A new simple isocratic chiral liquid chromatographic method was developed for the enantiomeric purity of 1, 2, 3, 4- tetrahydro-1-naphthoic acid (THNA), a key starting raw material for the synthesis of Palonosetron HCl final drug substance. The chromatographic separation was achieved on Chiralcel OD-H, 250 mm Ð¥ 4.6 mm, 5 μm column using a mobile phase system consisting of n-hexane, isopropyl alcohol and trifluoroacetic acid in the ratio of 948:50:2 (v/v/v). The mobile phase was pumped through column at the flow rate of 0.8 mL min-1. Addition of trifluoroacetic acid in the mobile phase enhanced chromatographic efficiency and resolution between the isomers. The resolution between the isomers was found to be more than three. The developed method was subsequently validated and proved to be an accurate, specific and precise. The experimentally established limit of detection and quantification for (R)- enantiomer were found to be 0.4046μg mL-1 and 0.8186 μg mL-1 respectively and for (S)-THNA were found to be 0.5916μg mL-1 and 0.9860 μg mL-1 respectively for 20 μl injection volumes. The percentage recoveries of (R)- enantiomer were ranged from 98 to 102. The stability of sample in analytical solution was checked for about 36 hours at room temperature and was found to be stable for about 36 hours. The proposed method was found to be suitable and accurate for the quantitative determination of (R)-enantiomer of THNA in (S)-THNA.