| Advantages and limitations of coupling isotachophoresis and comprehensive isotachophoresis-capillary electrophoresis to time-of-flight mass spectrometry. | |
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MedLine Citation:
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PMID: 12735473 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Capillary isotachophoresis (ITP) and comprehensive isotachophoresis-capillary electrophoresis (ITP-CE) were successfully coupled to electrospray ionization (ESI) orthogonal acceleration time-of-flight mass spectrometry (TOF-MS) using angiotensin peptides as model analytes. The utility of ITP-TOF-MS and ITP-CE-TOF-MS for the analysis of samples containing analyte amounts sufficient to form flat-top ITP zones (30 microM) as well as for samples with trace analyte amounts (0.3 microM) was studied. Separations were performed in 150 microm internal diameter (I.D.) capillaries for the ITP experiments, and in 200 microm I.D. (ITP) and 50 microm I.D. (CE) capillaries for ITP-CE experiments. The fused-silica columns were coated with poly(vinyl alcohol) to suppress electroosmotic flow that can disrupt ITP zone profiles. The sample loading capacity in both ITP and comprehensive ITP-CE was greatly enhanced (up to 10 microl) compared with typical nanoliter-sized injection volumes in CE. It was concluded that ITP-TOF-MS alone was adequate for the separation and detection of high concentration samples. The outcome was different at lower analyte concentrations where mixed zones or very sharp peaks formed. With formation of mixed zones, ion suppression and discrimination could occur, complicating quantitative determination of the analytes. This problem was effectively overcome by inserting a CE capillary between the ITP and TOF-MS. In such an arrangement, samples were preconcentrated in the high load WTP capillary and then injected into a CE capillary where they were separated into non-overlapping peaks prior to their detection by TOF-MS. The advantage of this comprehensive arrangement, which we have described previously, is that there is no need to discard portions of the sample in order to avoid overloading of the CE capillary. The whole sample is analyzed by multiple injections from ITP to CE. Thus, this method can be used for the analysis of complex samples with wide ranges of component concentrations. |
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Authors:
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Zlatuse D Peterson; Christopher R Bowerbank; David C Collins; Steven W Graves; Milton L Lee |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Journal of chromatography. A Volume: 992 ISSN: 0021-9673 ISO Abbreviation: J Chromatogr A Publication Date: 2003 Apr |
Date Detail:
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Created Date: 2003-05-08 Completed Date: 2004-01-28 Revised Date: 2009-01-15 |
Medline Journal Info:
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Nlm Unique ID: 9318488 Medline TA: J Chromatogr A Country: Netherlands |
Other Details:
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Languages: eng Pagination: 169-79 Citation Subset: IM |
Affiliation:
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Department of Chemistry and Biochemistry, Brigham Young University, C267 Benson Science Building, PO Box 25700, Provo, UT 84602-5700, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Electrophoresis
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methods* Electrophoresis, Capillary / methods* Spectrometry, Mass, Electrospray Ionization / methods* |
| Grant Support | |
ID/Acronym/Agency:
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1R43RR14936-01/RR/NCRR NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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