New chiral bisboronic receptors based on pyrene-excimer were synthesized and applied for the determination of absolute configuration and enantiomeric composition of tartaric acid. The distinction was visualized by portable UV lamp in certain concentration range. There were three ways for the sensors to enantioselectively recognize tartaric acid, including the fluorescence spectrum, UV–vis spectrum, and CD spectrum. The enantiomeric excess of tartaric acid was measured quantitatively by using fluorescence analytical technique. Compared with the previous chiral boronic acid sensors, sensor 1 showed weaker background fluorescence and excellent chiral recognition ability of D/L-tartaric acid throughout the whole pH range.