Off-line MALDI and ICP MS detection of a single record of
metallothionein separation

a 2013

Off-line MALDI and ICP MS detection of a single record of metallothionein separation

TOMALOVÁ, Iva; Ondřej POLANSKÝ; Pavla FOLTYNOVÁ; Radka MILLIONOVÁ; Jan PREISLER et al.

Základní údaje

Originální název

Off-line MALDI and ICP MS detection of a single record of metallothionein separation

Autoři

TOMALOVÁ, Iva; Ondřej POLANSKÝ; Pavla FOLTYNOVÁ; Radka MILLIONOVÁ a Jan PREISLER

Vydání

European Winter Conference on Plasma Spectrochemistry, 2013

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10406 Analytical chemistry

Stát vydavatele

Polsko

Utajení

není předmětem státního či obchodního tajemství

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/13:00066028

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

mass spectrometry; off-line coupling; separation; metalloproteomics

Změněno: 12. 12. 2013 14:36, Mgr. Iva Benešová, Ph.D.

Anotace

V originále

It is estimated that one third of all proteins require metals to carry out their functions. Therefore, the analyses of native metal-protein and metal-metabolite complexes by hyphenated techniques that combine separation with a specific detection is of growing importance. A new concept of an off-line multidetection platform for metalloprotein/metallopeptide analysis of fractions collected from a single run of capillary electrophoresis (CE) or liquid chromatography (LC) is presented. The separation record, i.e. fractions collected on a target, are analyzed sequentially with MALDI MS and SALD ICP MS. Thus, a single separation can yield both proteomic and metallomic information. Moreover, the off-line nature of the analysis provides further possibilities, such as laser-induced fluorescence detection or on target digestion. Here, the approach is demonstrated on the successful reverse phase HPLC-ESI/MALDI/SALD ICP MS analysis of purified rabbit liver metallothionein (MT) isoforms: The MT1a, MT2d and MT2e were eluted out of a C18 column with ammonium acetate buffer (pH 7.5)/ammonium acetate buffer (pH 7.5) in 60% methanol at 27.8, 29.0 and 33.4 min, respectively. Using a flow splitter, the MT isoforms were detected by on-line ESI–TOF MS and the fractions of the separation were simultaneously collected on a suitable target for successive off-line detection by MALDI and SALD ICP MS. CE was successfully coupled to both MALDI and SALD ICP MS. It can offer enhancement of separation efficiency, modified separation selectivity and high sensitivity. On the other hand, low amount of injected samples poses a challenge for both MALDI and SALD ICP MS detection modes. Initial results of CE as a prospective method for separation of MT isoforms will be presented. The influence of liquid junction interface, fraction collector and MALDI-TOF MS settings and other analysis parameters is discussed in detail.

Návaznosti

ED1.1.00/02.0068, projekt VaV

Název: CEITEC - central european institute of technology

GAP206/12/0538, projekt VaV

Název: Nové strategie úpravy vzorků pro desorpční hmotnostní spektrometrii

Investor: Grantová agentura ČR, Nové strategie úpravy vzorků pro desorpční hmotnostní spektrometrii

GA203/09/1025, projekt VaV

Název: Multidetekční platforma pro mikrokolonové separace proteinů a peptidů

Investor: Grantová agentura ČR, Multidetekční platforma pro mikrokolonové separace proteinů a peptidů

Citovat

TOMALOVÁ, Iva; Ondřej POLANSKÝ; Pavla FOLTYNOVÁ; Radka MILLIONOVÁ a Jan PREISLER. Off-line MALDI and ICP MS detection of a single record of metallothionein separation. In European Winter Conference on Plasma Spectrochemistry. 2013.

@proceedings{1089918,
   author = {Tomalová, Iva and Polanský, Ondřej and Foltynová, Pavla and Millionová, Radka and Preisler, Jan},
   booktitle = {European Winter Conference on Plasma Spectrochemistry},
   keywords = {mass spectrometry; off-line coupling; separation; metalloproteomics},
   language = {eng},
   title = {Off-line MALDI and ICP MS detection of a single record of metallothionein separation},
   year = {2013}
}

TY  - CONF
ID  - 1089918
AU  - Tomalová, Iva - Polanský, Ondřej - Foltynová, Pavla - Millionová, Radka - Preisler, Jan
PY  - 2013
TI  - Off-line MALDI and ICP MS detection of a single record of metallothionein separation
KW  - mass spectrometry
KW  - off-line coupling
KW  - separation
KW  - metalloproteomics
N2  - It is estimated that one third of all proteins require metals to carry out their functions. Therefore, the analyses of native metal-protein and metal-metabolite complexes by hyphenated techniques that combine separation with a specific detection is of growing importance. A new concept of an off-line multidetection platform for metalloprotein/metallopeptide analysis of fractions collected from a single run of capillary electrophoresis (CE) or liquid chromatography (LC) is presented. The separation record, i.e. fractions collected on a target, are analyzed sequentially with MALDI MS and SALD ICP MS. Thus, a single separation can yield both proteomic and metallomic information. Moreover, the off-line nature of the analysis provides further possibilities, such as laser-induced fluorescence detection or on target digestion. Here, the approach is demonstrated on the successful reverse phase HPLC-ESI/MALDI/SALD ICP MS analysis of purified rabbit liver metallothionein (MT) isoforms: The MT1a, MT2d and MT2e were eluted out of a C18 column with ammonium acetate buffer (pH 7.5)/ammonium acetate buffer (pH 7.5) in 60% methanol at 27.8, 29.0 and 33.4 min, respectively. Using a flow splitter, the MT isoforms were detected by on-line ESI–TOF MS and the fractions of the separation were simultaneously collected on a suitable target for successive off-line detection by MALDI and SALD ICP MS. CE was successfully coupled to both MALDI and SALD ICP MS. It can offer enhancement of separation efficiency, modified separation selectivity and high sensitivity. On the other hand, low amount of injected samples poses a challenge for both MALDI and SALD ICP MS detection modes. Initial results of CE as a prospective method for separation of MT isoforms will be presented. The influence of liquid junction interface, fraction collector and MALDI-TOF MS settings and other analysis parameters is discussed in detail.
ER  -

TOMALOVÁ, Iva; Ondřej POLANSKÝ; Pavla FOLTYNOVÁ; Radka MILLIONOVÁ a Jan PREISLER. Off-line MALDI and ICP MS detection of a single record of metallothionein separation. In \textit{European Winter Conference on Plasma Spectrochemistry}. 2013.

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