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Nanoscale zero-valent iron particles (NZVI) produced by using green tea (GT) extract as a reductant can remove Cr(VI) from water effectively, which can be utilized in groundwater remediation. In order to define the reaction mechanism and removal effect in the aquifer, in this study, GT-NZVI particles were prepared and measured by some characterization methods to define their surface performance, and then batch and one-dimensional experiments were carried out to reveal the reaction properties of GT-NZVI and Cr(VI) in groundwater. The results showed that the prepared GT-NZVI particles were regular spherical with a diameter of 10–20 nm, which could disperse in water stably. The main component of GT-NZVI was α-Fe with superficial polyphenols as a stabilizer. GT-NZVI suspension had good ability to reduce the Cr(VI) to Cr(III) in water. When the concentration of GT-NZVI was 1 g/L, the removal efficiency of Cr(VI) with an initial concentration of 100 mg/L reached 92.8% in 1 h reaction. In column tests, GT-NZVI passed through the natural sand column successfully with an average outflow percentage of 71.2%. The simulated in-situ reaction zone (IRZ) with GT-NZVI was used to remediate Cr(VI) contaminated groundwater. The outflow concentration of Cr(VI) kept in 0.14–0.32 mg/L corresponding to the outflow rate below 0.32% within 15 days, and the removal efficiency of Cr(VI) by IRZ with GT-NZVI decreased with the increase of aquifer medium particle size, groundwater flow rate and ionic strength. Most of Cr(III) as reduzate was adsorbed or immobilized on the surface or in the lattice of GT-NZVI, which indicated effective immobilization for chromium.