High-Performance Liquid Chromatography Interfaced with Fluorescence Line-Narrowing Spectroscopy for On-Line Analysis

We have demonstrated, for the first time, that high-performance liquid chromatography (HPLC) can be interfaced with fluorescence line-narrowing spectroscopy (FLNS) for on-line identification and characterization of analytes. Interfacing centered primarily on the design and construction of a novel li...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2001-03, Vol.73 (5), p.951-956
Main Authors: Roberts, Kenneth P, Jankowiak, Ryszard, Small, Gerald J
Format: Article
Language:English
Subjects:
Analytical chemistry
Animals
Benzopyrenes - analysis
Benzopyrenes - toxicity
Carcinogens - analysis
Carcinogens - toxicity
Chemistry
Chromatographic methods and physical methods associated with chromatography
Chromatography
Chromatography, High Pressure Liquid - instrumentation
DNA Adducts - analysis
Exact sciences and technology
Female
Fluorescence
Indicators and Reagents
Mice
Online Systems
Other chromatographic methods
Polycyclic Aromatic Hydrocarbons - analysis
Skin - chemistry
Skin - drug effects
Spectrometry, Fluorescence - methods
Spectrum analysis
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Summary:We have demonstrated, for the first time, that high-performance liquid chromatography (HPLC) can be interfaced with fluorescence line-narrowing spectroscopy (FLNS) for on-line identification and characterization of analytes. Interfacing centered primarily on the design and construction of a novel liquid helium cryostat that accommodates variable-sized quartz tubes/capillaries suitable for HPLC as well as capillary electrophoresis/electrochromatography. In addition to the high spectral resolution afforded by FLNS, analyzing the separated components at 4.2 K minimizes photodegradation from the excitation source and provides indefinite detection times for signal averaging. The proof-of-principle for the HPLC−FLNS system is first demonstrated with a mixture of four structurally similar polycyclic aromatic hydrocarbons and then applied to the analysis of DNA adducts from mouse skin exposed to the carcinogen dibenzo[a,l]pyrene. With femtomole detection limits, HPLC−FLNS can be used for real-world analyses of complex mixtures.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac0008789