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[Display omitted] • A unique SERS substrate processed by one-step femtosecond laser ablation in Argon. • S-Ag-Ar can detect 10-8mol/L R6G with enhancement factor up to 5.3 × 106. • S-Ag-Ar has high uniformity with relative standard deviation less than 11.97%. • S-Ag-Ar can be stored for more than seven months and shows good universality. • This unique SERS substrate demonstrates good potentials for food safety evaluation. Surface Enhanced Raman Scattering (SERS) is a reliable tool for detection of food additives due to its advantages including rapid detection, non-destructive and excellent sensitivity. Currently, the commercial SERS substrates are not yet satisfactory referring to cost, sensitivity, uniformity and stability. Herein, we report a unique SERS substrate processed by one-step femtosecond laser ablation in Argon. The substrate shows high sensitivity with limit of detection up to 10-8mol/L for Rhodamine 6G (R6G) and enhancement factor up to 5.3 × 106 due to laser induced more plentiful nanostructures and less oxidation in Argon, in addition to high uniformity with relative standard deviation less than 11.97% and long storage time more than seven months. This SERS substrate is capable of detecting the common food additives including malachite green (1 ppm), crystal violet (10-7mol/L), Sudan I (1 ppm), melamin (1 ppm), brilliant blue (1 ppm) and tartrazine (1 ppm). This unique Ag SERS substrate demonstrates good potentials for food safety evaluation. [ABSTRACT FROM AUTHOR]