A validated rp-HPLC method for simultaneous determination of olopatadine hydrochloride and its related substances in eye drop

Olopatadine Hydrochloride, a H1-receptor antagonist, widely used for Allergic Conjunctivitis. A HPLC method was created and validated for the quantification of olopatadine hydrochloride and its related substances in eye formulation in accordance with ICH standards. The separation of olopatadine hydrochloride was done by using a 4.6-mm 15-cm; 5 mu m packing USP L7 column with a mobile phase made up of a mixer of phosphate buffer, pH 3.0: Acetonitrile (72:28) ratio at a flow rate of 1.0 mL/min and injection volume 30 mu L. A photodiode array detector was used at a wavelength of 299 nm. The validation parameters including specificity, LOD/LOQ, linearity, accuracy, precision, robustness, and solution stability, were verified for performance of the method. The linearity ranges were 0.02-0.40, 0.0004-0.008 and 0.0001-0.002 mg/mL for olopatadine hydrochloride, olopatadine-related compound B and olopatadine carbaldehyde, respectively. The % recovery was within the 80-120%. When the column temperature and flow rate were changed, the system suitability resolution was still within the acceptable range. The solutions were stable after 24 hr at both room temperature and 5 degrees C temperature. For the analysis of olopatadine hydrochloride and its impurity in pharmaceutical finished eye dosage form, the confirmed HPLC method can be a workable analytical approach.