Abstract Authentication of gelatin, an ingredient in food and drug products, is of interest with respect to some regulations and religion rules. Determination of the gelatin origin is particularly challenging because of the similarity in amino acid sequences of collagen types in different species. In this study, Polymerase Chain Reaction (PCR) technique using species-specific primers and also chemometrics analysis of mass spectral data were investigated to differentiate bovine, porcine and fish source of gelatin in commercial pure gelatin and gelatin-containing food and drug products. The specific PCR primer set was optimized using real-time PCR approach to routinely determine the gelatin source. Real-time PCR is sensitive to identify gelatin origin from traces of species-specific DNA. However, source determination is impossible when DNA is denatured or removed during production process. Alongside, chemometrics methods (PLS/DA and PCA) were used for mining LC/MS data sets to obtain patterns for discrimination of gelatin origins. LC/MS data sets involve MS spectrum of tryptic gelatin peptides generated from gelatin samples. Using PLS/DA method, a discrimination model is developed for identifying origin of the extracted gelatin. The method is practical to determine the halal authenticity of gelatin in the pure and processed products.

中文翻译：

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使用实时 PCR 进行明胶物种形成和基于质谱的蛋白质组学数据集分析

摘要 明胶是食品和药品中的一种成分，在某些法规和宗教规则方面受到关注。由于不同物种胶原蛋白氨基酸序列的相似性，明胶来源的确定尤其具有挑战性。在这项研究中，研究了使用物种特异性引物的聚合酶链反应 (PCR) 技术以及质谱数据的化学计量学分析，以区分商业纯明胶和含明胶的食品和药品中的牛、猪和鱼​​来源的明胶。使用实时 PCR 方法优化特定 PCR 引物组，以常规确定明胶来源。实时 PCR 对从物种特异性 DNA 的痕迹中识别明胶来源很敏感。然而，当 DNA 在生产过程中变性或去除时，无法确定来源。此外，还使用化学计量学方法（PLS/DA 和 PCA）挖掘 LC/MS 数据集，以获得识别明胶来源的模式。LC/MS 数据集涉及从明胶样品中生成的胰蛋白酶明胶肽的 MS 谱。使用 PLS/DA 方法，开发了用于识别提取明胶来源的判别模型。该方法适用于确定纯品和加工品中明胶的清真真伪。开发了一种鉴别模型，用于识别提取明胶的来源。该方法适用于确定纯品和加工品中明胶的清真真伪。开发了一种鉴别模型，用于识别提取明胶的来源。该方法适用于确定纯品和加工品中明胶的清真真伪。