Method development and validation of HPLC tandem/mass spectrometry for quantification of perindopril arginine and amlodipine besylate combination in bulk and pharmaceutical formulations

Kalaiyarasi Duraisamy, KS Jaganathan, Marothu Vamsi krishna


A well-characterized and fully validated ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometric (UHPLC-ESI-MS/MS) method was developed to reliably analyze combination of perindopril arginine and amlodipine besylate in bulk and tablet formulations. The chromatographic separation was achieved on a Waters ACQUITY UPLC® BEH C18 column with 1.7 μm particle packing which enabled the higher peak capacity, greater resolution, increased sensitivity, and higher speed of analysis using a volatile mobile phase ideally being at least 2 pH units below and above the perindopril arginine and amlodipine besylate pKa, respectively. Mass spectrometric detection was performed using electrospray ion source in positive ion polarity to profile the abundances of perindopril arginine and amlodipine besylate, using the transitions m/z 369 → m/z 172, and m/z 409 → m/z 238 for perindopril arginine and amlodipine besylate, respectively. Calibration curve was constructed over the range 0.25 – 500 ng/mL and 1.0 – 100 ng/mL for perindopril arginine and amlodipine besylate, respectively. The method was precise and accurate, and provided recovery rates > 80% for both compounds. Furthermore, the intra- and inter-assay precision in terms of % RSD was in between 0.1 – 3.7 for both perindopril arginine and amlodipine besylate. A specific, accurate, and precise UHPLC-MS/MS method for the determination of perindopril arginine and amlodipine besylate in bulk and tablet formulation.


Tandem mass spectrometry; Perindopril; Amlodipine

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Gumieniczek A, Mączka P, Inglot T, Pietraś R, Lewkut E, Perczak K. New HPLC method for in vitro dissolution study of antihypertensive mixture amlodipine and perindopril using an experimental design. Cent Eur J Chem. 2013;11(5):717-724.

Ali NW, Abdelwahab NS, Zaki MM, Abdelkawy M. Validated Chromatographic Methods for simultaneous determination of amlodipine besylate and perindopril arginine in binary mixtures and in pharmaceutical dosage form. J Chromatogr Sep Tech. 2012;3(4):134-139.

Gumustas M, Ozkan SA. A validated stability-indicating RP-LC method for the simultaneous determination of amlodipine and perindopril in tablet dosage form and their stress degradation behavior under ICH-recommended stress conditions. J AOAC Int. 2013;96(4):751-757.

Pradhan KK, Mishra US, Pattnaik S, Mishra D, Panigrahi G, Sahu KC. Method development, validation and stability study of perindopril in bulk and pharmaceutical dosage form by UV-Spectrophotometric method. Int J Pharm Biol Arch. 2011;2(4):1114-1122.

Szabó ZI, Réti ZZ, Gagyi L, Kis EL, Sipos E. Simultaneous quantification of related substances of perindopril tert-butylamine using a novel stability indicating liquid chromatographic method. J Chromatogr Sci. 2015;53(3):424-430.

Chatragadda Nagalakshmi, Avula Prameela Rani, Sunkara Bhawani, Chandra Bala Sekaran. Determination of Angiotensin-1 Converting Enzyme Inhibitor, Perindopri Erbumine, in Bulk and Tablet Dosage Form with HPLC. ACSJ. 2012;2(4): 161-176.

Georgakakou S, Kazanis M, Panderi I. Hydrophilic interaction liquid chromatography/positive ion electrospray ionization mass spectrometry method for the quantification of perindopril and its main metabolite in human plasma. Anal Bioanal Chem. 2010;397(6):2161-2170.

Jain PS, Badreshiya PR, Chalikwar SS, Todarwal AA, Surana SJ. Validation of a dissolution method with RP-HPLC analysis for perindopril erbumine and indapamide combination tablet. Chem Ind & Chem Eng Q. 2015;18(1):19-25.

Marolia BP, Bodiwala KB, Shah SA, Prajapati PB, Satani BH, Desai SA. Development and validation of HPTLC method for simultaneous estimation of amlodipine besylate, hydrochlorothiazide and telmisartan in their combined tablet dosage form. Pharm methods. 2016;7(1):48-53.

Bhadani S, Sellappan M. Development and validation of RP-HPLC method for simultaneous estimation of perindopril erbumine and indapamide in combined dosage form. Am J PharmTech Res. 2013;3(3):703-711.

Jogia H, Khandelwal U, Gandhi T, Singh S, Modi D. Development and validation of a stability-indicating assay method for simultaneous determination of perindopril and indapamide in combined dosage form by reversed-phase high-performance liquid chromatography.J AOAC Int. 2010;93(1):108-115.

Patel DB, Mehta FA, Bhatt KK. Simultaneous estimation of amlodipine besylate and indapamide in a pharmaceutical formulation by a high performance liquid chromatographic (RP-HPLC) method. Sci Pharm. 2012;80(3):581-590.

Thörngren JO, Östervall F, Garle M. A high-throughput multicomponent screening method for diuretics, masking agents, central nervous system (CNS) stimulants and opiates in human urine by UPLC–MS/MS. J. Mass Spectrom. 2008;43(7): 980-992.

Nirogi RV, Kandikere VN, Shukla M, Mudigonda K, Maurya S, Komarneni P. High-throughput quantification of perindopril in human plasma by liquid chromatography/tandem mass spectrometry: application to a bioequivalence study. Rapid Commun Mass Spectrom. 2006;20(12):1864-1870.

Validation of analytical procedures: text and methodology Q2(R1). ICH harmonised tripartite guideline. 2005. pp 1-13.

Souri E, Mosafer A, Tehrani MB. Fourth-order derivative spectrophotometric method for simultaneous determination of pseudoephedrine and naproxen in pharmaceutical dosage forms. Res Pharm Sci. 2016;11(2):93-99.


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