Chlorite is often associated with hematite and tends to form microfine particles during grinding, making it challenging to recover and separate from hematite through flotation. This study innovatively applies an ester-based cationic polyacrylamide (CPAM) as a flocculant in a reverse flotation system using dodecylamine (DDA) as a collector, to flocculate and recover microfine-grained chlorite. The method achieves efficient separation between microfine chlorite and hematite. Laser particle size testing and optical microscopy show that CPAM significantly increases the apparent particle size of microfine chlorite, causing it to aggregate into flocs, while having a extremely weak effect on hematite flocculation. Further analysis using techniques like Zeta potential, contact angle measurement, Fourier-transform infrared (FTIR), and X-ray photoelectron spectroscopy (XPS) reveals CPAM's stronger adsorption affinity for chlorite compared to hematite. The ester groups in CPAM selectively form hydrogen bonds with chlorite's surface H atoms, enhancing its hydrophobicity. Particle-bubble attachment angle testing confirms that the flocculated chlorite flocs exhibit better floatability and higher efficiency in collision and adhesion with bubbles. Thus, CPAM serves as an efficient flocculant for reverse flotation separation of microfine chlorite and hematite.