According to the World Health Organization (WHO), cardiovascular disease (CVD) is the leading cause of death in the world every year. The design and development of biosensors for the detection of CVD markers could be one of the major contributions of the scientific community to society. In this context, acetic acid functionalized graphene quantum dots (fGQDs) were used as an interface for the electrochemical detection of cardiac Troponin I (cTnI). The interaction of cTnI with fGQDs for the early diagnosis of acute myocardial infarction was investigated using cyclic voltammetry (CV) and amperometry. The carbodiimide conjugation between the N-H group of cTnI and the functionalized COOH group on GQDs enabled the detection of cTnI biomarker. The same sensing mechanism was confirmed using Fourier Transform Infrared Spectrometry (FTIR). The fGQDs modified Au electrode showed remarkable electrocatalytic oxidation of cTnI with good stability and sensitivity over a linear range of 0.17 to 3 ng mL−1 and a low detection limit of 0.02 ng mL−1. Bland-Altman plots substantiate a bias between the intra-/inter-cTnI assay and calibrated cTnI assay with 95% limits of agreement (mean difference ± 1.96 SD). The aim of this study is to describe an innovative method to detect cardiac biomarker cTnI and provide preliminary data on its diagnostic capacity. At the same time, its applicability in clinical setting will have to be validated with a significant number of samples collected from patients.