We firstly determined the CAC of the three conjugates. Pyrene has been widely used as the fluorescent probe for determining critical micelle concentration (CMC) and CAC with fluorescence spectrum. This method exploits the sensitivity of pyrene's emission spectrum to changes in its surrounding environment. Pyrene, a hydrophobic fluorescent probe, exhibits distinct fluorescence properties depending on whether it resides in a polar or nonpolar (hydrophobic) environment, such as the hydrophobic core of micelles. Specifically, the intensity ratio of the first and third peaks(I₁/I₃) in the emission spectrum of pyrene serves as an indicator of its microenvironment. In an aqueous solution, the I₁/I₃ ratio is higher due to the polar nature of water. As the concentration of a moleculereaches the CMC or CAC, thus pyrene was be concentrated within their hydrophobic cores of the aggregations, which lowers the I₁/I₃ ratio. Plotting the I₁/I₃ ratio against surfactant concentration allows for the identification of the CMC or CAC, represented by a sharp transition in the ratio .