Hyptis suaveolensis is used in traditional medicine for the treatment of respiratory diseases, fever, skin diseases, pain and cramps. The essential oil has been reported as antifungal, antimicrobial, anti-inflamatory and also as an insecticide and a fungicide. The objectives of this work were to establish the effect of plant growth regulators, light emitting diode (LED) and light intensity on in vitro plantlet growth and volatile compound of Hyptis suaveolens. Nodal segments were obtained from mother plants as primary explants and placed on solidified half-strength MS medium supplemented with (BA × NAA). The explants were also placed at Photosynthetic photon flux (PFD) of 13, 28, 47 and 69 μmol m-2 s-1 and also in Light Emitting Diode (LED) blue; red; yellow; white; green; red: blue (4:10), (10:4) and (7:7). Analysis of the volatile fraction from leaf was done by Head-Space - GC/MS. Plant growth regulators tested for in vitro culture multiplication resulted in callus production after 4 weeks. Photon flux density and Light Emitting Diode (LED) affected plantlet growth and morphological characteristic of H. suaveolens. The production of volatile constituents was highly influenced by the type of growing conditions.