Journal
ANALYTICAL CHEMISTRY
Volume 94, Issue 7, Pages 3142-3149Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c04349
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Funding
- Global Water Futures Project of the Canada First Research Excellence Fund (CFREF)
- Natural Sciences and Engineering Research Council of Canada (NSERC)
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In this study, four high-quality aptamers for caffeine were isolated and their binding patterns were explored. Additionally, a fluorescent sensor was designed to detect caffeine concentration in different samples, achieving a low detection limit even in complex body fluids.
With the growing consumption of caffeine-containing beverages, detection of caffeine has become an important biomedical, bioanalytical, and environmental topic. We herein isolated four high-quality aptamers for caffeine with dissociation constants ranging from 2.2 to 14.6 mu M as characterized using isothermal titration calorimetry. Different binding patterns were obtained for the three single demethylated analogues: theobromine, theophylline, and paraxanthine, highlighting the effect of the molecular symmetry of the arrangement of the three methyl groups in caffeine. A structure-switching fluorescent sensor was designed showing a detection limit of 1.2 mu M 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 mu M caffeine was achieved. With the four aptamer sensors forming an array, caffeine and the three analogues were well separated from nine other closely related molecules.
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