The thin-lipped grey mullet (Liza ramada) belongs to the Mugilidae family. Liza ramada mature females do not spawn without hormonal injection treatments in captivity. This study describes the application of human chronic gonadotrophic hormone (hcg), carp pituitary glands (cpg), Melatonin (MEL) and Mullet Brain extract (MBE) as an effective single-dose inducer of spawning in this species. A total of 40 wild mature female brooders (n = 10; 33.5 ± 3 cm total length, TL and 511 ± 20 g body weight, BW). The mature females were subjected into equal four groups as follow; the first group served as control (no hormonal treatment), while, the second and the third group received hormonal treatment of 4500 IU hcg in combined with 3 mg cpg per (hcg+cpg), and 3 mg melatonin (hcg+MEL). The last fish group received single hormonal treatment of 3 mg mullet brain (MB). The experiment results indicated significant increments (P > 0.01 or 0.001) on plasma estradiol (E2) and progesterone levels of adult female L. ramada fish received single injection dose from hcg in combined with cpg and MEL. Finally, the trial results suggest that mature L. ramada has recognized reproductive traits, adapts well to captive conditions, and can be easily induced to spawn by administering a single dose of human chronic gonadotrophic hormone (hcg) in combination with carp pituitary glands (cpg) and melatonin.