Objective: This study aimed to analyze the metformin hydrochloride in dried blood spot (DBS) sample and evaluate a pharmacokinetic profile in sixhealthy subjects who administered 850-mg metformin hydrochloride in tablet using high-performance liquid chromatography–photodiode array(HPLC–PDA).Methods: Metformin analysis was performed on data of six healthy subjects who administered 850 mg of metformin hydrochloride as tablets. Bloodsamples were taken at 12-time points at intervals of up to 12 h, and data were analyzed using HPLC–PDA.Results: A linear calibration curve was obtained in the range of 25–5000 ng/mL, with r=0.9990. Pharmacokinetic profiles of metformin were obtainedusing DBS samples from six healthy participants, with a Cmax of 347.3–416.22 ng/mL. The average tmax and t1/2 were obtained at 3 and 2 h, respectively,and the area under the curve (AUC)0-t/AUC0-∞ ratio was >80% for all subjects.Conclusion: The DBS biosampling technique can be developed for application in pharmacokinetic study of metformin HCl.