This work reports a facile and in situ method for synthesis of silver nanoparticles on the surface of eggshell membrane, which contains 3D interconnected porous networks and can be used for immobilization of metal nanoparticles. The as-prepared composites exhibit excellent SERS activities as substrates for 4-mercaptopyridine and Rhodamine 6G. The SERS measurements show detection limits of 5.0 x 10 (9) M for 4-mercaptopyridine and 1.0 x 10 (6) M for Rhodamine 6G on the prepared substrates, respectively. Furthermore, the uniformity and stability of this substrate have been studied and the results are satisfactory. The as-prepared substrate is potentially useful in practical SERS detection applications. (C) 2016 Elsevier B. V. All rights reserved.