Detection of Type I and III collagen in porcine acellular matrix using HPLC–MS

Acellular matrix (ACM) has been widely used as a biomaterial. As the main component of ACM, collagen type and content show influence on the material properties. In this research, the collagen in ACM from different tissues of pig were determined by detection of marker peptides. The marker peptides of Type I and III collagen were identified from the digested collagen standards using ions trap mass spectrometry (LCQ). The relationship between the abundance of marker peptide and collagen concentration was established using triple quadrupole mass spectrometer (TSQ). The contents of Type I and III collagen in ACM from different tissues were determined. The method was further verified by hydroxyproline determination. The results showed that, the sum of Type I and III collagen contents in the ACM from small intestinal submucosa, dermis and Achilles tendon of pig were about 87.59, 81.41 and 61.13%, respectively, which were close to the total collagen contents in these tissues. The results proved that this method could quantitatively detect the collagen with different types in the ACM of various tissues.