Berberine (BBR) is widely used and of significant importance in medical field, and its quantitative analysis is of great value in guiding clinical treatment. There still be some shortcomings remaining in the determination of berberine by traditional methods, such as less sensitivity, narrow response range, or high testing cost. In this study, we found that hematoporphyrin monomethyl ether (HMME) with negative groups had a high fluorescence quenching effect on Tetra (trimethylammonia) aluminum phthalocyanine (TTMAAlPc), a cationic phthalocyanine compound emitting strong red fluorescence, due to the formation of an almost non-fluorescent ion associate. Interestingly, it was found that the fluorescence of TTMAAlPc-HMME associate re-emitted in the presence of berberine. The recovery of fluorescence of TTMAAlPc was positively correlated with the concentration of berberine. The mechanism of the above-mentioned phenomena was discussed. We believed that the high-efficiency fluorescence quenching effect of HMME on TTMAAlPc can be attributed to the formation of associate consisting of hematoporphyrin monomethyl ether and the cationic aluminium phthalocyanine (HMME-TTMAAlPc) through supramolecular interactions including planar hydrophobic interaction, π-bond stacking, electrostatic interaction and hydrogen bonding mediated by water molecules. The fluorescence recovery was due to the strong competitive binding to release the cationic aluminum phthalocyanine from the associate by berberine with a molecular structure matches TTMAAlPc. A new fluorescence-enhanced quantitative analysis method for berberine was established based on the above-findings. The factors including pH, sorts of buffer, reaction temperature, usages of HMME and TTMAAlPc and reaction time that affected the determination were investigated. Under the optimum conditions, the equation of calibration curve was If=12.85c+66.30 with a correlation coefficient (R) of 0.997 2, the linear range was 0.3~69.0 mg·L-1, and the detection limit was 0.01~5.0 mg·L-1. The study indicated that this method bears the merits of strong specificity, simple operation and low cost. The established method has been applied to the determination of practical samples with satisfactory results. This study not only presents a new strategy for the construction of phthalocyanine-based optical probes, but also opens up the application of red-emitting phthalocyanines in the field of drug analysis.