An "off-on" fluorescent probe, Nap-DNB, which is based on naphthimide, was designed and developed for the detection of biological selenols in vitro. We have adopted a combination of a low-pH detection environment and reaction sites that are more difficult to destroy to avoid the interference of a large number of biological thiols in biological samples. Nap-DNB can completely respond to selenocysteine within 15 mins, with a detection limit of 92 nM. Nap-DNB was successfully used for the detection of selenols in the serum, liver, and longissimus dorsi of selenium-enriched Tan sheep. Through comparison, we found that the detection of selenols by the Nap-DNB is similar to that by thioredoxin reductase and glutathione peroxidase in a commercial kit method. Nap-DNB can be used for the detection of selenols in selenium-enriched Tan sheep.
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