There is an urgent need for rapid and reliable methods able to detect melamine in animal feed. In this study, a quick, simple, and sensitive method for the determination of melamine content in animal feed was developed using surface-enhanced Raman spectroscopy on fabricated Ag nanorod (AgNR) array substrates with a one-step sample extraction procedure. The AgNR array substrates washed by HNO3 solvent (10-7 M) and methanol and showed the good stability within 6 months. The Raman shift at △ν = 682 cm-1 was used as the characteristic melamine peak in the calculations. Sufficient linearity was obtained in the 2-200 μg·g-1 range (R2 = 0.926). The limits of detection and quantification were 0.9 and 2 μg·g-1, respectively. The recovery rates were 89.7-93.3%, with coefficients of variation below 2.02%. The method showed good accuracy compared with the tradition GC-MS analysis. This new protocol only need 2 min to fininsh the detection which could be developed for rapid onsite screening of melamine contamination in quality control and market surveillance applications.