A system is proposed for generation of security key using R-R interval feature of electrocardiogram (ECG) signals as an input. The system provides a mechanism for verification and identification by generating a security key corresponding to an individual. The system comprises of two independent stages: registration and authentication. Biometric security key, created in the registration stage has been generated using Hamming Distance and the extended version of the triple DES algorithm. Biometric security key generation, verification, authentication and performance of the biometric security key have been assessed using R-R interval of ECG signals taken from the standard MIT-BIH database. The performance of the proposed biometric security key has been tested for the parameters like FAR, FRR, GAR, HTER and accuracy. The simulation results for 64-bit, 128-bit and 256-bit biometric security key indicate that the performance of the proposed biometric security key is reasonably good for a security system.