An isocratic chiral phase high-performance liquid chromatographic method has been developed and validated to quantitation the (R)-isomer in Anagliptin. Separation was achieved in a Lux Cellulose-3 (250×4.6mm, 5μm) column. The ratio of hexane, ethanol and diethyl amine in the mobile phase were optimized to obtain the best separation. UV detection was performed at 254 nm. The described method is linear over a range of LOQ – 1.4 μg/mL of (R)-isomer. The mean recovery of (R)-isomer was found to be in the range of 100–102%. The method is simple, accurate, selective and precise. The method can be used in the quality control of bulk manufacturing.