Stability indicating UFLC-PDA assay for simultaneous determination of antazoline hydrochloride and naphazoline hydrochloride in ophthalmic formulations

Abstract

In the present study, simultaneous determination of antazoline hydrochloride (ANZ) and naphazoline hydrochloride (NFZ) in ophthalmic formulations by newly developed ultra-fast liquid chromatographic (UFLC) method was optimized. UFLC separation was performed with ACE Excel 2 C18-PFP column (2 μm, 2.1 x 100 mm) with 0.6 mL/min flow rate of mobile phase which consisted of acetonitrile/phosphate buffer in the ratio of 60:40 (v/v) at 40 ˚C and pH adjusted to 3.0 with 0.5 % v/v triethylamine. The detection of ANZ and NFZ was performed at a wavelength of 285 nm and injection of samples was 1.0 µL. The runtime for analysis was 4.5 min and retention times of NFZ and ANZ were found to be 0.92 and 1.86 min, respectively. The calibration graph was linear for ANZ and NFZ with r2 greater than 0.998 while repeatability and reproducibility (expressed as RSD %) were lower than 1.28 % and 2.14 % respectively. Comparison of UFLC was done with HPLC, UFLC method had remarked advantages over HPLC as run time reduced by 3.4 fold and solvent consumption decreased 5 times. Moreover, detection limits determined by UFLC were 1.5 fold less than HPLC. Force degradation indicated the complete separation of analytes in the presence of degradation products provided the justification of method specificity. The proposed UFLC method was demonstrated to be simple and rapid for the determination of ANZ and NFZ in ophthalmic solutions providing recoveries in the range between 99.6 and 100.4 %.

Keywords

• Histamine H1
• allergic conjunctivitis
• vasoconstrictor
• UFLC