Abstract: Quantitative analysis of fibronectin in porcine small intestinal submucosa(p-SIS) acellular matrix was investigated using batch enzymatic digest and high performance liquid chromatography mass spectrometry(HPLC-MS). The results indicated that batch enzymatic digest could improve the hydrolysis efficiency compared to static enzymatic hydrolysis. The hydrolysis time decreased to 6 hours from 96 hours, and the enzymatic hydrolysis rate amounted to 95%. Marker peptide, SSPVVIDASTAIDAPSNLR, was detected from the tryptic digest of FN. Based on the peak areas of marker peptide from FN standards with different concentration, the FN content in VIDASISTM was determined as 0.43%. Amount of FN in Biodesign was determined as 0.17%. The method precision(RSD, 1.31%) and the deviation were carried out in the methodology experiments. The results indicate the established quantitative method is effective for detecting FN content in acellular matrix.

Key words: porcine small intestinal submucosa, extracellular matrix, fibronectin, marker peptide, LC-MS