Improved RP‐HPLC method for determination of bovine lactoferrin and its proteolytic degradation in simulated gastrointestinal fluids
作者： Xudong Yao , Craig Bunt , Jillian Cornish , Siew-Young Quek , Jingyuan Wen
DOI: 10.1002/BMC.2771
关键词:
摘要: The objective of this study was to qualitatively and quantitatively evaluate bovine lactoferrin (bLf) its stability using a rapid RP-HPLC method. bLf could be rapidly detected within 20 min quantitated at levels down 5 µg/mL, the equation linearity y = 86.10x + 178.31 with correlation coefficient (r(2)) 0.9997. Quantitative data obtained in present proved improved method sensitive accurate analytical tool for determination. proteolytic cleavage simulated human gastrointestinal fluids further analyzed by RP-HPLC, found follow pseudo-first-order kinetics. typical pepsin log(10) [A(t)]/[A(0)] -0.03x (r(2) 0.85), -0.01x 0.81) trypsin chymotrypsin combination. Pepsinolysis gastric fluid relatively fast half-life t(1/2) 23.1 min. digestion intestinal slower about 3-fold increase (69.3 min). After complete proteolysis bLf, small cleaved peptide fragments were fully separated identified RP-HPLC. indicated that validated able though monitoring derivatization products.
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