Determination of Cerebrospinal Fluid Proteome Variations by Isobaric Labeling Coupled with Strong Cation-Exchange Chromatography and Tandem Mass Spectrometry

2019 ◽  
pp. 155-168 ◽  
Author(s):  
Mercedes Lachén-Montes ◽  
Andrea González-Morales ◽  
Joaquín Fernández-Irigoyen ◽  
Enrique Santamaría
Keyword(s):  
Mass Spectrometry ◽  
Cerebrospinal Fluid ◽  
Tandem Mass Spectrometry ◽  
Cation Exchange ◽  
Exchange Chromatography ◽  
Tandem Mass ◽  
Cation Exchange Chromatography ◽  
Strong Cation Exchange ◽  
Strong Cation Exchange Chromatography

Analysis of Amadori Compounds by High-Performance Cation Exchange Chromatography Coupled to Tandem Mass Spectrometry

2005 ◽  
Vol 77 (1) ◽  
pp. 140-147 ◽  
Author(s):  
Tomas Davidek ◽  
Karin Kraehenbuehl ◽  
Stéphanie Devaud ◽  
Fabien Robert ◽  
Imre Blank
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Cation Exchange ◽  
High Performance ◽  
Exchange Chromatography ◽  
Tandem Mass ◽  
Cation Exchange Chromatography ◽  
Amadori Compounds

scholarly journals The use of LC tandem mass spectrometry as part of a workflow for the screening and identification of hemoglobin variants. Characterization of Hb Ullevaal as an example

2017 ◽  
Vol 41 (3) ◽  
Author(s):  
Matthias Weber ◽  
Julia J.M. Eekels
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Cation Exchange ◽  
Peripheral Blood ◽  
Globin Gene ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
South Florida ◽  
Tandem Mass ◽  
Genetic Sequencing

AbstractBackground:About 2/3 of the hemoglobin (Hb) variants do not show a charge difference to the wildtype entity but most of them differ in hydrophobicity. In addition to cation exchange chromatography, globin differentiation by liquid chromatography-tandem mass spectrometry (MS) was introduced. Hb Ullevaal was chosen as one example to demonstrate the performance of the approach.Methods:Screening for Hb variants was performed using cation exchange HPLC. For globin separation reversed phase-LC/MS was performed. Tryptic digests of variants were separated on RP-HPLC with or without CID-fragmentation and database search for identification of mutation bearing fragments. Sequencing of the β-globin gene has been performed.Results:HbS, HbC, HbE, Hb South Florida and Hb Ullevaal show typical and distinct patterns in the globin LC/MS according to the theoretical protein data. The tryptic digest of Hb Ullevaal resulted in the identification of the respective mutated peptide βT9, which was confirmed by genetic sequencing.Conclusions:By the application of globin-LC/MS two more dimensions for the Hb identification are added, hydropathicity and protein mass. With this workflow as screening procedure for Hb variants it is expected to be able to detect and identify the majority of variants with the exception of highly unstable variants, which cannot be determined in the peripheral blood at all. A negative result makes the presence of a significant Hb variant in the peripheral blood improbable.

Strong cation-exchange chromatography of proteins on a sulfoalkylated monolithic cryogel

2015 ◽  
Vol 1386 ◽  
pp. 13-21 ◽  
Author(s):  
Işık Perçin ◽  
Rushd Khalaf ◽  
Bastian Brand ◽  
Massimo Morbidelli ◽  
Orhan Gezici
Keyword(s):  
Cation Exchange ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Strong Cation Exchange ◽  
Strong Cation Exchange Chromatography

scholarly journals Determination of S-Adenosylmethionine and S-Adenosylhomocysteine in Plasma and Cerebrospinal Fluid by Stable-Isotope Dilution Tandem Mass Spectrometry

2000 ◽  
Vol 46 (10) ◽  
pp. 1650-1656 ◽  
Author(s):  
Eduard A Struys ◽  
Erwin E W Jansen ◽  
Kees de Meer ◽  
Cornelis Jakobs
Keyword(s):  
Mass Spectrometry ◽  
Cerebrospinal Fluid ◽  
Stable Isotope ◽  
Tandem Mass Spectrometry ◽  
Solid Phase ◽  
Extraction Procedure ◽  
Internal Standard ◽  
Tandem Mass ◽  
Analytical Technique

Abstract Background: Available methods for the determination of nanomolar concentrations of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) in plasma and cerebrospinal fluid (CSF) are time-consuming. We wished to develop a method for their rapid and simultaneous measurement. Methods: We used tandem mass spectrometry (MS/MS) for the simultaneous determination of SAM and SAH, with stable-isotope-labeled internal standards. The 13C5-SAH internal standard was enzymatically prepared using SAH-hydrolase and [13C5]adenosine. The method comprises a weak anion-exchange solid-phase extraction procedure serving as clean-up step for the deproteinized plasma and CSF samples. After clean-up, samples were injected on a C18 HPLC column, which was connected directly to the tandem mass spectrometer, operating in MS/MS mode. Results: In plasma samples, the intraassay CVs for SAM and SAH were 4.2% and 4.0%, respectively, and the interassay CVs were 7.6% and 5.9%, respectively. In CSF, the intraassay CVs for SAM and SAH were 6.8% and 6.9%, respectively, and the interassay CVs were 4.2% and 5.5%, respectively. Mean recovery of SAM and SAH for both matrices at two concentrations was 93%. Detection limits for SAM and SAH in samples were 7.5 and 2.5 nmol/L, respectively. Concentrations of SAM and SAH in plasma from healthy subjects were within the previously reported ranges. In 10 CSF samples, the mean concentrations (range) were 248 (137–385) nmol/L for SAM and 11.3 (8.9–14.1) nmol/L for SAH. Conclusions: SAM and SAH can be analyzed by MS/MS, taking optimal advantage of the speed and high sensitivity and specificity of this relatively new analytical technique.

Sign in / Sign up

Export Citation Format

Share Document