Detection method of capsaicinoids such as capsaicin, dihydrocapsaicin and nordihydrocapsaicin was developed using anti-homovalic acid-Concholepas concholepas hemocyanin (CCH) polyclonal antibodies and surface plasmon resonance (SPR) sensor with indirect competitive assay method. Extraction procedure of capsaicinoids from Capsicum fruit was also developed. As a result, limits of detection for capsaicin, dihidrocapsaicin and nordihydrocapsaicin were 27 ppb, 35 ppb and 50 ppb, respectively. The reduction of responses in SPR measurements with indirect competitive method for recovered samples of sweet peppers such as Bell pepper and Shishitougarashi were not observed. Nine kinds of hot peppers (eg. Habanero, Trinidad scorpion), which have different Scovile heat unit (SHU), were also tested. The reductions of sensor responses for the hot peppers were observed. It is likely that samples extracted from Capsicum fruit of the hot peppers contained capsaicinoids. It was found that the responses of the SPR immunosensor depended on SHU of each hot pepper.