The search for natural antioxidants as alternatives to synthetic products is growing. Microalgae have emerged as a source of natural antioxidants with significant and diverse health-promoting properties. In this study, the effects of hot-water extraction conditions on total polyphenol compounds (TPC) production were investigated for lipid extracted microalgae (LEA). In order to enhance the polyphenol productivity, the extraction variables including solvents, temperature, time and ethanol concentration were optimized. The results showed hot-water extraction provided a higher extraction efficiency than the organic solvents and extraction at high temperatures showed a better extraction efficiency. While hot-water extract showed a higher extraction efficiency compared to 98% ethanol extraction, the mixture of water and ethanol (40:60 v/v) showed the highest production of polyphenols. The maximum polyphenols of 3.35 mg GAE (gallic acid equivalent)/g DM were obtained at the optimized extraction time of 10 min, 100 ℃ and 40% ethanol, respectively. Although Tetraselmis KCTC 12236BP was preprocessed by hexane to remove lipid for bio-diesel production, the results showed LEA contains relatively high level of polyphenols compared to untreated microalgae which can be used in the production of value-added materials. The predictions obtained from the developed Peleg’s model were compared with the experimental data under the same operating conditions. The predicted and experimental data were consistent, indicating the reliability of the model.