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Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography-electrospray ionisation-mass spectrometry

Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography-electrospray ionisation-mass spectrometry
Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography-electrospray ionisation-mass spectrometry
Low molecular weight (LMW; < 1 kDa) dissolved organic nitrogen (DON) is the dominant DON fraction in marine systems. However, existing methods for DON characterization exclude this fraction through size exclusion or destruction during sample hydrolysis. In this study, we developed a method for the extraction and analysis of LMW peptides in saline waters. The procedure involves a solid phase extraction (SPE) preconcentration step using a polystyrene-divinylbenzene sorbent, then elution and detection by liquid chromatographyelectrospray ionization-mass spectrometry (LC/ESI/MS). The procedure was tested with 1-L water samples containing peptides (10 ng L–1) ranging from 188.2-1946.0 Da. The analytes were characterized using “top-down” sequencing to confirm their structure, and the method was then successfully applied to saline water samples extracted from a mono-algal culture of Phaeodactylum tricornutum. This enabled a number of unknown analytes to be partially sequenced (complete sequencing should be possible using additional MSn scans and larger sample volumes). The method allows for the detection of peptides at the ng L–1 level and further preconcentration is possible. The SPE step allowed determination of peptides from saline water, a matrix incompatible with direct LC/MS analysis. The method can, therefore, be used in DON characterization studies.
1541-5856
52-63
Curtis-Jackson, Philippa K.
74fd2eb2-7434-4560-a2c5-8a4402bd5e79
Massé, Guillaume
1453cc47-02d2-4831-bab1-f743a861a587
Gledhill, Martha
da795c1e-1489-4d40-9df1-fc6bde54382d
Fitzsimons, Mark F.
52a13739-d138-4274-b9e5-c8494cbfa374
Curtis-Jackson, Philippa K.
74fd2eb2-7434-4560-a2c5-8a4402bd5e79
Massé, Guillaume
1453cc47-02d2-4831-bab1-f743a861a587
Gledhill, Martha
da795c1e-1489-4d40-9df1-fc6bde54382d
Fitzsimons, Mark F.
52a13739-d138-4274-b9e5-c8494cbfa374

Curtis-Jackson, Philippa K., Massé, Guillaume, Gledhill, Martha and Fitzsimons, Mark F. (2009) Characterization of low molecular weight dissolved organic nitrogen by liquid chromatography-electrospray ionisation-mass spectrometry. Limnology and Oceanography: Methods, 7, 52-63.

Record type: Article

Abstract

Low molecular weight (LMW; < 1 kDa) dissolved organic nitrogen (DON) is the dominant DON fraction in marine systems. However, existing methods for DON characterization exclude this fraction through size exclusion or destruction during sample hydrolysis. In this study, we developed a method for the extraction and analysis of LMW peptides in saline waters. The procedure involves a solid phase extraction (SPE) preconcentration step using a polystyrene-divinylbenzene sorbent, then elution and detection by liquid chromatographyelectrospray ionization-mass spectrometry (LC/ESI/MS). The procedure was tested with 1-L water samples containing peptides (10 ng L–1) ranging from 188.2-1946.0 Da. The analytes were characterized using “top-down” sequencing to confirm their structure, and the method was then successfully applied to saline water samples extracted from a mono-algal culture of Phaeodactylum tricornutum. This enabled a number of unknown analytes to be partially sequenced (complete sequencing should be possible using additional MSn scans and larger sample volumes). The method allows for the detection of peptides at the ng L–1 level and further preconcentration is possible. The SPE step allowed determination of peptides from saline water, a matrix incompatible with direct LC/MS analysis. The method can, therefore, be used in DON characterization studies.

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Published date: 2009

Identifiers

Local EPrints ID: 65600
URI: http://eprints.soton.ac.uk/id/eprint/65600
ISSN: 1541-5856
PURE UUID: 9ccebcfa-c741-4f5a-a64d-8a01785165ea

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Date deposited: 25 Feb 2009
Last modified: 11 Sep 2020 16:31

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