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Summary The gelatin sources have become a controversial issue with regard to religious and health concern. Thus, the aims of this study were to develop and evaluate the efficiency of polyclonal antibodies against peptide immunogen of collagen α2 (I) chain for determination of gelatin sources in confectionery products by competitive indirect enzyme‐linked immunosorbent assay (ELISA). Collagen α2 (I) chain protein showed resistance against heat treatment and detectable in certain commercial products when analysed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE). The established ELISA exhibited low cross‐reactivity to fish and chicken gelatin. The IC50 value was 0.39 μg mL−1, and the limit of detection (IC10) was 0.05 μg mL−1. There were no false‐positive results from forty‐eight commercially processed products. The present method is useful for determination of gelatin in confectionery products.

a b s t r a c tCompetitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for rapid detection of porcine gelatin in edible bird's nest (EBN). Three ELISAs were developed by using polyclonal rabbit antibodies against porcine species-specific amino acid sequences of collagen a2 (I) chain (pAb1 and pAb2) and a1 (I) chain (pAb3). The limit of detection (IC 15 ) of the three ELISAs was 0.033, 0.082 and 0.052 mg/mL respectively. The median inhibitory concentration (IC 50 ) of pAb1, pAb2 and pAb3 was 0.265, 0.394 and 0.228 mg/mL respectively, as well as able to recognise porcine and bovine gelatins. pAb1showed slight cross-reactivity with cave nest and egg white, while pAb2 exhibited slight cross-reactivity with blood cave nest and egg white. No cross-reactivity was observed with EBNs and egg white for pAb3. The recoveries of porcine gelatin spiked EBNs were in the range of 62.8e125.4% with intra-and inter-day coefficient of variants (CVs) of 2.9e5.4% and 4.7e9.6% respectively when using pAb3. Taking into account all abovementioned factors, pAb3 appeared sufficient for EBN authentication.