A simple spectrophotometric method for measuring high concentration of chloroform in water was developed based on the Reimer-Tiemann reaction. The reaction between chloroform and resorcinol under alkaline conditions has the limit of detection (LOD) and limit of quantification (LOQ) values at 1.28 mg/mL and 3.88 mg/mL, respectively. The experimental and response surface methodology (RSM) proposed the optimal conditions for the removal of chloroform (RoC) using tannic acid as a polyphenolic co-activator and spectrophotometric method for chloroform detection. The acceptable dechlorination process was suggested as persulfate (0.1–0.11 M), NaOH (0.6–0.8 M), and tannic acid (1.8 g/L) to obtain the yield of RoC from 76.14 to 77.90%. In situ experiments were then designed for RoC from the laboratory’s aqueous and organic solvent waste. After being treated with the system mentioned above, the yields of RoC were around 70%. In addition, a critical aspect of the innovation process was replacing pure tannic acid with other natural agents, which are more economically efficient and eco-safety. The EtOH extract from Elaeocarpus hygrophilus leaves (EEL) was suggested as a natural source of polyphenols, which could co-activate persulfate, with the yield of RoC rising to 63.29%. The findings of this study provide a novel strategy for chloroform degradation from laboratory wastewater using polyphenols.