Single quad mass analyzer coupled UPLC method for impurity profile of Brimonidine tartrate and Timolol maleate: Application in their binary mixture ophthalmic formulation

Abstract

The main objective of the study is to develop a suitable and rapid UPLC/PDA method by coupling online to Quadrupole Dalton analyzer (QDa), a mass detector for the identification and impurity profiling of Brimonidine Tartrate (BRIM)/Timolol maleate (TIMO) in the ophthalmic formulation. Chromatographic separation was achieved on ethylene bridged hybrid octadecylsilane column having 1.7μm particle size in gradient mode using high pure heptafluorobutyric acid as a buffer (A) and water, methanol, and acetonitrile (B) as mobile phase with a flow rate of 0.3mlmin^-1. Based on the spectral maxima, BRIM and its impurities were monitored at 248nm, and TIMO and its impurities were monitored at 295nm. During evaluation of stress conditions and stability data unknown degradants are observed and identified as m/z 218.01 (DP1) and m/z 390.03 (DP2) using QDa-ESI+ scanning mode technique. The performance of the method was systematically validated according to ICH Q2 (R1) guidelines and the method shown very good sensitivity (≥0.5μg.mL^-1) and linearity (r²≥0.999) with consistent recoveries and less than 5% RSD for all compounds. Hence, the proposed UPLC/PDA/QDa method is a simple, sensitive and comprehensive technique where identification and quantification can be done. It gives for complete impurity profile evaluation of BRIM/TIMO in the ophthalmic formulation during quality control in the pharmaceutical industry.

Keywords: Brimonidine tartrate/timolol maleate; Identification; Impurity profiling; Ophthalmic solution; Profil des impuretés; Solution ophtalmique; Tartrate de brimonidine / maléate de timolol; UPLC-PDA-QDa; Validation.