A novel, precise, accurate, rapid and cost effective isocratic reverse phase high performance liquid chromatographic (RP-HPLC) method was developed, optimized and validated for the estimation of Netupitant (NET) and Palonosetron (PAL) in bulk and pharmaceutical dosage forms. The drugs were estimated using Phenomenex Gemini C18 (4.6mm×150mm, 5µm) particle size column. A mobile phase composed of tri ethylamine buffer and methanol in proportion of 32:68 v/v, at a flow rate of 1.0 ml/min was used for the separation. Detection was carried out at 248nm. The linearity range obtained was 30-70µg/ml for Netupitant and 10-50µg/ml for Palonosetron with retention times (Rt) of 3.297min and 5.405min for Netupitant and Palonosetron respectively. The correlation coefficient values were found to be 0.999 & 0.999. Precession studies showed % RSD values less than 2 % for both the drugs in all the selected concentrations. The percentage recoveries of Netupitant (NET) and Palonosetron (PAL) were found to be 100.1873% for Netupitant and 100.748% for Palonosetron respectively. The assay results of Netupitant (NET) and Palonosetron (PAL) were found to be 99.82%. The limit of detection (LOD) and limit of quantification (LOQ) were 2.6µg/ml and 7.8µg/ml for Netupitant and 3.4µg/ml 10.2µg/ml for Palonosetron respectively. The proposed method was validated as per the International Conference on Harmonization (ICH) guidelines. The proposed validated method was successfully used for the quantitative analysis of commercially available dosage form. Keywords: Netupitant and Palonosetron, RP-HPLC, ICH Guidelines, Validation.