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Liquid Chromatography – tandem Mass Spectrometry...tandem Mass Spectrometry An Introduction for...
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Liquid Chromatography – tandem Mass Spectrometry An Introduction for Beginners Steven Pauwels Jessa Hospital, 7 th of May 2013
Transcript of Liquid Chromatography – tandem Mass Spectrometry...tandem Mass Spectrometry An Introduction for...
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Liquid Chromatography – tandem Mass Spectrometry
An Introduction for Beginners
Steven Pauwels Jessa Hospital, 7th of May 2013
http://intranet/
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Why show interest?
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Tacrolimus Proficiency testing
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1999 2010 2013
% LC MS
www.bioanalytics.co.uk
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www.pubmed.org
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2004 2012
6772 analyses
Immunosuppresives
11927 analyses
Immunosuppresives, Antiepileptic,
Antifungal, Endocrine
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Steroids Antibiotics?
Metabolites? Cancer therapy?
~ 0,1% van analyses
Evolution UZ Leuven
PresentatorPresentatienotitiesBegin immunosuppresiva: vaak voor TDM/tox: nog geen immunoassay, specificiteit
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Grebe SK, Singh RJ. Clin Biochem Rev 2011
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van den Ouweland JMW, Kema IP. J Chromatography B 2012
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Liquid Chromatography in a Nutshell
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Mobile phase: -RP: polar -NP: apolar
Stationary phase: -RP: apolar -NP: polar
www.wikipedia.com
PresentatorPresentatienotitieshttp://www.hitachi-hitec.com/global/science/lc/lc_basic_1.html
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www.restek.com
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www.phenomenex.com
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www.wikipedia.com
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www.restek.com
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www.restek.com
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Demystifying Mass Spectrometry
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www.wikipedia.com
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www.wikipedia.com
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q F
F
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www.wikipedia.com
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www.wikipedia.com
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www.shimadzu.com
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www.shimadzu.com
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www.shimadzu.com
PresentatorPresentatienotitiesShimadzu
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www.planetorbitrap.com
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Raida M et al. J Am Soc Mass Spectrom 1999
PresentatorPresentatienotitiesLiquid chromatography and electrospray mass spectrometric mapping of peptides from human plasma filtrateManfred Raidaa, , , Peter Schulz-Knappea, Gabriele Heinea, Wolf-Georg Forssmannaa Lower Saxony Institute for Peptide Research, Hannover, GermanyReceived 10 December 1997 Revised 21 July 1998 Accepted 22 August 1998 Available online 15 July 1999 http://dx.doi.org/10.1016/S1044-0305(98)00117-2, How to Cite or Link Using DOIPermissions & Reprints
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Q IT TOF FT OT FT ICR
Resolution - +/- + ++ +++
Mass Accuracy (ppm)
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Acquisition Speed (Hz)
+ + +++ +/- -
m/z range - + +++ + ++
Linearity ++ + + +/- +/-
Price +++ ++ +/- +/- -
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Domon B, Aebersold R. Science 2006
PresentatorPresentatienotitiesMass Spectrometry and Protein AnalysisScience 14 April 2006: �vol. 312 no. 5771 212-217
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Holcapek et al. J Chromatography B 2012
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Ionization: the Missing Link
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Grebe SK, Singh RJ. Clin Biochem Rev 2011
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Grebe SK, Singh RJ. Clin Biochem Rev 2011
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Vogeser M, Seger C. Clin Chem 2010
PresentatorPresentatienotitieswww.molecularstation.com
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Holcapek et al. J Chromatography B 2012
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Practical Example
• Develop a LC-MS/MS method for X
• How to start?
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Sample Preparation
• Dilute and shoot • Protein precipitation • SPE • Liquid-Liquid • IAC
Purity Cost Time
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After Google
• Tuning • Optimalisation/Amelioration • Validation (extended)
– also ME – IS – Linearity – LOQ – Interferences
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www.bioanalytics.com
Liquid Chromatography – tandem Mass SpectrometryWhy show interest?Tacrolimus Proficiency testingDianummer 4Evolution UZ LeuvenDianummer 6Dianummer 7Liquid Chromatography in a NutshellDianummer 9Dianummer 10Dianummer 11Dianummer 12Dianummer 13Dianummer 14Dianummer 15Demystifying Mass SpectrometryDianummer 17Dianummer 18Dianummer 19Dianummer 20Dianummer 21Dianummer 22Dianummer 23Dianummer 24Dianummer 25Dianummer 26Dianummer 27Dianummer 28Ionization: the Missing LinkDianummer 30Dianummer 31Dianummer 32Dianummer 33Practical ExampleDianummer 35Dianummer 36Sample PreparationDianummer 38Dianummer 39After GoogleDianummer 41
