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Label-Free Absolute Quantitation of Oligosaccharides Using Multiple Reaction Monitoring
[Image: see text] An absolute quantitation method for measuring free human milk oligosaccharides (HMOs) in milk samples was developed using multiple reaction monitoring (MRM). To obtain the best sensitivity, the instrument conditions were optimized to reduce the source and postsource fragmentation p...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical
Society
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3983013/ https://www.ncbi.nlm.nih.gov/pubmed/24502421 http://dx.doi.org/10.1021/ac404006z |
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author | Hong, Qiuting Ruhaak, L. Renee Totten, Sarah M. Smilowitz, Jennifer T. German, J. Bruce Lebrilla, Carlito B. |
author_facet | Hong, Qiuting Ruhaak, L. Renee Totten, Sarah M. Smilowitz, Jennifer T. German, J. Bruce Lebrilla, Carlito B. |
author_sort | Hong, Qiuting |
collection | PubMed |
description | [Image: see text] An absolute quantitation method for measuring free human milk oligosaccharides (HMOs) in milk samples was developed using multiple reaction monitoring (MRM). To obtain the best sensitivity, the instrument conditions were optimized to reduce the source and postsource fragmentation prior to the quadrupole transmission. Fragmentation spectra of HMOs using collision-induced dissociation were studied to obtain the best characteristic fragments. At least two MRM transitions were used to quantify and identify each structure in the same run. The fragment ions corresponded to the production of singly charged mono-, di-, and trisaccharide fragments. The sensitivity and accuracy of the quantitation using MRM were determined, with the detection limit in the femtomole level and the calibration range spanning over 5 orders of magnitude. Seven commercial HMO standards were used to create calibration curves and were used to determine a universal response for all HMOs. The universal response factor was used to estimate absolute amounts of other structures and the total oligosaccharide content in milk. The quantitation method was applied to 20 human milk samples to determine the variations in HMO concentrations from women classified as secretors and nonsecretors, a phenotype that can be identified by the concentration of 2′-fucosylation in their milk. |
format | Online Article Text |
id | pubmed-3983013 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American
Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-39830132015-02-06 Label-Free Absolute Quantitation of Oligosaccharides Using Multiple Reaction Monitoring Hong, Qiuting Ruhaak, L. Renee Totten, Sarah M. Smilowitz, Jennifer T. German, J. Bruce Lebrilla, Carlito B. Anal Chem [Image: see text] An absolute quantitation method for measuring free human milk oligosaccharides (HMOs) in milk samples was developed using multiple reaction monitoring (MRM). To obtain the best sensitivity, the instrument conditions were optimized to reduce the source and postsource fragmentation prior to the quadrupole transmission. Fragmentation spectra of HMOs using collision-induced dissociation were studied to obtain the best characteristic fragments. At least two MRM transitions were used to quantify and identify each structure in the same run. The fragment ions corresponded to the production of singly charged mono-, di-, and trisaccharide fragments. The sensitivity and accuracy of the quantitation using MRM were determined, with the detection limit in the femtomole level and the calibration range spanning over 5 orders of magnitude. Seven commercial HMO standards were used to create calibration curves and were used to determine a universal response for all HMOs. The universal response factor was used to estimate absolute amounts of other structures and the total oligosaccharide content in milk. The quantitation method was applied to 20 human milk samples to determine the variations in HMO concentrations from women classified as secretors and nonsecretors, a phenotype that can be identified by the concentration of 2′-fucosylation in their milk. American Chemical Society 2014-02-06 2014-03-04 /pmc/articles/PMC3983013/ /pubmed/24502421 http://dx.doi.org/10.1021/ac404006z Text en Copyright © 2014 American Chemical Society |
spellingShingle | Hong, Qiuting Ruhaak, L. Renee Totten, Sarah M. Smilowitz, Jennifer T. German, J. Bruce Lebrilla, Carlito B. Label-Free Absolute Quantitation of Oligosaccharides Using Multiple Reaction Monitoring |
title | Label-Free Absolute Quantitation of Oligosaccharides
Using Multiple Reaction Monitoring |
title_full | Label-Free Absolute Quantitation of Oligosaccharides
Using Multiple Reaction Monitoring |
title_fullStr | Label-Free Absolute Quantitation of Oligosaccharides
Using Multiple Reaction Monitoring |
title_full_unstemmed | Label-Free Absolute Quantitation of Oligosaccharides
Using Multiple Reaction Monitoring |
title_short | Label-Free Absolute Quantitation of Oligosaccharides
Using Multiple Reaction Monitoring |
title_sort | label-free absolute quantitation of oligosaccharides
using multiple reaction monitoring |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3983013/ https://www.ncbi.nlm.nih.gov/pubmed/24502421 http://dx.doi.org/10.1021/ac404006z |
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