Laser trilobum (L.) Borkh., locally known as "Kefe kimyonu," is a perennial herb of the Umbelliferae family that is traditionally used in Turkey as a cumin substitute in culinary and folk medicinal applications. This study aimed to evaluate the chemical composition, antioxidant activity, and volatile compounds of ripe fruits (seeds) from L. trilobum plants grown in the Mersin/Adana region of the Taurus Mountains. Seven different seed samples were analyzed for their physicochemical and bioactive properties. The color values (L*, a*, b*) averaged 38.60 ± 2.65, 13.65 ± 3.47, and 1.34 ± 1.02, respectively. The seeds had a moisture content of 10.41 ± 1.16%, ash content of 10.00 ± 0.83%, crude fat content of 15.02 ± 2.55%, and crude protein content of 18.77 ± 1.32%. The total phenolic content was 7.91 ± 1.70 mg GAE/g, while the iron-reducing antioxidant power (FRAP) and DPPH radical scavenging capacity (IC₅₀) were 38.89 ± 20.97 mmol Fe²⁺/g and 298.24 ± 88.05 µg/mL, respectively. Volatile compound profiling via solid-phase microextraction (SPME) coupled with GC-MS revealed 20 major components, with limonene (47.99 ± 15.46%), α-pinene (19.19 ± 4.87%), and 1-cyclohexen-1-carboxaldehyde, 4-(1-methylhethenyl) (5.88 ± 2.46%) being the most abundant. This study represents one of the first comprehensive evaluations of L. trilobum seeds in terms of both chemical and aromatic composition. These findings suggest that L. trilobum seeds possess promising antioxidant properties and a rich volatile profile, supporting their potential use as a functional spice and as a natural ingredient in the cosmetic, fragrance, and pharmaceutical industries.