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The combination of Ivabradine (IVA) and Metoprolol (MET) was approved by US-FDA for symptomatic treatment of chronic stable angina pectoris. Hence, a potential analytical method that can simultaneously quantify these two drugs is required. In view of this, a novel and fully validated LC-ESI-MS/MS method has been established for the quantification of IVA and MET in rat plasma. Analytes and their deuterated analogues were quantitatively extracted from rat plasma by protein precipitation technique. The analytes were separated using acetonitrile–water consisting 0.1% orthophosphoric acid buffer (30:70 v/v) as a mobile phase with a flow-rate of 1.0 mL/min and 5 min run time on Waters, X-Bridge-C18 (150× 4.6 mm, 3.5 μm) analytical column. The multiple reaction monitoring transitions, m/z 638.14 → 124.22 for IVA, 498.33 → 110.59 for MET; 644.37 → 130.41 for IVA-D6 and 504.46 → 116.28 for MET-D6 were …