RP-HPLC Method Development for Simultaneous Estimation of Brimonidine Tartrate and Timolol Maleate in Ophthalmic Preparation
DOI:
https://doi.org/10.37285/ijpsn.2023.16.2.4Abstract
Introduction: A simple, precise, rapid, and reproducible RP-HPLC method is developed for the simultaneous estimation of brimonidine tartrate and timolol maleate present in ophthalmic dosage forms.
Method: Gradient RP-HPLC is carried out at 35℃ on column C18 (25 cm X 4.6 mm, 5 µm) with mobile phase a ACN and buffer (25 ml ACN & 475 ml buffer) and mobile phase B water and ACN (37.5 ml water & 212.5 ml ACN) at a flow rate of 1.2ml/min. detection is carried out using DAD detector at 248 nm. Parameter such as linearity, accuracy, precision, specificity, LOD, LOQ and robustness are studied as per ICH guidelines Q2 (R1).
Results: The linearity range of Brimonidine tartrate and Timolol maleate are 10-50 µg/ml and 4-20 µg/ml. The accuracy for both Brimonidine Tartrate and Timolol Maleate are in range of 100-101%. The RSD for Brimonidine Tartrate and Timolol Maleate are less than 2 for precision, specificity, robustness respectively.
Conclusion: This RP-HPLC method can be highly suitable for analysis of brimonidine tartrate and timolol maleate without interference in combined dosage form.
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Keywords:
Brimonidine tartrate, Timolol maleate, RP-HPLC, Ophthalmic preparation, ICH Q2 (R1), Simultaneous estimation
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Acheampong A, Tang L, Diane D. Measurement of brimonidine concentrations in human plasma by a highly sensitive gas chromatography/ mass spectrometric assay. J Pharm Biomed Anal. 1995; 13: 995- 1002.
Ali M, Khatri A, Munir M, Ghori M. A Stability-Indicating Assay of Brimonidine Tartrate Ophthalmic Solution and Stress Testing Using HILIC. Chromatographia. 2007; 70: 539–544.
Anand M, Fonseca A, Santosh G, Padmanabh D. Development and Validation of High Performance Thin Layer Chromatographic Method for Estimation of Brimonidine Tartrate as Bulk Drug and in Ophthalmic Solutions. Int. J. ChemTech Res. 2010; 2: 1376-1379.
Bhagav P, Deshpande P, Pandey S, Chandran S. Development and Validation of Stability Indicating UV Spectrophotometric Method for the Estimation of Brimonidine Tartrate in Pure Form, Formulations and Preformulation Studies. Der Pharm Let. 2010; 2: 106-122.
Damle M, Zirange A. Derivative spectrophotometric method for estimation of brimonidine tartrate & timolol maleate in combination. World Journal of Pharmaceutical Research. 2018 april24; 7(12): 725-731.
Elshanawane A, Abdelaziz L, Mohram M, Hafez H. Development and Validation of HPLC Method for Simultaneous Estimation of Brimonidine Tartrate and Timolol Maleate in Bulk and Pharmaceutical Dosage Form. J Chromatograph Separate Technique. 2014 Jan; 5: 230.
Erk N. Rapid and sensitive HPLC method for the simultaneous determination of dorzolamide hydrochloride and timolol maleate in eye drops with diode-array and UV detection. Pharmazie. 2003; 58: 491-493.
Erk N. Simultaneous determination of dorzolamide HCL and Timolol maleate in eye drops by two different spectroscopic methods. J Pharm Biomed Anal. 2002; 28: 391-397.
Jain P, Khatal R, Jivani H, Surana S. Development and Validation of TLC-densitometry Method for Simultaneous Estimation of Brimonidine tartrate and Timolol maleate in Bulk and Pharmaceutical Dosage Form. J Chromatograph Separat Techniq. 2011; 2: 113-117.
Jiang S, Chappa S, Proksch J. A Rapid and Sensitive LC/MS/MS Assay for the Quantitation of Brimonidine in Ocular Fluids and Tissues. J Chromatogr B Analyt Technol Biomed Life Sci. 2009; 877: 107-114.
Karamanos N, Lamari F, Katsimpris J, Gartaganis S. Development of an HPLC method for determining the alpha2-adrenergic receptor agonist brimonidine in blood serum and aqueous humor of the eye. Biomed Chromatogr. 1999; 13: 86-88.
Kulkarni S, Amin P. Stability indicating HPTLC determination of timolol maleate as bulk drug and in pharmaceutical preparations. J Pharm Biomed Anal. 2000; 23: 983–987.
Marini R et al. Inter laboratory study of a NACE method for the determination of R-timolol content in S-timolol maleate: assessment of uncertainty. Electrophoresis. 2006; 27: 2386-2399.
Elshanawane A, Abdelaziz L, Mohram M, Hafez H. Development and Validation of HPLC Method for Simultaneous Estimation of Brimonidine Tartrate and Timolol Maleate in Bulk and Pharmaceutical Dosage Form. J Chromatograph Separat Techniq. 2014; 5: 230.
Narendra A, Deepika D, Annapurna MM. Liquid chromatographic method for the analysis of brimonidine in ophthalmic formulations. E-Journal of Chemistry. 2012; 9(3): 1327-1331.
Nasir F, Iqbal Z, Khan A, Ahmad L, Shah Y, et al. Simultaneous determination of timolol maleate, rosuvastatin calcium and diclofenac sodium in pharmaceuticals and physiological fluids using HPLC-UV. J Chromatogr B Analyt Technol Biomed Life Sci. 2011; 879: 3434-3443.
Patel A, Asnani A, Vyas A, Patel N, Patel A et al. A brief review on genotoxic impurities in pharmaceuticals. Asian Journal of Pharmaceutical Research. 2021; 11(3):187-193.
Patel A, Prajapati K, Jolapara S, Vyas A, Patel A, Patel N, Pandey M. RP-HPLC method for determination of gemfibrozil using central composite design (CCD). Research Journal of Pharmacy and Technology. 2021; 14(6): 3009-3014.
Patel A, Prajapati K, Vyas A, Patel A, Patel N et al. Determination and validation of phthalate impurities in milk by UV-spectrophotometry method. Pharma Science Monitor 2021; 10(4): 49-58.
Patel A, Ram k, Guttikar S, Vyas A, Patel A, Patel N, Trivedi V. Rapid, sensitive and simple LC-MS/MS method development and validation for estimation of phenytoin in human plasma by using deuterated internal standard. Research Journal of Pharmacy and Technology. 2021; 14(6): 2937-2944.
Patel A; Bundheliya A; Vyas A; Patel N; Patel A et al. A Review on Metal Impurities in Pharmaceutical. Asian Journal of Pharmaceutical Analysis 2021; 11(3): 212-222.
Sharma N, Rao S, Reddy A. A Novel and Rapid Validated Stability Indicating UPLC Method of Related Substances for Dorzolamide Hydrochloride and Timolol Maleate in Ophthalmic Dosage Form. J Chromatogr Sci.2012; 50: 745- 755.
Shirke R, Pai N RP-HPLC determination of brimonidinetartrate in brimonidine tartrate eye drops. Ind Drugs. 2002; 39: 484-486.
Sonanis M, Rajput A. Development and validation of a new stability indicating analytical method for the determination of related components of brimonidine tartrate in drug substances and drug product using UPLC. Int. J. Pharm Pharm Sci. 2011; 3: 156-160.
Turkdemir M, Erdogdu G, Aydemir T, Karagozler A. Voltametric Determination of Timolol Maleate: A β-Adrenergic Blocking Agent. J Anal. Chem. 2001; 56: 1047-1050.
Tzovolou D, Lamari F, Mela E, Gartaganis S, Karamanos N. Capillary electrophoretic analysis of brimonidine in aqueous humor of the eye and blood sera and relation of its levels with intraocular pressure. Biomed Chromatogr. 2000; 14: 301-305.
Vaghela A, Patel A, Patel A, Vyas A, Patel N. Sample Preparation In Bioanalysis: A Review. International Journal of Scientific & Technology Research. 2016; 5(5): 6-10.
Vyas A, Godhaniya J, Patel A, Patel A, Patel N, Chudasama A, Shah S; et al. A review on carcinogenic impurities found in marketed drugs and strategies for its determination by analytical methods. Asian Journal of Pharmaceutical Analysis 2021; 11(2), 159-169.
Vyas A, Gol D, Patel A, Patel A, Patel N, Chavda J, Lumbhani A, Chudasama A. Implementing Analytical Quality by Design (AQbD) Approach for Simultaneous Estimation of Tadalafil and Macitentan by RP-HPLC Method. Analytical Chemistry Letters. 2021; 11(4): 539-552.
Vyas A, Jha S, Patel A, Patel A, Shah S et al. Review on simultaneous equation method (Vierodt's method). Asian Journal of Pharmaceutical Analysis. 2022; 12(2): 149-156.
Vyas A, Patel S, Patel A, Patel A, Patel N et al. Stability testing: An Essential study for Vaccine Formulation Development. Asian Journal of Pharmaceutical Analysis. 2022; 12(1): 29-6.
Vyas A, Rajput M, Patel N, Chotaliya U, Patel A, Patel A; Determination of genotoxic Impurity By Chromatographic Method. An International Journal of Pharmaceutical Sciences 2017; 8(2): 24-31.
Vyas A, Visana N, Patel A, Patel A, Patel N et al. Analytical Quality by Design in Stress Testing or Stability - Indicating Method. Asian Journal of Pharmaceutical Analysis. 2021; 11(2): 170-178.
