| Analysis of Oligonucleotides by HPLC-Electrospray Ionization Mass Spectrometry. | |
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MedLine Citation:
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PMID: 21639339 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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A new interface procedure has been developed that allows, for the first time, the high-efficiency analysis of synthetic oligonucleotides up to 75 bases by reversed-phase HPLC and on-line electrospray ionization mass spectrometry. For oligonucleotides up to 30 bases in length, single-base resolution can be obtained with low levels of cation adduct formation in the negative ion electrospray mass spectra. A key part of the method uses 1,1,1,3,3,3-hexafluoro-2-propanol as an additive to the HPLC mobile phase, adjusted to pH 7.0 with triethylamine. This novel additive results in both good HPLC separation and efficient electrospray ionization. The broad potential of this new method is demonstrated for synthetic homopolymers of thymidine (PolyT), fragments based on the pBR322 plasmid sequence, and phosphorothioate ester antisense oligonucleotides. This approach will be of particular utility for the characterization of DNA probes and PCR primers and quality control of antisense compounds such as phosphorothioates and their metabolites, as well as of materials used in clinical trials. |
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Authors:
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A Apffel; J A Chakel; S Fischer; K Lichtenwalter; W S Hancock |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Analytical chemistry Volume: 69 ISSN: 0003-2700 ISO Abbreviation: Anal. Chem. Publication Date: 1997 Apr |
Date Detail:
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Created Date: 2011-06-06 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 1320-5 Citation Subset: - |
Affiliation:
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Biomeasurements Group, Hewlett-Packard Laboratories, Palo Alto, California 94304, and California Analytical Division, Hewlett-Packard Company, Palo Alto, California 94304. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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