The exposition of minerals to oxygen as well as non-treated tailings in mining activities alter the balance of the ecosystem, specifically leading to the generation of acidic solutions in the presence of sulfidic minerals. Several secondary iron minerals are precipitated in these settings that can be detected via remote sensing applications. The purpose of this research is to investigate the capacity of hyperspectral analysis to determine the abundance of Acid Mine Drainage (AMD)-indicator secondary iron minerals in mine sites with the guidance of ground truth information. To this end, we focus on an abandoned coal mine site in Turkey to detect secondary iron minerals associated with AMD via multispectral Sentinel-2 imagery, in accordance with the laboratory analysis of fieldcollected samples through X-Ray Diffraction (XRD), Inductive Coupled Plasma (ICP), and ASD spectral analysis. In relation with the conducted laboratory XRD and ICP results, the proposed methodology first reveals the iron-induced absorption feature located between 700 and 900 nm on field-collected ASD spectra and reference USGS spectra through a baseline method, namely parabola fitting method. The subsequent remote sensing analysis then applies hyperspectral unmixing to Sentinel-2 imagery and identifies the spectral endmember indicating iron-absorption behavior by computing its spectral angle distance to reference spectra. The experiments reveal that while the iron absorption characteristics are not apparent in pixel spectra, the utilized unmixing methodology enables capturing of those features at sub-pixel level on the resulting endmembers. Second, the comparison between the calculated abundances with unmixing and iron levels obtained with ground based ICP analysis indicate coherent correlation values. Finally, among the utilized unmixing methods, the performance of SISAL is found better than MVSA with the resulting correlation values of 0.76 and 0.63, respectively, while also returning closer endmembers to the reference iron spectra. The performed research demonstrates the potential of hyperspectral applications on Sentinel-2 data to uncover the sub-pixel iron-induced spectral features in the visible region, proving compatible results between the spectral and laboratory analysis.