less than 17 min. The flow rate was maintained at 1/2.3 mLmin-1 with UV detection at 245/210 nm using time programming.
Conclusion: The optimized chromatographic method was used for stability, indicating assay of the cited drugs in the presence of their related compounds according to ICH Q2R1 guidelines.
Keywords: Escitalopram oxalate, paroxetine hydrochloride hemihydrates, clonazepam, design of experiment, plackett-Burman design, box-Behnken design.
Graphical Abstract
[1]
Rockville, M.D. The United States Pharmacopeia and National Formulary. The United States Pharmacopeial Convention, Inc., 2019. USP 42–NF 37.
[2]
Ibrahim, F.; El-Enany, N.; Shalan, S.; Elsharawy, R. Micellar high-performance liquid chromatographic method for simultaneous determination of clonazepam and Paroxetine HCl in pharmaceutical preparations using monolithic column. J. Chromatogr. Sep. Tech., 2016, 7, 331.
[http://dx.doi.org/10.4172/2157-7064.1000331]
[http://dx.doi.org/10.4172/2157-7064.1000331]
[3]
Kasper, S.; Stein, D.J.; Loft, H.; Nil, R. Escitalopram in the treatment of social anxiety disorder: randomised, placebo-controlled, flexible-dosage study. Br. J. Psychiatry, 2005, 186, 222-226.
[http://dx.doi.org/10.1192/bjp.186.3.222] [PMID: 15738503]
[http://dx.doi.org/10.1192/bjp.186.3.222] [PMID: 15738503]
[4]
Baldwin, D.S.; Anderson, I.M.; Nutt, D.J.; Bandelow, B.; Bond, A.; Davidson, J.R.; den Boer, J.A.; Fineberg, N.A.; Knapp, M.; Scott, J.; Wittchen, H.U. British Association for Psychopharmacology. Evidence-based guidelines for the pharmacological treatment of anxiety disorders: recommendations from the British Association for Psychopharmacology. J. Psychopharmacol., 2005, 19(6), 567-596.
[http://dx.doi.org/10.1177/0269881105059253] [PMID: 16272179]
[http://dx.doi.org/10.1177/0269881105059253] [PMID: 16272179]
[5]
British Pharmacopoeia Medicines and Health Care Products Regulatory Agency London., 2018, 3, 383-384.
[6]
Yanamadala, G.; Srikumar, P. Development and validation of a stability-indicating HPLC method for the simultaneous determination of Paroxetine hydrochloride and Clonazepam in pharmaceutical dosage forms Int J Pharm., 2014, 4, 448-457. CODEN: IJPNL6.Yanamadala-Srikumar/b72c8fbfd58fc51-ca0371c70019e80e6d2683fc3
[7]
Brown, J.H.; Taylor, P.; Robert, L.J.; Marrow, J.D. Goodman and Gilman’s The Pharmacological Basis of Therapeutics; Mc-Graw-Hill.: New York., 2001, p. 280..
[8]
Dar, S.A.; Bhat, B.A.; Jan, M.M. Addition of benzodiazepines to selective serotonin reuptake inhibitors to optimize treatment of depression: a hospital based study. Int J Res Med Sci., 2018, 6(6), 2081-2085.
[http://dx.doi.org/10.18203/2320-6012.ijrms20182293]
[http://dx.doi.org/10.18203/2320-6012.ijrms20182293]
[9]
Dunlop, B.W.; Davis, P.G.; Paula, G. Combination treatment with benzodiazepines and SSRIs for comorbid anxiety and depression: a review. Prim. Care Companion J. Clin. Psychiatry, 2008, 10(3), 222-228.
[http://dx.doi.org/10.4088/PCC.v10n0307] [PMID: 18615162]
[http://dx.doi.org/10.4088/PCC.v10n0307] [PMID: 18615162]
[10]
Seedat, S.; Stein, M.B. Double-blind, placebo-controlled assessment of combined clonazepam with paroxetine compared with paroxetine monotherapy for generalized social anxiety disorder. J. Clin. Psychiatry, 2004, 65(2), 244-248.
[http://dx.doi.org/10.4088/pcc.v10n0307] [PMID: 15003080]
[http://dx.doi.org/10.4088/pcc.v10n0307] [PMID: 15003080]
[11]
Bhimanadhuni, C.N.; Garikapati, D.R.; Usha, P. Development and validation of an RP-HPLC method for the simultaneous determination of Escitalopram Oxalate and Clonazepam in bulk and its pharmaceutical formulations. Int. Curr. Pharm. J., 2012, 1(8), 193-198.
[http://dx.doi.org/10.3329/icpj.v1i8.11249]
[http://dx.doi.org/10.3329/icpj.v1i8.11249]
[12]
Boss, P.V.S.C.; Anbazhagan, S.; Raja, P.R.; Valarmathy, S.; Winkumar, G.; Suriya, R. RP – HPLC method development and validation for the determination of escitalopram oxalate and clonazepam in tablet dosage form. Int. J. Pharm., 2018, 3(10), 35-44.
[http://dx.doi.org/10.3329/icpJ.V1i8.11249]
[http://dx.doi.org/10.3329/icpJ.V1i8.11249]
[13]
Chakole, R.D.; Charde, M.S.; Bhavsar, N.; Marathe, R.P. Simultaneous estimation of Escitalopram and Clonazepam by RP-HPLC in pharmaceutical formulations Inter. J. Phytopharm., 2012, 2(1), 25-29.
[http://dx.doi.org/10.7439/ijpp.v2i1.437]
[http://dx.doi.org/10.7439/ijpp.v2i1.437]
[14]
Naidu, N.; Yellappa, M.; Praveenkumar, L.; Koteswararao, M.I.P. Assay method development and validation for simultaneous estimation of Paroxetine and Clonazepam bY RP- HPLC. WJPMR., 2018, 4(4), 232-236.www.wjpmr.com
[15]
Sulthana, P.S.; Dhachinamoorhti, D.; Prasada Rao, C.H.M.M. Assay method development and validation for simultaneous estimation of Paroxetine and Clonazepam bY RP- HPLC. EJBPS., 2017, 4, 267-272.https://www.ejbps.com/ejbps/abstract_id/2704
[16]
Unnisa, A.; Kumar, S.; Babu, Y.R.; Chaitanaya, S. Development and validation of RP-HPLC-PDA method for the simultaneous estimation of clonazepam and paroxetine hydrochloride in bulk and tablet dosage forms. J. Pharm. Res., 2014, 8(9), 1212-1217.http://jprsolutions.info/newfiles/journal-file-56c2839e3ee781.92157749.pdf
[17]
Kakde, R.B.; Satone, D.D.; Gadapayale, K.K.; Kakde, M.G. Stability-indicating RP-HPLC method for the simultaneous determination of escitalopram oxalate and clonazepam. J. Chromatogr. Sci., 2012, 7, 1-6.
[http://dx.doi.org/10.1093/chromsci/bms177] [PMID: 23135233]
[http://dx.doi.org/10.1093/chromsci/bms177] [PMID: 23135233]
[18]
Kalia, B.; Baghel, U.S. Method development and validation of stability indicating RP-HPLC method for simultaneous estimation of escitalopram oxalate and clonazepam in bulk and its pharmaceutical formulations. JDDT, 2019, 9(1-s), 265-274.
[http://dx.doi.org/10.22270/jddt.v9i1-s.2347]
[http://dx.doi.org/10.22270/jddt.v9i1-s.2347]
[19]
Reddy, G.S.; Prasad Reddy, S.L.N.; Reddy, L.S.K. Development and Validation of a stability indicating liquid chromatographic method for the simultaneous estimation of Paroxetine and Clonazepam in bulk and its pharmaceutical formulation. Int. J. Pharm. Pharm. Sci., 2014, 6(10), 397-402.https://innovareacademics.in/journals/index.php/ijpps/article/view/1687
[20]
Dhavale, N.; Gandhi, S.; Sabnis, S.; Bothara, K. Simultaneous HPTLC determination of escitalopram oxalate and clonazepamin combined tablets. Chromatographia, 2008, 67, 487-490.
[http://dx.doi.org/10.1365/s10337-008-0524-7]
[http://dx.doi.org/10.1365/s10337-008-0524-7]
[21]
Kakde, R.; Satone, D.; Bawane, N. HPTLC method for simultaneous analysis of Escitalopram and Clonazepam in pharmaceutical preparation. J. Planar Chromatogr. Mod. TLC, 2009, 22, 417-420.
[http://dx.doi.org/10.1556/JPC.22.2009.6.5]
[http://dx.doi.org/10.1556/JPC.22.2009.6.5]
[22]
Shah, P.; Patel, J.; Patel, K.; Gandhi, T. Development and Validation of HPTLC method for simultaneous estimation of Clonazepam and Paroxetine hydrochloride using DOE approach. JTUSCI, 2015, 121-132.
[23]
Gandhi, S.V.; Dhavale, N.D.; Jadhav, V.Y.; Sabnis, S.S. Spectrophotometric and reversed-phase high-performance liquid chromatographic methods for simultaneous determination of escitalopram oxalate and clonazepam in combined tablet dosage form. J AOACInt., 2008, 91, 33-38.francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20081883
[24]
Kakde, R.B.; Satone, D.D. Spectrophotometric method for simultaneous estimation of escitalopram oxalate and clonazepam in tablet dosage form. Indian J. Pharm. Sci., 2009, 71(6), 702-705.
[http://dx.doi.org/10.4103/0250-474X.59559] [PMID: 20376230]
[http://dx.doi.org/10.4103/0250-474X.59559] [PMID: 20376230]
[25]
Patel, J.U.; Dalwadi, H.; Shah, D.R.P. Application of ratio derivative spectrophotometry for simultaneous determination of Clonazepam and Paroxetine hydrochloride in tablet dosage form. JPRR, 2018, 2-8.
[26]
Boda, J. M.; Bhalodiya, H. A.; Patel, P. B. UV spectroscopic method for simultaneous estimation of clonazepam and paroxetine hydrochloride hemihydrate in combined pharmaceutical formulation Inventi Rapid: Pharm Ana QA., 2012. Inventi:PPaqa/333/12
[27]
Patel, J.U. A Study of the novel ratio difference spectrophotometric method for the analysis of Clonazepam and Paroxetine hydrochloride in combined tablet dosage form. Pharm Anal. Quality Assur., 2016, 4, 182-187. Inventi:ppaqa/19534/16
[28]
Sheeja, V.K.; Swapna, A.S.; Eapen, S.C.; Kumar, P. Method development and validation for the simultaneous estimation of clonazepam and paroxetine in combined dosage form using colorimetry. Asian J. Res. Chem, 2014, 7(1), 48-51.
[http://dx.doi.org/10.5958/0974-4150]
[http://dx.doi.org/10.5958/0974-4150]
[29]
Patel, J.U.; Chhalotiya, U.K.; Shah, P.A. A Synchronous fluorescence spectrofluorometric method for the simultaneous determination of Clonazepam and Paroxetine Hydrochloride in combined pharmaceutical dose form. Turk J Pharm Sci, 2017, 14(3), 251-256.
[http://dx.doi.org/10.4274/tjps.84856] [PMID: 32454621]
[http://dx.doi.org/10.4274/tjps.84856] [PMID: 32454621]
[30]
Plackett, R.L.; Burman, J.P. The design of optimum multi factorial experiments. Biometrika, 1946, 33, 305-325.
[http://dx.doi.org/10.1093/biomet/33.4.305]
[http://dx.doi.org/10.1093/biomet/33.4.305]
[32]
Ganorkar, S.B.; Shirkhedkar, A.A. Design of experiments in liquid chromatography (HPLC) analysis of pharmaceuticals: analytics, applications, implications and future prospects. Rev. Anal. Chem., 2017, 36(6)
[http://dx.doi.org/10.1515/revac-2016-0025]
[http://dx.doi.org/10.1515/revac-2016-0025]
[33]
Debrus, B.; Lebrun, P.; Ceccato, A.; Caliaro, G.; Rozet, E.; Nistor, I.; Oprean, R.; Rupérez, F.J.; Barbas, C.; Boulanger, B.; Hubert, P. Application of new methodologies based on design of experiments, independent component analysis and design space for robust optimization in liquid chromatography. Anal. Chim. Acta, 2011, 691(1-2), 33-42.
[http://dx.doi.org/10.1016/j.aca.2011.02.035] [PMID: 21458628]
[http://dx.doi.org/10.1016/j.aca.2011.02.035] [PMID: 21458628]
[34]
Candioti, L.V.; De Zan, M.M.; Cámara, M.S.; Goicoechea, H.C. Experimental design and multiple response optimization. Using the desirability function in analytical methods development. Talanta, 2014, 124, 123-138.
[http://dx.doi.org/10.1016/j.talanta.2014.01.034] [PMID: 24767454]
[http://dx.doi.org/10.1016/j.talanta.2014.01.034] [PMID: 24767454]
[35]
Hibbert, D.B. Experimental design in chromatography: a tutorial review. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2012, 910, 2-13.
[http://dx.doi.org/10.1016/j.jchromb.2012.01.020] [PMID: 22333438]
[http://dx.doi.org/10.1016/j.jchromb.2012.01.020] [PMID: 22333438]
[36]
El-Zaher, A.A.; Mahrouse, M.A. Utility of experimental design in pre-column derivatization for the analysis of tobramycin by HPLC- fluorescence detection application to ophthalmic solution and human plasma. Anal. Chem. Insights, 2013, 8, 9-20.
[http://dx.doi.org/10.4137/ACI.S11612] [PMID: 23700362]
[http://dx.doi.org/10.4137/ACI.S11612] [PMID: 23700362]
[37]
Khamanga, S.M.; Walker, R.B. The use of experimental design in the development of an HPLC-ECD method for the analysis of captopril. Talanta, 2011, 83(3), 1037-1049.
[http://dx.doi.org/10.1016/j.talanta.2010.11.025] [PMID: 21147355]
[http://dx.doi.org/10.1016/j.talanta.2010.11.025] [PMID: 21147355]
[38]
Chaudhari, S.R.; Shirkhedkar, A.A. Desgin of Experiment Avenue for development and validation of RP-HPLC-PDA method for determination of apremilast in bulk and in in-house tablet formulation. J. Anal. Sci. Technol., 2019, 10, 1-10.
[http://dx.doi.org/10.1186/s40543-019-0170-8]
[http://dx.doi.org/10.1186/s40543-019-0170-8]
[39]
Ragonese, R.; Macka, M.; Hughes, J.; Petocz, P. The use of the Box-Behnken experimental design in the optimisation and robustness testing of a capillary electrophoresis method for the analysis of ethambutol hydrochloride in a pharmaceutical formulation. J. Pharm. Biomed. Anal., 2002, 27(6), 995-1007.
[http://dx.doi.org/10.1016/S0731-7085(01)00659-8] [PMID: 11836062]
[http://dx.doi.org/10.1016/S0731-7085(01)00659-8] [PMID: 11836062]
Article Metrics
3
1