NOVEL SIMULTANEOUSSTABILITY-INDICATING ESTIMATION METHOD FOR DORZOLAMIDE HYDROCHLORIDE AND TIMOLOL MALEATE BYRP-HPLC: APPLICATIONTOAPIANDPHARMACEUTICAL DOSAGE FORMS

Authors

  • DIVYA PINGILI Department of Pharmaceutical Chemistry, Sri Venkateshwara College of Pharmacy, Madhapur, Hyderabad, Telangana--500081, India https://orcid.org/0000-0001-5951-8910
  • SHWETA BHUTADA Department of Pharmacognosy, Sri Venkateshwara College of Pharmacy, Madhapur, Hyderabad, Telangana-500081, India https://orcid.org/0000-0002-6093-0811
  • GUMTE VAISHNAVI Department of Pharmaceutical Analysis, Sri Venkateshwara College of Pharmacy, Madhapur, Hyderabad, Telangana--500081, India
  • ARCHANA AWASTHI Department of Pharmaceutical Chemistry, Sri Venkateshwara College of Pharmacy, Madhapur, Hyderabad, Telangana--500081, India
  • SARITA CHAURASIA Department of Pharmaceutical Chemistry, Sri Venkateshwara College of Pharmacy, Madhapur, Hyderabad, Telangana--500081, India

DOI:

https://doi.org/10.22159/ijpps.2026v18i4.57658

Keywords:

Dorzolamide hydrochloride, Timolol maleate, RP-HPLC, Stability-indicating method, ICH Q2(R1)

Abstract

Objective: To develop a novel stability-indicating validated method for the simultaneous determination of dorzolamide hydrochloride (DH) and timolol maleate (TM) in API and pharmaceuticals.

Methods: Reverse-phase chromatographic separation was achieved using a Spursil C18 column with a mixture of methanol and potassium dihydrogen phosphate buffer (7:3v/v) as the mobile phase at pH 3.5. The flow rate was maintained at1.0 ml/min. DH and TM were detected at 290 nm. The method was validated for linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and forced degradation studies.

Results: The developed method achieved effective separation, with DH and TM eluting at retention times of 2.189 and 3.136 min for standards, and 2.299 min and 3.091 min for samples, respectively. The method showed excellent linearity across 20–100 µg/ml for DH (r² = 0.999) and 5–25 µg/ml for TM (r2 = 0.9993). The mean recovery percentages were 99.99% for DH and 98.79% for TM, with a percentage relative standard deviation (%RSD) below 2.0, confirming the accuracy and precision of the method. Low LOD and LOQ values indicated high sensitivity, while forced degradation studies confirmed the method's stability, indicating capability.

Conclusion: The validated procedure is reliable, accurate, sensitive, and precise for the simultaneous qualitative analysis and stability assessment of DH and TM in both bulk drugs and commercial pharmaceutical formulations. This method is suitable for routine standard quality control applications.

Downloads

Download data is not yet available.

References

1. Sugrue MF. Review the preclinical pharmacology of Dorzolamide HCl, a topical carbonic anhydrase inhibitor. Journal of Ocular Pharmacology and Therapeutics. 1996;12(3):363-76.

2. Curran MP, Orman JS. Bimatoprost/timolol: a review of its use in glaucoma and ocular hypertension. Drugs & aging. 2009 Feb;26(2):169-184.

3. Saccà SC, La Maestra S, Micale RT, Larghero P, Travaini G, Baluce B, Izzotti A. Ability of Dorzolamide HCl and Timolol maleate to target mitochondria in glaucoma therapy. Archives of Ophthalmology. 2011;129(1):48-55.

4. Bell NP, Ramos JL, Feldman RM. Safety, tolerability, and efficacy of fixed combination therapy with Dorzolamide HCl 2% and Timolol maleate 0.5% in glaucoma and ocular hypertension. Clinical Ophthalmology. 2010;4:1331-1346.

5. Fechtner RD, Khouri AS, Realini T. Fixed Combination Drugs. Glaucoma Medical Therapy: Principles and Management. 2020:123.

6. Ormrod D, McClellan K. Topical Dorzolamide HCl 2%/Timolol maleate0.5% A Review of its Use in the Treatment of Open-Angle Glaucoma. Drugs & aging. 2000 ;17(6):477-496.

7. Yeh J, Kravitz D, Francis B. Rational use of the fixed combination of dorzolamide–timolol in the management of raised intraocular pressure and glaucoma. Clinical ophthalmology. 2008 ;2(2):389-399.

8. Mandour AA, Nabil N, Zaazaa HE, Abdelkawy M. Review on analytical studies of some pharmaceutical compounds containing heterocyclic rings: brinzolamide, Timolol maleate, flumethasone pivalate, and clioquinol. Future Journal of Pharmaceutical Sciences. 2020 ;6(1):52.

9. Patel K, Kotadiya R. Advancements in the Analytical Methods for Ripasudil Hydrochloride Hydrate and Timolol Maleate: A Recently Approved FDC. Current Pharmaceutical Analysis. 2023;19(6):487-496.

10. Lotfy HM, Saleh SS. Recent development in ultraviolet spectrophotometry through the last decade (2006–2016): a review. Int J Pharm Pharm Sci. 2016;8(10):40-56.

11. Karpińska J. Derivative spectrophotometry—recent applications and directions of developments. Talanta. 2004;64(4):801-822.

12. Reid IO, Osman SM, Bakheet SM. Comprehensive review of analytical techniques for evaluating fixed-dose combinations containing timolol. Results in Chemistry.2025;7:102309.

13. Gilpin RK, Gilpin CS. Pharmaceuticals and related drugs. Analytical chemistry. 2011 ;83(12):4489-4507.

14. Mandour AA, Nabil N, Zaazaa HE, Ibrahim MM, Ibrahim MA. Two stability-indicating chromatographic methods: TLC densitometric versus HPLC method for the simultaneous determination of brinzolamide and Timolol maleate in ophthalmic formulation in the presence of a probable carcinogenic oxidative degradation product of Timolol maleate. Separations. 2023;10(1):37.

15. Eluru A, Babu KS. A new RP-UPLC method for the separation and simultaneous quantification of dorzolamide HCl and timolol maleate. Int. J. App. Pharm... 2024;16(3):244-250.

16. El Deeb S, Abdelsamad K, Parr MK. Whiter and greener RP-HPLC method for simultaneous determination of Dorzolamide HCl, brinzolamide, and Timolol maleate using isopropanol as a sustainable organic solvent in the mobile phase. Separations. 2024 Mar 12;11(3):83.

17. Annapurna M, Annapurna A, Narendra A. Development and validation of RP-HPLC method for simultaneous determination of Dorzolamide HCl and Timolol maleate in pharmaceutical dosage forms. *Int J Pharm Biomed Res*. 2012;3(1):30–6.

18. Thangabalan B, Kahsay G, Eticha T. New RP‐HPLC method Development and Validation for Dorzolamide HCl in Ophthalmic Dosage Form. Journal of Analytical Methods in Chemistry. 2018;2018(1):4596141.

19. Wanare RS, Kabra AO, Deshmukh A, Aher N. Simultaneous estimation of Dorzolamide HCl and Timolol maleate in eye drops by a RP-HPLC method. Pharmacie Globale. 2012;3(3):1-5.

20. Sharma N, Rao SS, Reddy AM. A novel and rapid validated stability-indicating UPLC method of related substances for Dorzolamide HCl and Timolol maleate in ophthalmic dosage form. Journal of chromatographic science. 2012;50(9):745-755.

21. Sher N, Shafi N, Naseem H, Khan MA. A Novel RP-HPLC method for simultaneous estimation of Dorzolamide HCl and Timolol maleatein ophthalmic products. Pak. J. Chem. 2020;10(1-4):1-6.

22. Shah PA, Gevariya NR, Christian JR, Patel KG, Thakkar VT, Gohel MC, Gandhi TR. Science based development of viscous eye drop of Dorzolamide HCl and Timolol maleate using full factorial design. Pharm methods. 2018; 9:69-78.

23. Erk N. Rapid and sensitive HPLC method for the simultaneous determination of Dorzolamide HCl and Timolol maleatein eye drops with diode-array and UV detection. Die Pharmazie-An International Journal ofPharmaceutical Sciences. 2003;58(7):491-493.

24. 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. Journal of Pharmaceutical Sciences and Research. 202;13(3):155-161.

25. Kirubakaran N, Umashankar MS, Bhuvaneshwari S. Analytical methods of Estimation of Oxetacaine and its quality determinations in different Pharmaceutical Formulations by Reverse Phase High Performance Liquid Chromatography (RP-HPLC). Research Journal of Pharmacy and Technology. 2023;16(12):5748-5753.

26. ICH Q2(R1). Validation of analytical procedures: text and methodology. Geneva: International Council for Harmonisation; 2005.

27. Marina J A, Thanikachalam PV. Development and Validation of QBD-Assisted Using Central Composite Design RP-HPLC Method for LobeglitazoneSulfate and Glimipiride in Bulk and Its Combined Dosage Form. Chromatographia. 2025;88(5):381-393.

28. Gawde O, Nayak S, Vaidhun B. Analysis of Nefopam Hydrochloride from bulk and dosage form using validated RP HPLC method. International Journal of Pharmaceutical Sciences and innovation. 2021;1:40-53.

29. Veneela AS, Rajitha GA, Narikimalli AS. A stability indicating RPHPLC method development and validation for the simultaneous estimation of daunorubicin and cytarabine in bulk and pharmaceutical dosage form. Asian J Pharm Clin Res. 2022;15(5):58-63.

30. Katolkar PA, Jaiswal SH. Analytical method development and validation for the estimation of cyamemazine tartrate in formulation by RP-HPLC with stability indicating. Asian J Pharm Clin Res. 2022;15(9):28-32.

31. Ramakrishna K, Kumar S. Development and validation of stability-indicating RP-HPLC method for simultaneous determination of dorzolamide hydrochloride and timolol maleate in ophthalmic dosage form. Int J App Pharm. 2025;17(6):512-525.

32. Deepalatha C, Pratyusha L. To study the drug use pattern in primary open-angle glaucoma in a tertiary care teaching hospital. Asian J Pharm Clin Res. 2017;10(2):198-200.

Published

14-02-2026

How to Cite

PINGILI, DIVYA, et al. “NOVEL SIMULTANEOUSSTABILITY-INDICATING ESTIMATION METHOD FOR DORZOLAMIDE HYDROCHLORIDE AND TIMOLOL MALEATE BYRP-HPLC: APPLICATIONTOAPIANDPHARMACEUTICAL DOSAGE FORMS”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 18, no. 4, Feb. 2026, doi:10.22159/ijpps.2026v18i4.57658.

Issue

Articles In Press [Apr 2026]

Section

Original Article(s)

Copyright (c) 2026 ARCHANA AWASTHI, Divya Pingili, SHWETA BHUTADA, VAISHNAVI GUMTE, SARITA CHAURASIA

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Similar Articles

<< < 5 6 7 8 9 > >> 

You may also start an advanced similarity search for this article.