The proposed study uses a special procedure to verify that an analyte assessment operation is sustainable and environmentally friendly. When the targeted analyte molecule (alogliptin drug, ALT) lacks chromophoric or fluorogenic groups, it cannot be analyzed without marking or structural amendment. The obeyed phenomenon (marking process) could be achieved via fluorescence probing or marking using tagging agents. An innovative reagent pair consisting of trioxosulfate ion and o-Phthaldialdehyde (OPD) is used in the described analytical approach. This reagent is used to build a novel fluorescent nucleus and can be detected using the developed fluorimetric approach. ALT's amino moiety could condense trioxosulfate/OPD. This study presents a new, facile, green, sustainable fluorometric assay of the ALT drug. The product's response is linear at 50.00–800.00 ng mL−1 concentration range (LOD = 15.74 and LOQ = 47.69 ng mL−1). The method validation was considered following the International Council for Harmonisation (ICH) recommendations. The planned probe was applied to the crude powder and dose forms, and excipients had no influence. The planned analytical procedure is identified as sustainable and white by the WAC (White Analytical Chemistry) indications following the Green/Red/Blue (RGB 12) pattern. Additionally, new tools (BAGI and GSST) were applied to the current work.