Improved analyte detectability of proteins and peptide lysates by means of multiple large-volume injection in LC-MS

A 'multiple (trapping) large-volume injection' approach was developed for the analysis of peptides and proteins. In this way, a maximally 10-fold gain in sensitivity could be achieved. The system involves the use of an automated 10-port switching valve in combination with a 1 mm i.d. trapp...

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Bibliographic Details
Published in:	Journal of separation science Vol. 32; no. 14; pp. 2346 - 2352
Main Authors:	Storme, Michael L, t'Kindt, Ruben S, Van Bocxlaer, Jan F
Format:	Journal Article
Language:	English
Published:	Weinheim Wiley-VCH Verlag 01-07-2009 WILEY-VCH Verlag WILEY‐VCH Verlag Wiley
Subjects:	(Absolute) Protein quantitation Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Calibration Cattle Chickens Chromatography, High Pressure Liquid - methods Cystatin C - analysis Cystatins - analysis Fundamental and applied biological sciences. Psychology General aspects, investigation methods Hemoglobins - analysis Large-volume injection LC-MS(/MS) Linear Models Mass Spectrometry - methods Peptides Peptides - analysis Proteins Sensitivity Sensitivity and Specificity Tandem mass spectrometry Chemical analysis Peptides Coupled method Multiple reaction monitoring mode HPLC chromatography Detectability (Absolute) Protein quantitation Electrospray Protein Large volume injection Sensitivity Analysis method Peptide fragment LC-MS(/MS) Large-volume injection Detection Mass spectrometry Quantitative analysis
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Summary:	A 'multiple (trapping) large-volume injection' approach was developed for the analysis of peptides and proteins. In this way, a maximally 10-fold gain in sensitivity could be achieved. The system involves the use of an automated 10-port switching valve in combination with a 1 mm i.d. trapping/guard column and a 1 mm i.d.x150 mm analytical column. The optimized multiple injection/loading procedure allows quantitative measurements of peptides and protein lysates. Linear calibration curves (R² [greater, not equals] 0.988) over a minimum of two orders of magnitude were generated for a range of peptide and protein standards with sensitivities equal to or even exceeding, those generally achieved only through increasing miniaturization (quantification limit [greater, not equals]0.5 pmol/mL).
Bibliography:	http://dx.doi.org/10.1002/jssc.200900131 Bijzonder OnderzoeksFonds of the Ghent University - No. B/06859-BOF06/24j/025 istex:3CDB8183D341DA61B3ED605E0D4200BBE56E8DDE FWO-Vlaanderen - No. G.0320.0 ark:/67375/WNG-GP72VDNM-3 ArticleID:JSSC200900131 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	1615-9306 1615-9314
DOI:	10.1002/jssc.200900131