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

被引：12

作者：

Curtis-Jackson, Philippa K. ^{[1
]}

Masse, Guillaume ^{[1
,2
]}

Gledhill, Martha ^{[3
]}

Fitzsimons, Mark F. ^{[1
]}

机构：

[1] Univ Plymouth, Sch Earth Ocean & Environm Sci, Petr & Environm Geochem Grp, Plymouth PL4 8AA, Devon, England

[2] LOCEAN IPSL, UMR 7159, F-75252 Paris, France

[3] Univ Southampton, Natl Oceanog Ctr, Sch Ocean & Earth Sci, Southampton SO14 3ZH, Hants, England

来源：

LIMNOLOGY AND OCEANOGRAPHY-METHODS | 2009年 / 7卷

基金：

英国自然环境研究理事会;

关键词：

UNDERIVATIZED SMALL PEPTIDES; AMINO-ACIDS; CHEMICAL-CHARACTERIZATION; FULVIC-ACIDS; PROTONATED PEPTIDES; ESTUARINE WATERS; PROTEINS; MATTER; SEDIMENTS; BIOAVAILABILITY;

D O I：

10.4319/lom.2009.7.52

中图分类号：

Q [生物科学];

学科分类号：

090105 [作物生产系统与生态工程];

摘要：

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 chromatography-electrospray 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 MS(n) 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.

引用

页码：52 / 63

页数：12

共 49 条

[1]

Separation of underivatized small peptides on a porous graphitic carbon column by ion-pair chromatography and evaporative light scattering detection [J].

Adoubel, AA ;

Guenu, S ;

Elfakir, C ;

Dreux, M .

JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES, 2000, 23 (16) :2433-2446

[2]

Distributions and seasonal variability of dissolved organic nitrogen in two estuaries in SW England [J].

Badr, El-Sayed A. ;

Tappin, Alan D. ;

Achterberg, Eric P. .

MARINE CHEMISTRY, 2008, 110 (3-4) :153-164

[3]

Abundance, size distribution, and stable carbon and nitrogen isotopic compositions of marine organic matter isolated by tangential-flow ultrafiltration [J].

Benner, R ;

Biddanda, B ;

Black, B ;

McCarthy, M .

MARINE CHEMISTRY, 1997, 57 (3-4) :243-263

[4]

Dissolved organic nitrogen hydrolysis rates in axenic cultures of Aureococcus anophagefferens (Pelagophyceae):: Comparison with heterotrophic bacteria [J].

Berg, GM ;

Repeta, DJ ;

Laroche, J .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (01) :401-404

[5]

Bier M.E., 1997, EIECTROSPRAY IONISAT, P235

[6]

DON as a source of bioavailable nitrogen for phytoplankton [J].

Bronk, D. A. ;

See, J. H. ;

Bradley, P. ;

Killberg, L. .

BIOGEOSCIENCES, 2007, 4 (03) :283-296

[7]

Bronk D.A., 2002, Biogeochemistry of marine dissolved organic matter, P153, DOI [DOI 10.1016/B978-012323841-2/50007-5, 10.1016/b978-012323841-2/50007-5]

[8]

De novo peptide sequencing via tandem mass spectrometry [J].