Gelatin Speciation Using Analysis of Mass Spectrometry-based-Proteomics Datasets

Authentication of gelatin, an ingredient in food and drug products, is substantial with respect to some regulatory authorities and religion rules [1-3]. Muslim or Jewish people are concerned about gelatin derived from the porcine and fish without scales. The restrictions are released by regulatory authorities on the use of bovine gelatin in the case of the outbreak of Bovine Spongiform Encephalopathy (BSE) [4]. More specifically, the bovine skin-based gelatin is forbidden in Islam if the cattle are not slaughtered according to the religious rules. Even though different studies have been done for characterizing animal source origin of gelatin in the literature [3], there is not a method for tissue specification. Thus, developing an accurate and precise method is of interest for differentiation of bovine, porcine and fish gelatin and besides bovine skin and bone gelatin in food and drug products. The gelatin samples were prepared in the proper concentration and processed with overnight trypsinization [5-7]. An appropriate volume of trypsinized sample was loaded on HPLC-ESI-QTRAP mass spectrometer and injected twice to the instrument. The generated High-Performance Liquid Chromatography-Mass Spectrometry (HPLC/MS) data sets involve MS spectrum of tryptic gelatin peptides of the gelatin extract samples. Pattern recognition methods, PLS/DA and PCA methods [8, 9] are used for mining HPLC/MS data set to define the pattern and also, peptide markers for discrimination the extracted gelatins. The detected peptides markers are applied for multiple reaction monitoring (MRM) mass spectrometry in routine works. The method was then tested on the extracts of gummy bears, marshmallow as well as pharmaceutical capsules used for drug delivery. Results reveal that the proposed strategy is successful in the identifying of fish, bovine and porcine gelatin as well as bovine skin and bone gelatin in routine analysis of food and drug products. To best of our knowledge, a method for specifying bovine skin and bone gelatin is introduced for firstly in this study.