A simple and specific method for simultaneous determination of Ebastine and Carebastine by liquid chromatography-tandem mass spectrometry operated in positive ionization mode was developed and validated. The column used was a BDS Hypersil C18, 50 mm × 4.6 mm, 5μm with a flow rate of 0.6 mL/min, without splitter for the chromatographic analysis with flow rate of 0.6 mL/min. Solid phase extraction method was applied. Mass parameters, 496.2/261.0 and 672.2/479.3/261.2 were chosen for analysis. Linearity was established in human plasma covering the concentration range of 0.051 ng/mL to 31.099 ng/mL for Ebastine and 1.013 ng/mL to 1005.451 ng/mL for Carebastine using (Ebastine D6 and Carebastine D6) as internal standards. Different parameters such as linearity, range, precision, accuracy, ruggedness and robustness, limit of detection (LOD) and limit of quantification (LOQ) were used for full validation of the method. The results were found to be acceptable as per the guidelines of International Conference on Harmonization (ICH). The method found to be novel, rapid, linear, precise, accurate, robust and rugged and can be successfully applied for the routine analysis of Ebastine and Carebastine with more sensitivity and covers wider range of quantitation. The method also found to be useful and economical.