• M. MANORANJANI Department of Chemistry, P B Siddhartha College of Arts and Science, Vijayawada, A. P.



Cilnidipine, Ramipril, Development, Validation, RP-HPLC


Objective: The current study focused on the development, validation of Ramipril and Cilnidipine using HPLC and characterization of forced degradation products using LC-MS/MS.

Methods: Simple, accurate, and precise method has been developed for the simultaneous estimation of Cilnidipine and Ramipril in tablet type of dosage form using X-bridge phenyl column (150x4.6mm, 3.5m) with mobile phase of buffer and Acetonitrile in the 30:70 v/v ratio was pumped through a column with a 1 ml/min flow rate. The buffer used in this process was 0.1 percent tri ethyl amine in 1 lt of water and adjust its pH-2.5 with 0.1 percent ortho phosphoric acid. At ambient temperature, chromatography was isocratically performed and run time was 8 min. 240 nm was the optimized wave length.

Results: Cilnidipine and Ramipril retention times were 2.79 min, 5.11 min. respectively. By injecting the norm six times, device parameters of suitability have been studied and the results were found to be well under the acceptance criteria. This approach offers strong linearity over a range of 2-30 µg/ml Cilnidipine 1-15 µg/ml Ramipril concentrations. The regression coefficient R2 > 0.999 from the calibrated curve implies that the linearity of the system was within the range.

Conclusion: The system was validated by using HPLC in terms of accuracy, linearity, method precision, accuracy, limit of detection, limit of quantification, robustness, degradation and the degradation products were characterized by using LC-MS/MS.


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Mohanakumar Sheyanth, Telinius Niklas, Kelly Benjamin, Hjortdal Vibeke. Reduced Lymphatic Function Predisposes to Calcium Channel Blocker Edema: A Randomized Placebo-Controlled Clinical Trial. Lymphatic Research and Biology. Mary Ann Liebert Inc 2019; 18: 156–65.

Poulter NR, Prabhakaran D, Caulfield M. Hypertension. Lancet 2015; 386: 801–12.

Katsura M, Shibasaki M, Hayashida S, Torigoe F, Tsujimura A, Ohkuma S. Increase in expression of α1 and α2/δ1 subunits of L-type high voltage-gated calcium channels after sustained ethanol exposure in cerebral cortical neurons. J Pharmacol Sci 2006; 102: 221–30.

Justiniano CF, Evans DC, Cook CH. Comorbidity–polypharmacy score: a novel adjunct in post-emergency department trauma triage. J Surg Res 2013; 181: 16–9.

Starfield B, Lemke KW, Bernhardt T, Foldes SS, Forrest CB, Weiner JP. Comorbidity: Implications for the Importance of Primary Care in 'Case' Management. Ann Fam Med 2003; 1: 8–14.

Batra AS, Balaji S. Fetal arrhythmias: Diagnosis and management. Indian Pacing Electrophysiol J 2019; 19: 104–9.

Heist EK, Ruskin JN. Drug-induced arrhythmia. Circulation 2010; 122: 1426–35.

McMurray JJ, Pfeffer MA. Heart failure. Lancet 2005; 365: 1877–89.

von Lueder TG, Atar D, Krum H. Diuretic use in heart failure and outcomes. Clin Pharmacol Ther 2013; 94: 490–8.

Wanner Christoph, Inzucchi Silvio E, Lachin John M, Fitchett David, et al. Empagliflozin and Progression of Kidney Disease in Type 2 Diabetes. N Engl J Med 2016; 375: 323–34.

Jiang Shimin, Wang Yining, Zhang Zheng, Dai Peilin, Yang Yue, Li Wenge. Accuracy of hematuria for predicting non-diabetic renal disease in patients with diabetes and kidney disease: A systematic review and meta-analysis. Diabetes Res Clin Pract 2018; 143: 288–300.

Kathiresan S, Srivastava D. Genetics of human cardiovascular disease. Cell 2012; 148: 1242–57.

Hooper L, Martin N, Jimoh OF, Kirk C, Foster E, Abdelhamid AS. Reduction in saturated fat intake for cardiovascular disease. Cochrane Database Syst Rev 2020; 5: CD011737.

Lewis D W, Dorbad D. The utility of neuroimaging in the evaluation of children with migraine or chronic daily headache who have normal neurological examinations. Headache 2000; 40: 629–32.

Bronstein AM, Lempert T. Management of the patient with chronic dizziness. Restor Neurol Neurosci 2010; 28: 83–90.

James Ernest, Sirmans Susan M, Herbert L. Muncie. Dizziness: Approach to Evaluation and Management. Am Fam Physician 2017; 95: 154–62.

Gibson PG, Ryan NM. Cough pharmacotherapy: current and future status. Expert Opin Pharmacother 2011; 12: 1745–55.

Tapper EB, Lok AS. Use of Liver Imaging and Biopsy in Clinical Practice. N Engl J Med 2017; 377: 756–68.

Bernstein JA, Cremonesi P, Hoffmann TK, Hollingsworth J. Angioedema in the emergency department: a practical guide to differential diagnosis and management. Int J Emerg Med 2017; 10: 15.

Bas M, Adams V, Suvorava T, Niehues T, Hoffmann TK, Kojda G. Nonallergic angioedema: role of bradykinin. Allergy 2007; 62: 842–56.

Kovesdy CP. Updates in hyperkalemia: Outcomes and therapeutic strategies. Rev Endocr Metab Disord 2016; 18: 41–7.

Evans KJ, Greenberg A. Hyperkalemia: A review. J Intensive Care Med 2005; 20: 272–90.

Thomas MC. Diuretics, ACE inhibitors and NSAIDs—the triple whammy. Med J Aust 2000; 172: 184–5

Kumar R, Singh VP, Baker KM. The intracellular renin–angiotensin system: implications in cardiovascular remodeling. Curr Opin Nephrol Hypertens 2008; 17: 168–73.

Swati K, Abhishek P, Sushank S, Bothiraja C, Atmaram P. High-performance liquid chromatography for the simultaneous estimation of cefoperazone and sulbactam in rat plasma and its importance in therapeutic drug monitoring. Int J Pharm Pharm Sci 2020; 12: 92-97.

Potturi Ramadevi, Kantipudi Rambabu. Bio analytical method development and validation for Ezetimibe and Pitavastain and its applications to pharmacokinetic studies in Rabbit plasma by using LCMS/MS. Int J Res Pharm Sci 2020; 11: 7854-62.

Asha Eluru, Surendra Babu K. Bio analytical method development and validation for Aplidine in rat plasma and their pharmacokinetic studies by LCMS. World Journal of Pharmacy and Pharmaceutical Sciences 2019; 8: 1201-9.

Ramchandran D, Anita Kethipalli, Krishnamurthy Mannam. Bio analytical method development and validation of Daunorubicin and Cytrarabine in rat plasma by LC-MS/MS and its application in pharmacokinetic studies. J Pharm Sci & Res 2020; 12: 381-6.

Vijayakumari M, Balasekhar reddy Ch. Stability indicating validated hplc method for the determination of zanubrutinib in bulk and pharmaceutical dosage form. Asian J Pharm Clin Res 13: 2020; 159-62.

Shalini K, Ilango K. Development, Evaluation and RP-HPLC method for simultaneous estimation of Quercetin, Ellagic acid and Kaempferol in a poly herbal formulation. Int J App Pharm 2021; 13: 183-92.

Senthil Rajan D, Muruganathan G, Shivkumar K, Ganesh T. Development and validation of hplc method for simultaneous quantification of vasicine, glycyrrhizin and piperine in poly herbal cough syrup. Int J Curr Pharm Res 2020; 12: 15-19.

Malak Y, Al-Bathish A. A, Gazy M. K. El-Jamal. Rp-hplc and chemometric methods for the determination of two anti-diabetic mixtures; metformin hydrochloride-canagliflozin and metformin hydrochloride-gliclazide in their pharmaceutical formulation. Int J Pharm Pharm Sci 2020; 12: 83-94.

Sri Girija K, Bikshal Babu K, Venkateswara Rao A. A new high performance liquid chromatography method for the separation and simultaneous quantification of Eptifibatide and its impurities in pharmaceutical injection formulation. Int J App Pharm 2021; 13: 165-72.

V L N Balaji Gupta T, Venkateswara Rao B, Kishore Babu B. RP-HPLC (stability indicating) based assay method for the simultaneous estimation of Doravirine, Tenofovir Disoproxil Fumarate and Lamivudine. Int J App Pharm 2021; 13: 153-9.

Malathi S, Arunadevi N. Development and validation of stability-indicating simultaneous estimation of metformin and alogliptin in tablets by high-performance thin layer chromatography. Int J Pharm Pharm Sci 2020; 12: 68-73.

Bhavani P, Prasada Rao K, Mohan S. Novel validated reversed-phase high-performance liquid chromatography method for determination of glucosamine, diacerein, and methyl sulfonyl methane in micro sample rat plasma and its application to pharmacokinetic and dissolution studies. Asian J Pharm Clin Res 2020; 13:50-63.

International conference on the harmonization. ICH harmonized tripartite guideline. Validation of analytical procedures: Text and methodology Q2(R1); 2005.

Naresh Kumar D. S, Patel D. Stability indicating chromatographic method development and validation for the simultaneous estimation of escitalopram oxalate and flupentixol in its pharmaceutical dosage form by HPLC. WJPR 2017; 6:549-66.

Manoranjani M. A study of method development, validation and forced degradation for simultaneous quantification of Cisplatin and Fluorouracil in bulk and pharmaceutical dosage form by RP-HPLC. J Pharm Sci & Res 2021; 13: 155-61.

Mayanka Singh, Manoj Charde, Rajesh Shukla, Rita M.C. Determination of calcipotriene in calcipotriene cream 0.05% w/w by RP-HPLC method development and validation. Research J. Pharm. and Tech 2011; 4: 1219-23.

Hemanth Kumar A K, Sudha V, Vijayakumar A, Padmapriyadarsini C. Simultaneous method for the estimation of Bidaquiline and Delamanid in human plasma using high performance liquid chromatography. Int J Pharm Pharm Sci 2021; 13: 36-40.

Shanmugasundaram P, Kamarapu S K. RP-HPLC method for the simultaneous estimation and validation of amlodipine besylate and atenolol in bulk and tablet dosage form in biorelevant dissolution medium (Fassif). Research J Pharm and Tech 2017; 10: 3379-85.

Sruthi A, Uttam Prasad P. Stability indicating method development and validation of Fimasartan by reverse phase high performance liquid chromatography in bulk and pharmaceutical dosage form. Asian J Pharm Clin Res 2021; 14: 138-46.

Gadhvi M.P, Bhandari A, Suhagia B.N, Desai U.H. Development and validation of RP-HPLC method for simultaneous estimation of atazanavir and ritonavir in their combined tablet dosage form. Research J. Pharm. and Tech 2013; 6:200-203.

Narasimha S. Lakka, Chandrasekar Kuppan, Kona S. Srinivas, Raviteja Yarra. Separation and characterization of new forced degradation products of Dasatinib in tablet dosage formulation using LC-MS/MS and stability indicating HPLC methods. Chromatographia 2020; 83: 947-62.

Ashutosh kumar S, Manidipa Debnath, Dr. Seshagiri Rao J.V.L.N, Dr. Gowri Sankar D. Development and validation of a sensitive RP-HPLC method for simultaneous estimation of rosuvastatin and fenofibrate in tablet dosage form by using PDA detector in Gradient mode. Research J. Pharm. and Tech 2016; 9: 549-54.

Balasaheb B Chavan, Vijaya jyothi P, Pradipbhai D Kalariya, et al. Alcaftadine: Selective separation and characterization of degradation products by LC-QTOF-MS/MS. Chromatographia 2018; 81: 631-8.



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