RP-HPLC AND CHEMOMETRIC METHODS FOR THE DETERMINATION OF TWO ANTI-DIABETIC MIXTURES; METFORMIN HYDROCHLORIDE-CANAGLIFLOZIN AND METFORMIN HYDROCHLORIDE-GLICLAZIDE IN THEIR PHARMACEUTICAL FORMULATION
Keywords:Binary mixtures, Metformin-Canagliflozin, Metformin-Gliclazide, Spectrophotometric chemometric assisted methods, RP-HPLC
Objective: To develop and validate novel more sensitive analytical methods for the concurrent quantification of metformin-canagliflozin and metformin-gliclazide in their bulk forms and in their pharmaceutical preparations.
Methods: Two methods were developed based on several chemometric assisted spectrophotometric methods and a Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC). The first method applies different spectrophotometric chemometric assisted methods, including ratio difference, derivative ratio and extended ratio subtraction method, while the second method describes a RP-HPLC separation of metformin hydrochloride-canagliflozin and metformin hydrochloride-gliclazide binary mixtures using a C18 column with a mobile phase consisting of acetonitrile: potassium dihydrogen phosphate (adjusted to pH 3) with sodium lauryl sulphate as additive in the ratio of 30:70 (%v/v) in isocratic elution mode at 1 ml/min.
Results: The proposed methods were able to quantify each of the studied drugs in their binary mixtures with high percentage recoveries in both methods. The spectrophotometric methods were able to quantify each of metformin, canagliflozin and gliclazide in the ranges of 2.0-20.0 μg/ml, 1.5-40.0 μg/ml and 2.0-30.0 μg/ml, respectively. The RP-HPLC method produced well-resolved peaks at a retention time of 3.92, 6.92 and 9.10 min in the concentration ranges of 50.0-300.0 μg/ml, 5.0-50.0 μg/ml and 10.0-100.0 μg/ml for metformin, canagliflozin and gliclazide, respectively. The proposed methods were optimized and validated in accordance to the International Conference of Harmonisation (ICH) guidelines in terms of linearity, LOD, LOQ, precision and accuracy.
Conclusion: The developed methods were found to be sensitive and reproducible methods for the simultaneous determination of anti-diabetic binary mixtures; metformin hydrochloride-canagliflozin and metformin hydrochloride-gliclazide. And thus were successfully employed for the quality control analysis of the pharmaceutical formulations of the studied binary mixtures.
World Health Organization. Global Report on Diabetes. World Helath Organisation Global report on diabetes; 2016.
British Pharmacopoeia Commission. British pharmacopoeia, 1932. Analyst 1933;58:697.
USP US. Pharmacopoeia-national formulary [USP 38 NF 33]. Rockville, Md United States Pharmacopeial Conv; 2015.
Panigrahy U, Reddy A. A novel validated RP-HPLC-DAD method for the simultaneous estimation of metformin hydrochloride and canagliflozin in bulk and pharmaceutical tablet dosage form with forced degradation studies. Orient J Chem 2015;3:1489–507.
Reddy NP, Chevela NT. RP-HPLC method development and validation for the simultaneous estimation of metformin and canagliflozin in tablet dosage form. Int J Pharma Sci 2015;5:1155–9.
Gaware D, Patil RN, Harole M. A validated stability indicating RP-HPLC method for simultaneous determination of metformin and canagliflozin in pharmaceutical formulation. World J Pharm Pharm Sci 2015;4:631–40.
Sai VM, Guptha DV, Krishna BSM, Vasantharaju SG. Stability indicating assay method development and validation of naratriptan hydrochloride by RP-HPLC. Res J Pharm Technol 2016;9:1177.
Lotfy HM, Mohamed D, Elshahed MS. Different mathematical processing of absorption, ratio and derivative spectra for quantification of mixtures containing minor component: an application to the analysis of the recently co-formulated antidiabetic drugs; canagliflozin and metformin. Spectrochim Acta Part A Mol Biomol Spectrosc 2018;189:100–9.
Havele SS, Dhaneshwar SR. Simultaneous determination of metformin hydrochloride in its multicomponent dosage forms with sulfonyl ureas like gliclazide and glimepiride using HPTLC. J Liq Chromatogr Relat Technol 2011;34:966–80.
Patil V, Kale S, Sahare P, Vithaldas S. Simultaneous HPTLC analysis of gliclazide and metformin hydrochloride in bulk and tablet dosage form. J Sci Innov Res 2014;3:325–31.
Zhong G, Bi H, Zhou S, Chen X, Huang M. Simultaneous determination of metformin and gliclazide in human plasma by liquid chromatography-tandem mass spectrometry: application to a bioequivalence study of two formulations in healthy volunteers. J Mass Spectrom 2005;40:1462–71.
Agrawal Y, Gogoi P, Manna K, Bhatt H, Jain V. A supercritical fluid chromatography/tandem mass spectrometry method for the simultaneous quantification of metformin and gliclazide in human plasma. Indian J Pharm Sci. 2010;72:50.
Loni AB, Ghante MR, Sawant SD. Simultaneous UV spectrophotometric method for estimation of sitagliptin phosphate and metformin hydrochloride in bulk and tablet dosage form. Der Pharma Chem 2012;4:854-9.
Bhaskar R, Bhaskar R, K Sagar M. UV-spectrophotometric-assisted chemometric methods for the simultaneous determination of metformin hydrochloride and gliclazide in pharmaceutical formulations. Pharm Anal Acta 2012;3. Doi:10.4172/2153-2435.1000158
Bhaskar R, Bhaskar R, Sagar MK, Saini V. Multivariate chemometric assisted analysis of metformin hydrochloride, gliclazide and pioglitazone hydrochloride in bulk drug and dosage forms. Adv Pharm Bull 2013;3:79–84.
Chopade JR, Deshpande SV, Shah S. Simultaneous estimation of metformin HCl and gliclazide by q-analysis method. Int J Pharm Res Sch 2013;2:66–73.
Moosasalam B, Kanaka Narasimhaiah T KVL. A new validated uv spectrophotometric method for the simultaneous estimation of metformin hydrochloride and gliclazide in combined tablet dosage form by dual wavelength method. Inven Rapid Pharm Anal Qual Assur; 2014. p. 1–6.
Havele S, Dhaneshwar S. Development and validation of a hplc method for the determination of metformin hydrochloride, gliclazide and piogliglitazone hydrochloride in multicomponent formulation. Webmed Central Pharm Sci; 2010. p. 1.
Rao U. Determination of gliclazide in a tablet dosage form in the presence of metformin hydrochloride by ion pair reversed phase liquid chromatographic technique. African J Pharm Pharmacol 2011;5:1331–7.
Vadnere BV. RP-HPLC method for simultaneous estimation of gliclazide and metformin in their dosage form. J Pharm Res 2012;5:5036–8.
Sasi E, Goud K, Reddy VK, Sekhar CK. A new simple RP-HPLC method for simultaneous estimation of metformin HCL and gliclazide tablet dosage form. Int J Pharm Biol Sci 2012;2:277–83.
Akula A, Prajwala N, Sandhya M, Maheswara Rao U. Development and validation of RP-HPLC method for simultaneous estimation of metformin hydrochloride and gliclazide in bulk and combined doasage form. Int J Pharm Pharm Sci 2013;5:511–7.
Elkady E, El-Zaher A, Elwy H Mohamed, Saleh M. Validated liquid chromatographic method for simultaneous determination of metformin, pioglitazone, sitagliptin, repaglinide, glibenclamide and gliclazide-application for counterfeit drug analysis. J Anal Bioanal Tech 2015;6. Doi:10.4172/2155-9872.
Nirupama D, Rao PV, Thangabalan B, Manohar Babu S. Method development and validation for metformin hydrochloride and gliclazide in bulk and pharmaceutical formulation by rp-hplc method. Int J Innov Pharm Sci Res 2014;2:1428–37.
Pawar J, Sonawane S, Chhajed S, Kshirsagar S. Development and validation of RP-HPLC method for simultaneous estimation of metformin HCl and gliclazide. Asian J Pharm Anal 2016;6:151–4.
Nirupama D, Venkateswara Rao P, Thangabalan B MBP. Development and validation of RP-HPLC method for simultaneous estimation of gliclazide and metformin in pure and tablet dosage form. Int J Pharm Anal Res 2014;3:326–33.
Amira H Kamal, SFE-M and SFH. A review on UV spectrophotometric methods for simultaneous multicomponent analysis. Eur J Pharm Med Res 2016;3:348–60.
El Jamal MK, Gazy AA. Analysis of three cardiovascular drugs in their ternary mixture using green analytical methodology of smart spectrophotometric methods and RP-HPLC method. Int J Pharm Pharm Sci 2016;8:243–50.
Belal TS, Daabees HG, Abdel-Khalek MM, Mahrous MS, Khamis MM. New simple spectrophotometric method for determination of the binary mixtures (atorvastatin calcium and ezetimibe; candesartan cilexetil and hydrochlorothiazide) in tablets. J Pharm Anal 2013;3:118–26.
Salinas F. A new spectrophotometric method for quantitative multicomponent analysis resolution of mixtures of salicylic and salicyluric acids. Talanta 1990;37:347–51.
Bhatt NM, Chavada VD, Sanyal M, Shrivastav PS. Manipulating ratio spectra for the spectrophotometric analysis of diclofenac sodium and pantoprazole sodium in laboratory mixtures and tablet formulation. Sci World J 2014:1–10. http://dx.doi.org/ 10.1155/2014/495739.
Lotfy HM, Abdel Monem Hagazy M. Comparative study of novel spectrophotometric methods manipulating ratio spectra: an application on pharmaceutical ternary mixture of omeprazole, tinidazole and clarithromycin. Spectrochim Acta Part A Mol Biomol Spectrosc 2012;96:259–70.
El-Bardicy MG, Lotfy HM, El-Sayed MA, El-Tarras MF. Smart stability-indicating spectrophotometric methods for determination of binary mixtures without prior separation. J AOAC Int 2008;91:299–310.
El-Malla S. Simultaneous determination of tapentadol HCl and paracetamol by ratio-spectra derivative spectrophotometry. World J Pharm Sci 2015;3:1290–7.
ICH. Validation of analytical procedures: text and methodology Q2(R1). International Conference on Harmonization of Techinical Requirements for Registration of Pharmaceuticals for Human Use; 2005.
Analytical Methods Technical Commitee. Validation of chromatographic methods. Cent drug Eval Res Food Drug Adm; 1994.