A simple fluorescence method is described for measuring rutin dependent on the nitogen-doped carbon dots (NCDs) prepared via simple pyrolysis method from chicken feet biowaste. The as-fabricated NCDs have unique advantages including cost-effectiveness and high quantum yield (42.9 %). The as-prepared NCDs explore an optimal emission band at 430 nm following exciting NCDs at 330 nm. Addition of rutin to blue-emissive NCDs quenched their fluorescence emission by inner-filtration effect (IFE) and static quenching. Under optimized conditions, the fluorescence responses increased as the rutin amount was raised from 100 to 1000 nmol/L with 5.3 nmol/L as a detection limit (S/N = 3). The probe selectivity was improved by adding bovine serum albumin (BSA), which binds other structurally-related compounds (other flavonoids). The as-synthesized NCDs exhibited some advantages towards rutin detection such as: lower LOD value, satisfactorily reproducibility, simplicity, rapidity, selectivity, and stability. The sensing probe was efficiently utilized for determining rutin in different real samples with acceptable results. The sensor offers an efficient biowaste-based approach for the determination of (bio) molecules.