Many pharmaceuticals include highly potent active pharmaceutical ingredients (API), which only require a small dosage to obtain the desired therapeutic effect. This leads to a challenge for quantification of the API using process analytical technology, since the standard nondestructive measurement technique, near-infrared spectroscopy, is not able to quantify below 1% (weight/weight (w/w)) API content. In formulations with more than one API, this challenge is further increased. The purpose of this study is to scrutinize the potential of fluorescence spectroscopy for the simultaneous quantification of two APIs: flupentixol (FLU) in low dosage (0.208-0.625% w/w free base) and melitracen (MEL) (4.17-12.5% w/w free base) in a tablet formulation. Despite internal quenching between the ingredients and the two APIs, this paper demonstrates that it is possible to establish calibrations using partial least squares (PLS) regression on unfolded fluorescence landscapes with a root mean square error of prediction and relative error of 0.038% (w/w) and 9.1%, for FLU and 0.344% (w/w) and 4.1% for MEL, respectively.