Consumption of food products derived from porcine sources is strictly prohibited in Islam. Gelatin,
mostly derived from bovine and porcine sources, has many applications in the food and pharmaceutical
industries. To ensure that food products comply with halal regulations, development of valid and reliable
analytical methods is very much required. In this study, a species-specific polymerase chain reaction
(PCR) assay using conserved regions of mitochondrial DNA (cytochrome b gene) was performed to evaluate
the halal authenticity of gelatin. After isolation of DNA from gelatin powders with known origin, conventional
PCR using species-specific primers was carried out on the extracted DNA. The amplified
expected PCR products of 212 and 271 bp were observed for porcine and bovine gelatin, respectively.
The sensitivity of the method was tested on binary gelatin mixtures containing 0.1%, 1%, 10%, and
100% (w/w) of porcine gelatin within bovine gelatin and vice versa. Although most of the DNA is degraded
due to the severe processing steps of gelatin production, the minimum level of 0.1% w/w of both porcine
and bovine gelatin was detected. Moreover, eight food products labeled as containing bovine gelatin and
eight capsule shells were subjected to PCR examination. The results showed that all samples contained
bovine gelatin, and the absence of porcine gelatin was verified. This method of species authenticity is very
useful to verify whether gelatin and gelatin-containing food products are derived from halal ingredients.