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Repaglinide is a novel prandial glucose regulator (PGR) for the treatment of type 2 diabetes mellitus. Repaglinide is mainly metabolised in the liver by CYP3A4 and CYP2C8 enzymes. The objective of the study is to investigate the effects of the CYP3A4 and CYP2C8 genotypes on the pharmacokinetics of repaglinide in 121 healthy Malaysian subjects. Initially, a new HPLC method using a simple liquid-liquid extraction for the determination of repaglinide in human serum was developed and later validated. Then, PCR methods were optimized to detect CYP3A4 and CYP2C8 genetic polymorphisms among healthy Malaysian subjects. Each subject received 4 mg of oral repaglinide. Six blood samples per individual were taken (0 min, 30 min, 60 min, 120 min, 180 min and 240 min) for repaglinide's serum concentration determination by using HPLC. NPAG was then used to determine population pharmacokinetic parameter values of repaglinide.