Design of Experiment Utilization to Develop a Simple and Robust RP-UPLC Method for Stability Indicating Method of anti glaucoma Ophthalmic Drops

[摘要] Brimonidine Tartrate and Timolol Maleate are used in treatment of glaucoma by decreasing intra ocular pressure. It is necessary to establish a Validated UPLC method for the assay of Brimonidine Tartrate and Timolol Maleate as well as Benzalkonium Chloride in eye drops. Using of DOE techniques provides more information about different factors affects system suitability of new method. DOE led to highly robust methods through creating design space. The method was performed on Phenomenex Kinetex C18 (50x4.6 mm, 2.6 µm) and the mobile phase consisted of Methanol and 1- Decane sulphonic acid sodium (pH 3.0, 0.01M) which pumped at a flow rate 0.6 mLmin-1 at 50 °C in gradient manner. 10 µL of drugs sample solutions were monitored at wavelengths 320 nm.Brimonidine Tartrate, Timolol Maleate and Combigan ED were stressed under different conditions in forced degradation studies. Degradation products had a good resolution with main drugs and Benzalkonium Chloride. The proposed method was validated in terms of linearity, accuracy, precision and limits of detection and quantitation according to ICH.

[学科分类] 分析化学

[关键词] Brimonidine Tartrate; Timolol Maleate; Benzalkonium Chloride;UPLC; Stability Indicating Method; Design of Experiment