Industrialization is serious for changing the environment and natural water composition, especially near cities and manufacturing areas. Logically, the new ultrasensitive technology for precise control of the quality and quantity of water sources is needed. Herein, an innovative method of polarization fluorescence analysis (FPA) was developed to measure the concentration of heavy metals in water. The approach was successfully applied for precise tests with reduced analysis time and increased measurement efficiency among laboratory methods. Based on this work, the investigations established the new type of carbon quantum dots (CQDs) with controllable fluorescence properties and functionalized amino—groups, which is appropriate for FPA. The parameters of one and two-step microwave synthesis routes are adjusted wavelength and fluorescence intensity of CQDs. Finally, under optimized conditions, the FPA is showed the detection of copper (2+) cations in water samples below European Union standard (2 mg/L). Moreover, in comparison with fluorescence quenching, polarization fluorescence is proved as a convenient, simple, and rapid test method for effective water safety analysis.