Identification of Lymphoid Tissues in Pork by an Inflammation and Immunity-related Metabolite Quantification Combined with Chemometrics
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Abstract:
Identification of pork with lymphoid tissues based on quantitative analysis of inflammation and immunity-related metabolites through ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was studied. Samples were extracted with 80% acetonitrile-water and degreased with n-hexane. The separation was carried out using a BEH C18 column (2.1 mm×100 mm, 1.7 μm), with 5 mmol/L ammonium acetate solution-acetonitrile as the mobile phase. Gradient elution was conducted for 25 min, and positive and negative ions were determined simultaneously. A quantitative method for 38 inflammation and immunity-related metabolites based on UPLC-MS/MS was established to quantitatively examine these metabolites in ordinary pork and pork with lymphoid tissues, and the identification model for the latter was constructed by chemometrics. The results reveal a good linear relationship between the target compounds in the concentration range of interest. The recoveries of low concentration spiked samples range from 50.3% to 118.8%, and the recoveries of high concentration spiked samples range from 57.0% to 127.0%, with RSDs below 20%. Based on the PCA and OPLS-DA models, the pork samples with lymphoid tissues and ordinary pork samples can be well identified, and the samples with lymphoid tissue proportions more than 20% can be distinguished from ordinary samples. The R2 and Q2 were 0.954 and 0.858, respectively, and the model showed no over-fitting and good prediction performance. The contents of inflammation and immunity-related metabolites combined with chemometrics can effectively distinguish pork samples with lymphoid tissues from ordinary pork samples, providing a reliable method for the identification of pork with lymphoid tissues.
