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Creating a Mass Spectral Reference Library for Oligosaccharides in Human Milk
Connie A Remoroza1, Tytus D Mak1, Maria Lorna A De Leoz1
1Mass Spectrometry Data Center, Biomolecular Measurement Division , National Institute of Standards and Technology , Gaithersburg , Maryland 20899-8362 , United States.
Analytical Chemistry
|July 4, 2018
Summary
A new mass spectral reference library for human milk oligosaccharides was developed. This library aids in identifying and characterizing complex oligosaccharides in biological fluids using advanced mass spectrometry techniques.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Glycomics
Background:
- Human milk oligosaccharides (HMOs) play crucial roles in infant health.
- Accurate identification and characterization of HMOs are challenging due to their structural complexity.
- Existing spectral libraries may not comprehensively cover the diversity of HMOs.
Purpose of the Study:
- To develop and provide a mass spectral reference library for oligosaccharides in human milk.
- To establish a searchable library of consensus spectra for annotated compounds.
- To enable the structural assignment of unknown oligosaccharides using a novel hybrid search algorithm.
Main Methods:
- Utilized hydrophilic interaction liquid chromatography electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS).
- Employed the NIST 17 Tandem MS Library and a newly developed hybrid search algorithm.
- Generated consensus reference spectra from NIST Standard Reference Material (SRM) 1953 (human milk).
- Applied Fourier transform ion trap (FT-IT) and higher energy dissociation (HCD) fragmentation techniques.
Main Results:
- Successfully identified and characterized 74 oligosaccharides, including fucosyllacto- and sialyllacto-oligosaccharides.
- Demonstrated the ability to distinguish between neutral and sialylated isomeric oligosaccharides based on retention times.
- Generated high-accuracy mass measurements for precursor and product ions.
Conclusions:
- The developed mass spectral reference library and hybrid search algorithm facilitate the structural assignment of complex oligosaccharides.
- This resource is applicable for analyzing oligosaccharides in various biological fluids beyond human milk.
- The library represents a significant advancement in glycomics research and metabolite identification.
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