Types of forage and the altitude it grows affect the quality of milk’s fatty acids (FA) profile. Thus, this study aimed to analyze the chemical and FA quality of elephant grass (EG) and king grass (KG) growth from different altitudes. The grass samples were collected from Pangalengan District, the highland, and Dramaga District, the lowland. Grass lipid extraction and FA methylation were done according to the AOAC method. FA was quantified using gas chromatography (GC-7820A), CP-Sil 88 fused-silica capillary column, and authentic standard (Supelco 37 Component FAME-Mix). The nutrient content was analyzed using FT-NIR Spectrometer Solids Cell, except for EE used the AOAC method. This study showed EE content was significantly different between species, with the higher content found in EG. The C17:0 was significantly different between altitudes. The SFA, UFA, and PUFA were significantly different between species. The EG had higher UFA and PUFA and lower SFA. It showed that the higher EE content was related to the higher FAs yield, followed by higher UFA and PUFA in EG than in KG. It was concluded that the grass species had a more significant effect on chemical and FAs profiles than the altitude.