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Chromatography 2016
September 21-23, 2016
Volume 7, Issue 5(Suppl) J Chromatogr Sep Tech 2016
ISSN: 2157-7064 JCGST, an open access journal
conferenceseries .com
September 21-23, 2016 Amsterdam, Netherlands
World Congress on Chromatography
Analysis of natural crosslinks by liquid chromatography mass spectrometry using a silica hydride column
Rafea M Naffa1, Meekyung Ahn2 , Richard Haverkamp1 and Gillian E Norris1
1Massey University, New Zealand
2Leather and Shoe Research Association, New Zealand
The natural crosslinks between tropocollagenmolecules in skin contribute to its physical properties of strength and flexibility.
Despite advances in analytical techniques such as mass spectrometry, the methods used for their analysis and quantitation
date back to the 70s through to the 90s, are challenging and time consuming. As a result, it is difficult to obtain standards to verify
analyses of these compounds. We have developed methods to isolate highly pure, natural crosslinks from skin, and have verified
their structures using mass spectrometry and NMR. Fragmentation studies of these crosslinks will enable the development of
a method for label-free quantitation in skin hydrolysates. We have also developed a novel chromatographic analytical method
for the simultaneous determination of natural crosslinks. Seven crosslinks were separated on Cogent Diamond Hydride HPLC
column using isocratic and gradient conditions then detected by mass spectrometry without derivatization. Total run time of
less than 10 minutes was achieved under isocratic conditions using water and acetonitrile. To the best of our knowledge, this
is the first method in which histidinolysinonorleucine (HHL) and histidinohydroxymerodesmosine (HHMD) were separated
and identified by the mass spectrometry. This technique was applied on skin, elastin and cartilage in which strong evidence
suggested the presence of undocumented crosslinks. The developed method will be widely used for quantitative and qualitative
analysis of natural crosslinks in biological samples as well as characterization of new crosslinks.
Biography
Rafea M Naffa is pursuing his PhD from Massey University, New Zealand. He has worked as Lecturer at University of Sharjah from a period of 2007-2014. He
completed his Master’s in Chemistry from Hashemite University. His research interest includes Chemistry, Bio-Analytical Chemistry and Leather Chemistry.
Rafea M Naffa et al., J Chromatogr Sep Tech 2016, 7:5(Suppl)