![]() ![]() A LC/MS/MS system, a Nexera X2 UHPLC connected to LCMS-8060 which is same as that used for Halal testing method was employed to develop MRM based method for detection and differentiation of bovine and porcine-specific peptide markers. The details of the UHPLC and MS/MS parameters are shown in Table 1. Analytical conditions for detection and differentiation of bovine and porcinespecific peptide markers on LCMS-8060Īeris Peptide 1.7 μm XB-C18 100Å (150 mm x 2.1 mm I.D.) Protocol to obtain gelatin references from raw meat materials Figure 2 shows the details of the procedure.įigure 2. These gelatin reference materials can last for months at -20oC. In this report, in-house gelatin references derived from raw materials such as beef tendon and pork skin are used. ![]() However, the stability of those CRM remained unverified as bovine and porcine-specific markers were detected at trace levels in both standards (data not shown). Experimental protocol for gelatin extraction, digestion and LC/MS/MS analysisĬommercial certified reference materials (CRM) of gelatins were obtained and used to confirm detection of peptide markers. Then, the capsule shell is cut using clean scissor and placed into extraction tube.įigure 1. The capsule shell of pharmaceutical product is rinsed with milli-Q-water to remove any content. Because low concentration of ammonium bicarbonate is compatible to LC/MS/MS, the liquid sample (supernatant) is subjected directly to analysis without dilution. However, before injection to LC/MS/MS, centrifugation of sample is applied to segregate the digested peptides from insoluble solid extract. The experimental protocol is comprised of three steps including gelatin extraction, tryptic digestion and LC/MS/MS analysis (Figure 1). Preparation of gelatin samples and referencesĮxtraction of gelatins from samples is relatively undemanding due to their high solubility in water, as compared to the extraction of proteins for specific peptide markers. The established LC/MS/MS method was proven to be a robust approach for source authentication of gelatin of processed food and pharmaceutical capsules. The specificity of more than 40 signature peptides as marker candidates was tested using in-house gelatin references due to the similarity in their sequences and properties. ![]() As a part of application news series to support Halal authentication testing method based upon targeted proteomic approach, a practical method by using sensitive LC/MS/MS was developed to pinpoint species-specific peptide markers for most widely used gelatins, bovine and porcine, as well as to determine gelatin source in commercial food and pharmaceutical products. Accurate food authentication and labelling are requisite for consumers’ assurance towards the products. Furthermore, there is a concern of animal-based gelatin could be a media for zoonotic disease risks. However, general approval of animal-based gelatin is restricted due to incongruity with several religious customs and individual dietary preference which require the gelatin to be free from porcine, bovine or any animals-byproducts. Commercial gelatins are often manufactured from bovine and porcine materials (skin, bone and hide etc.) because of the practicability and cost-effectiveness. Gelatin is a mixture of polypeptides derived from hydrolysis of collagens of animal origins and has been widely utilized in food, cosmetic, and pharmaceutical industries. Halal Authentification Analysis / LCMS-8060Īpplication Development & Support Centre, Shimadzu (Asia Pacific) Pte Ltd, Singapore ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |