An Integrated Pharmaceutical Analysis Utilizing Dual HPTLC For Simultaneous Estimation Of Brinzolamide And Brimonidine Tartrate In Bulk And Ophthalmic Formulation

Background: Brinzolamide and Brimonidine Tartrate are Anti-glaucoma agents. Both are new active substances which are useful only for topical use in the treatment of glaucoma. Methods: Two novel, simple, accurate and reliable High-Performance Thin-Layer Chromatography (HPTLC) methods for simultaneous analysis of Brinzolamide (BRZ) and Brimonidine Tartrate (BT) in bulk and ophthalmic formulation. The first method was normal phase chromatography performed on Precoated silica gel F254 TLC plate, with Dichloroethane: Methanol: Triethylamine (4.2: 0.3:0.5, v/v/v) as a mobile phase. The second method was reversed phase chromatography on RP-18 Silica gel F254S TLC plates, with Acetonitrile: water (3.8:1.2, v/v) as mobile phase. HPTLC quantitation of BRZ and BT was done at 260 nm. Results: The HPTLC method resulted into a compact and well resolved band for BRZ and BT at retention factor (Rf) of 0.39 ± 0.01 and 0.61 ± 0.01 for NP-HPTLC and 0.63 ± 0.01 and 0.43 ± 0.01 RP-HPTLC, respectively. The quantitation by HPTLC method was performed over the concentration range of 300 - 1800 ng/band for BRZ and 100 - 600 ng/band for BT with regression coefficient: r2> 0.99 for both methods. Conclusion: A new, precise, and accurate HPTLC method was developed and validated as per ICH guidelines.