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Metformin (MET), an oral biguanide agent, can improve insulin resistance and decrease hepatic glucose production, leading to a reduction in blood-sugar levels. The objective of the present study was to develop and validate simple and rapid LC-MS/MS method for analysis of MET in dried blood spot (DBS) sample for patient monitoring studies purposes (drug adherence). The chromatographic separation was achieved with Waters HSS-T3 column using gradient elution of mobile phases of two solvents 1) solvent A, consisted of 10mM ammonium formate