A NEW HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SEPARATION AND SIMULTANEOUS QUANTIFICATION OF EPTIFIBATIDE AND ITS IMPURITIES IN PHARMACEUTICAL INJECTION FORMULATION

K. SRI GIRIJA; BIKSHAL BABU KASIMALA; VENKATESWARA RAO ANNA

doi:10.22159/ijap.2021v13i2.39895

Authors

K. SRI GIRIJA Department of Chemistry, Koneru Lakshmaiah Education Foundation, Green Fields, Vaddeswaram, Guntur, Andhra Pradesh, India
BIKSHAL BABU KASIMALA Department of Chemistry, Koneru Lakshmaiah Education Foundation, Green Fields, Vaddeswaram, Guntur, Andhra Pradesh, India
VENKATESWARA RAO ANNA Department of Chemistry, Koneru Lakshmaiah Education Foundation, Green Fields, Vaddeswaram, Guntur, Andhra Pradesh, India

DOI:

https://doi.org/10.22159/ijap.2021v13i2.39895

Keywords:

Eptifibatide, Impurity analysis, HPLC, Forced degradation study, Formulation analysis

Abstract

Objective: The objective of the present study is to develop a stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method for qualitative and quantitative determination of Eptifibatide and its impurities in bulk and pharmaceutical dosage forms.

Methods: The chromatographic separation was carried on Phenomenex Luna C18 column (250 mm×4.6 mm; 5µ id) as stationary phase, methanol and phosphate buffer at pH 6.4 in the ratio of 65:45 (v/v) as mobile phase at flow rate of 1.0 ml/min, Ultra Violet (UV) detection was carried at the wavelength of 236 nm and the analysis was completed with a run time of 15 min.

Results: In the developed conditions, the retention time of Eptifibatide and its impurities 1 and 2 were found to be 3.35, 4.93 and 8.18 min, respectively. The method was validated for system suitability, range of analysis, precision, specificity, stability and robustness. Spiked recovery at 50%, 100% and 150% was carried for both standard and impurities and the acceptable % recovery of 98-102 was observed for Eptifibatide and both impurities studied and the % Relative standard deviation (RSD) in each spiked level was found to be less than 2. Stability tests were done through the exposure of the analyte solution to five different stress conditions i. e expose to 1N Hydrochloric acid (HCl), 1 N Sodium hydroxide (NaOH), 3% Hydrogen peroxide (H₂O₂), 80 °C temperature to UV radiation. In all the degradation conditions, standard drug Eptifibatide was detected along with both the impurities studied and the degradation products were successfully separated. In the formulation analysis, there is no other chromatographic detection of other impurities and formulation excipients.

Conclusion: The developed method was found to be suitable for the quantification of Eptifibatide and can separate and analyse impurities 1 and 2.

Downloads

Download data is not yet available.

References

Phillips DR, Teng W, Arfsten A, Nannizzi Alaimo L, White MM, Longhurst C. Effect of Ca2+on GP IIb/IIIa interactions with integrilin: enhanced GP IIb/IIIa binding and inhibition of platelet aggregationby reductions in the concentration of ionized calcium in plasma anticoagulated with citrate. Circulation 1997;96:1488-94.

Robert Scarborough M. Development of eptifibatide. Am Heart J 1999;138:1093-104.

Eli ILev MD, David Hasdai MD, Erez Scapa MD, Ana Tobar MD, Abid Assali MD, Judith Lahav, et al. Administration of eptifibatide to acute coronary syndrome patients receiving enoxaparin or unfractionated heparin: effect on platelet function and thrombus formation. J Am Coll Cardiol 2004;43:966-71.

Perumal Thiagarajan, Kenneth Wu K. Mechanisms of antithrombotic drugs. Adv Pharmacol 1999;46:297-324.

Tcheng JE. Glycoprotein IIb/IIIa receptor inhibitors: putting the EPIC, IMPACT II, RESTORE, and EPILOG trials into perspective. Am J Cardiol 1996;78:35-40.

Conor O'Shea J, Gail Hafley E, Sally Greenberg, Vic Hasselblad, Todd Lorenz J, Michael Kitt M, et al. Platelet glycoprotein IIb/IIIa integrin blockade with eptifibatide in coronary stent intervention. JAMA 2001;285:2468-73.

Maryam Bavand Savadkouhi, Hossein Vahidi, Abdul Majid Ayatollahi, Shirin Hooshfar, Farzad Kobarfard. RP-HPLC method development and validation for determination of eptifibatide acetate in bulk drug substance and pharmaceutical dosage forms. Iran J Phar Res 2017;16:490-7.

Eman Saad Elzanfaly, Enas Abd Elhakim Amer, Sara Abd, Elbasset Galal, Hala Elsayed Zaazaa. Validated high-performance liquid chromatographic method for stability study of eptifibatide. J Chromatogr Sci 2019;57:1–11.

Sushant Patil S, Saurabh Ganorkar B, Atul Shirkhedkar A. Pharmaceutical analysis of eptifibatide via simple, rapid, economical UV-spectrophotometric methods. J Pharm Technol Res Manag 2015;3:49–55.

Eman Saad Elzanfaly, Enas Abdel Hakim Amer, Sara Abdel Basset Galal, Hala Elsayed Zaazaa. Spectrofluorimetric determination of eptifibatide in human plasma and dosage form. J Lumin 2019;34:64–9.

Ronghua Wang, Debby Feder, Frank Hsieh. Characterization of eptifibatide during drug formulation stability assays. J Pharm Biomed Anal 2003;33:1181-7.

Marie Josee Rocheleau, Elaine Larouche, Cristina Salamu, Mihaela Curca. Impurity profiling and in-process testing of drugs for injection by fast liquid chromatography. J Pharm Anal 2012;2:372–7.

Bikshal BK, Useni RM, Venkateswara RA, Maheshwara RL. Intended high-performance liquid chromatography procedure for the quantification of norfloxacin and its potential impurities in active pharmaceutical ingredient and tablet dosage forms. Thai J Pharm Sci 2018;42:27-36.

Mallu UR, Anna VR, Kasimala BB. Rapid stability-indicating HPLC method for the analysis of leflunomide and its related impurities in bulk drug and formulations. Turk J Pharm Sci 2019;16:457-65.

Prasad SS, Krishna Mohan GV, Naga Babu A. Development of simple and robust RP-HPLC method for determination of everolimus and its impurities in oral solid dosage form. Asian J Chem 2019;31:1002-8.

Palacharla SK, Krishna Mohan GV. HPLC method for determination of aspirin, rosuvastatin, ezetimibe and clopidogrel in combination drug products. Asian J Chem 2019;31:2275-83.

Palacharla SK, Krishna Mohan GV, Naga Babu A. RP-HPLC estimation of bumetanide and its impurities in oral solid dosage form. Asian J Chem 2019;31:2275-83.

Prasad SS, Mohan GVK, Babu AN. A quality by design approach for the development of simple and robust reversed-phase stability indicating hplc method for estimation of ibrutinib and its impurities. Rasayan J Chem 2019;12:1434-45.

Geetha Swarupa P, Prasad KRS, Suresh Babu K, Shanmukha Kumar JV. Development and validation of stability indicating RP-HPLC method for simultaneous estimation of escitalopram and l-methylfolate in bulk and tablet dosage form. Rasayan J Chem 2019;12:2338-47.

Palacharla SK, Krishna Mohan GV. Stability indicating HPLC method for the quantification of cefixime, ornidazole and moxifloxacin in solid dosage forms. Rasayan J Chem 2018;11:1696-714.

Ch JR, Rao TB, Kumar Goud PS, Satish J, Rajashekhar K. Development and validation of liquid chromatography method using the principles of QbD for antimalarials used in Artemisinin-based combination therapy. J Liq Chromatogr Relat Technol 2018;41:955-63.

Bikshal Babu Kasimala, Venkateswara Rao Anna, Useni Reddy Mallu. Stability-indicating reversed-phase HPLC method for the separation and estimation of related impurities of cilnidipine in pharmaceutical formulations Indian drugs 2018;55:41-9.

Awdhut Pimpale, Rajendra Kakde. Stability-indicating method development and validation for the simultaneous estimation of rosuvastatin calcium and clopidogrel bisulfate in pharmaceutical dosage form by reverse-phase high-performance liquid chromatography. Asian J Pharm Clin Res 2020;13:84-90.

Senthil Rajan D, Muruganathan G, Kumutaavalli Shivkumar, Ganesh Thangavel. Development and validation of HPLC method for simultaneous quantification of vasicine, lycyrrhizin and piperine in poly herbal cough syrup. Int J Curr Pharm Res 2020;12:15-9.

ICH Validation of Analytical Procedures: Text and Methodology Q2(R1); 1994. p. 1-13.