Bioanalytical applications of fluorescence line-narrowing and non-line-narrowing spectroscopy interfaced with capillary electrophoresis and high-performance liquid chromatography

dc.contributor.advisor Gerald J. Small
dc.contributor.author Roberts, Kenneth
dc.contributor.department Chemistry
dc.date 2018-08-25T00:26:37.000
dc.date.accessioned 2020-07-02T05:42:46Z
dc.date.available 2020-07-02T05:42:46Z
dc.date.copyright Mon Jan 01 00:00:00 UTC 2001
dc.date.issued 2001-01-01
dc.description.abstract <p>Capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC) are now routinely used for analysis of complex biomolecular samples. Difficulties with detection and identification of the targeted analytes, e.g., biomarkers for disease, often arise because of the limited sensitivity and/or selectivity of the detection modality. Identification of an analyte based on its migration time is not definitive because of the possibility of interferences from structurally similar molecules. To circumvent these problems we have developed new methodologies that interface CE and HPLC with fluorescence line-narrowing (FLN) and non-line-narrowing (NLN) spectroscopy for on-line detection and characterization. With these detection methods, one achieves the high sensitivity afforded by laser-induced fluorescence (LIF) (&ap;attomole) and high selectivity provided by FLN spectra, whose rich and sharp (&ap;5 cm-1) vibrational structure serves as fingerprints. Fluorescence line-narrowing is operative at temperatures close to 4 K. Thus, a major challenge was the design of compact cryostats that are compatible with the above separation techniques and allow for rapid cooldown (&ap;1 minute). After cooldown the stationary analyte plugs can be interrogated indefinitely, which leads to significantly lower detection limits. Photodegradation of the analytes is also markedly reduced relative to laser excitation at room temperature. Automated translation of the cryostat/capillary relative to the spatially fixed laser excitation beam allows for sequential analysis of the separated analytes. Lower resolution but still informative analyses can be performed at 77 K.;Results are presented for depurinating polycyclic aromatic hydrocarbons adducted to DNA isolated from the urine of humans exposed to coal smoke, and from the skin of mice treated with the most potent chemical carcinogen, dibenzo[a,l]pyrene. We anticipate that combining FLN and NLN detection with CE and HPLC will be useful in many more bioanalytical applications.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/rtd/451/
dc.identifier.articleid 1450
dc.identifier.contextkey 6073797
dc.identifier.doi https://doi.org/10.31274/rtd-180813-12247
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath rtd/451
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/77152
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/rtd/451/r_3003266.pdf|||Sat Jan 15 00:20:46 UTC 2022
dc.subject.disciplines Analytical Chemistry
dc.subject.disciplines Medical Toxicology
dc.subject.disciplines Physical Chemistry
dc.subject.disciplines Toxicology
dc.subject.keywords Chemistry
dc.subject.keywords Analytical chemistry
dc.title Bioanalytical applications of fluorescence line-narrowing and non-line-narrowing spectroscopy interfaced with capillary electrophoresis and high-performance liquid chromatography
dc.type article
dc.type.genre dissertation
dspace.entity.type Publication
relation.isOrgUnitOfPublication 42864f6e-7a3d-4be3-8b5a-0ae3c3830a11
thesis.degree.level dissertation
thesis.degree.name Doctor of Philosophy
File
Original bundle
Now showing 1 - 1 of 1
Name:
r_3003266.pdf
Size:
3.9 MB
Format:
Adobe Portable Document Format
Description: