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 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.
Roberts
K P
KP
Ames Laboratory--USDOE, Iowa 50011, USA.
Jankowiak
R
R
Small
G J
GJ
D000818:Animals; D001580:Benzopyrenes / Q000032:analysis / Q000633:toxicity; D002273:Carcinogens / Q000032:analysis / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D018736:DNA Adducts / Q000032:analysis; D005260:Female; D007202:Indicators and Reagents; D051379:Mice; D009862:Online Systems; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis; D012867:Skin / Q000737:chemistry / Q000187:drug effects; D013050:Spectrometry, Fluorescence / Q000379:methods*
D001580:Benzopyrenes; D002273:Carcinogens; D018736:DNA Adducts; D007202:Indicators and Reagents; D011084:Polycyclic Aromatic Hydrocarbons; C041517:dibenzo(a,l)pyrene