Improved RP-HPLC method for determination of bovine lactoferrin and its proteolytic degradation in simulated gastrointestinal fluids

Show simple item record Yao, Xudong en Bunt, C en Cornish, Jillian en Quek, Siew-Young en Wen, Jingyuan en 2016-10-05T23:18:25Z en 2013-02 en
dc.identifier.citation Biomedical Chromatography, 2013, 27 (2), pp. 197 - 202 en
dc.identifier.issn 0269-3879 en
dc.identifier.uri en
dc.description.abstract The objective of this study was to qualitatively and quantitatively evaluate bovine lactoferrin (bLf) and its stability using a rapid RP-HPLC method. bLf could be rapidly detected within 20 min and quantitated at levels down to 5 µg/mL, and the equation of linearity was y = 86.10x + 178.31 with the correlation coefficient (r(2) ) 0.9997. Quantitative data obtained in the present study proved the improved RP-HPLC method to be a sensitive and accurate analytical tool for bLf determination. The proteolytic cleavage of bLf in simulated human gastrointestinal fluids was further analyzed by RP-HPLC, and found to follow pseudo-first-order kinetics. The typical equation obtained by pepsin was log(10) [A(t) ]/[A(0) ] = -0.03x (r(2)  = 0.85), and log(10) [A(t) ]/[A(0) ] = -0.01x (r(2)  = 0.81) for trypsin and chymotrypsin combination. Pepsinolysis of bLf in simulated gastric fluid was relatively fast with the half-life t(1/2) 23.1 min. The digestion of bLf in simulated intestinal fluid was slower with about a 3-fold increase in half-life (69.3 min). After the complete proteolysis of bLf, small cleaved peptide fragments were fully separated and identified by RP-HPLC. The proteolytic study indicated that this validated RP-HPLC was able to evaluate bLf stability though monitoring the derivatization products. en
dc.description.uri en
dc.publisher Wiley en
dc.relation.ispartofseries Biomedical Chromatography en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. Details obtained from en
dc.rights.uri en
dc.title Improved RP-HPLC method for determination of bovine lactoferrin and its proteolytic degradation in simulated gastrointestinal fluids en
dc.type Journal Article en
dc.identifier.doi 10.1002/bmc.2771 en
pubs.issue 2 en
pubs.begin-page 197 en
pubs.volume 27 en
dc.rights.holder Copyright: Wiley en
dc.identifier.pmid 22684820 en en
pubs.end-page 202 en
pubs.publication-status Published en
dc.rights.accessrights en
pubs.subtype Article en
pubs.elements-id 371952 en Medical and Health Sciences en Pharmacy en School of Medicine en Medicine Department en Science en Chemistry en Science Research en Maurice Wilkins Centre (2010-2014) en
dc.identifier.eissn 1099-0801 en
pubs.record-created-at-source-date 2012-08-21 en
pubs.dimensions-id 22684820 en

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