Selection of Aptamers for Sensing Caffeine and Discrimination of Its Three Single Demethylated Analogues

With the growing consumption of caffeine-containing beverages, detection of caffeine has become an important biomedical, bioanalytical, and environmental topic. Four high-quality aptamers for caffeine were isolated and were carefully characterized. A structure-switching fluorescent sensor was designed showing a detection limit of 1.2 μM caffeine, which reflected the labeled caffeine concentration within 6.1% difference for eight commercial beverages. In 20% human serum, a detection limit of 4.0 μM caffeine was achieved. In addition, these four fluorescent aptamer sensors can form a sensing array, caffeine and the three analogues were well separated from nine other closely related molecules.