Another chiral crystal of 1-[4-(dimethylamino)benzylidenel-4-phenylthiosemicarbazide (DMB) was obtained, and its absolute structure was determined by X-ray single crystal diffraction and the solid circular dichroism (CD) spectrum. The crystal belongs to orthorhombic system with space group P2(1)2(1)2(1) and a = 0.7870(2), b = 1. 1560(2), c = 1.6668(3) nm(3), V = 1.5164(5) nm, Z= 4, D-c=1.307 g/cm(3), F(000)=632, mu=0.213 mm(-1), the final R=0.0409 and wR=0.1061 for 2577 observed reflections [I > 2 sigma(I)]. The absolute structure can be determined well with the Flack parameter 0.00(9). The crystal structure and CD spectrum show that an enantiomer of the compound packs into the chiral crystal