The pulverized manifestation of Pedalium murex seeds, excerpted by Soxhlets apparatus after treating with n-hexane. Oil sample was well scrutinized by EI-GC-MS, utilizing the full scan technique within mass ranges lies from 40-700 m/z. 73compounds were recognized among them, 63 compounds were identified and 10 were marked as unidentified (8, 22, 27, 43, 47, 61, 62, 64, 68 and 69). The method was executed by the conventional system of Mass spectroscopy and the data interpreted by considerable match factor ≥95 inspected by NIST library. Antidiabetic activity was carried out by Accu-Chek glucometer. Healthy albino mice were selected to perform antidiabetic activity of seed oil at 100mg/Kg and 200mg/Kg with a standard drug glibenclamide at 5mg/Kg. Antidiabetic activity was observed on 1st, 7th,14th, 21th,27th and 30th days. Statistical calculations and significant outcomes were obtained by One-way and Two-way analysis of variance, followed by Tuckey's test. The phytochemical n-hexadecanoic acid (19.53%) might be responsible for antidiabetic activity of the seed oil.