The aim of this study was to reveal the antidiabetic and antioxidant effects of ethanolic lyophilized extract of Achillea arabica flower extract against streptozotosine (STZ)-induced in diabetic rats and to determine its phytochemical content by liquid chromatography with tandem mass spectrometry (LC-MS/MS). After toxicity test, 35 female rats were divided into five groups. Control, diabetes mellitus (DM), A.arabica (400 mg/kg) extract, DM + A. arabica (400 mg/kg) extract and DM + Glibenclamide (2 mg/kg). It was determined that while diabetic rats treated A.arabica plant extract significantly decreased blood glucose level, serum glucose, HbA1c, liver and kidney damage biomarker levels, and malondialdehyde (MDA) content compared to the DM group, it caused fluctuations in antioxidant enzyme levels. According to LC-MS/MS results of A. arabica flower extract, quinic acid (2439.9 μg/g), cyranoside (858.4 μg/g), chlorogenic acid (698.7 μg/g), and cosmosiin (347.8 μg/g) were determined as major compounds, respectively. In addition, two new compounds were determined in this extract according to nuclear magnetic resonance (NMR) and Mass analyses and these compounds were named edremitine and achillosine, respectively. Thus, A.arabica flower extract has possible therapeutic effects to prevent high blood glucose level and oxidative stress caused by DM in liver and kidney via its high phenolic content.