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

• 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
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac0008789

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.