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Introduction and Aim: Mangiferin is accompanied by the therapeutic potential for various human ailments. The present study is aimed to develop a validated HPLC method for the quantification of mangiferin in coffee (Coffea arabica) leaves. Materials and Methods: The ­­high-performance liquid chromatography (HPLC) was carried out on Agilent Technologies 1260 LC Infinity Series liquid chromatography system (Agilent, USA) using an isocratic mobile phase of a mixture of methanol and 1% orthophosphoric acid v/v (70:30) at a flow rate 1mL min-1 for 14 min with the controlled temperature at 25°C. The mangiferin content was detected by a UV diode array detector at 258 nm. Results: The linearity of the HPLC system was established in the range of 10-100 ppm mL-1 with a regression coefficient (R2) of 0.99670. The accuracy of the method was assessed by a study performed using 3 different levels, with a 99% average recovery. The limit of detection(LOD) and limit of quantification (LOQ) were 1.3 and 3.9 ppm mL-1. Intraday and interday precision analysis showed the relative standard deviation 0.012342 separately. The content of mangiferin in the coffee leaves was 94 mg/gram (w/w). Conclusion: All necessary factors, such as linearity, accuracy, LOD, LOQ, precision, and % relative standard deviation(RSD), were inside the appropriate limits. The HPLC was validated using all necessary variables with a successful estimation of mangiferin.