Limitations of synchronous luminescence spectrometry in multicomponent analysis

Latz, Howard W.; Ullman, Alan H.; Winefordner, J. D.

doi:10.1021/ac50036a050

Cited by 28 publications

(10 citation statements)

References 6 publications

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“…A fluorescence signal at 374 nm exists only 5 nm from the signature signal (379 nm) at shorter wavelengths, but it is impossible without further experimentation to determine the identity of this signal ( 374 , 34 nm). Even if it were possible to assign the origin of this signal to BP-7,10/8,9-tetrol, its position in the spectrum and relative magnitude would probably vary as a function of the composition of the complex mixture under study (32).…”

Section: Resultsmentioning

confidence: 99%

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Derivative fluorescence spectral analysis of polycyclic aromatic hydrocarbon-DNA adducts in human placenta

Weston¹,

Manchester²,

Poirier³

et al. 1989

Chem. Res. Toxicol.

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Metabolic activation in humans of chemical carcinogens found in the environment results in the formation of carcinogen-DNA adducts in vivo. Some polycyclic aromatic hydrocarbon-DNA adducts in human DNA can be hydrolyzed under mildly acidic conditions to yield tetrahydrotetrol derivatives which may then be detected by synchronous fluorescence spectroscopy. In an analysis of human placental DNA, second derivative spectroscopy alone was unable to resolve the synchronous fluorescent signature for r-7,t-8,t-9,c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[alpha]pyrene from a crude extract, because a complex array of other fluorescent materials was also present. Purification of the sample by a combination of chromatographic procedures including immunoaffinity chromatography and HPLC has now been shown to yield r-7,t-8,t-9,c-10-tetrahydroxy-7,8,9,10-tetrahydrobenzo[alpha]pyrene residues from human DNA that are spectroscopically pure at the second derivative level. Immunoaffinity columns were prepared with rabbit antiserum raised against DNA that had been modified with (+/-)-r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[alpha]pyre ne. This antiserum has now been shown to recognize DNA samples that have been modified with six different polycyclic aromatic hydrocarbon diol epoxides and is probably only specific for a broad spectrum of polycyclic aromatic hydrocarbon-DNA adducts. Adducts were eluted from the immunoaffinity columns, hydrolyzed with acid, and extracted into isoamyl alcohol, before being subjected to high-performance liquid chromatography. These experiments reveal important limitations of second derivative fluorescence spectroscopy as a tool in the analysis of complex environmental mixtures. Furthermore, they extensively define the ability of anti-benzo[alpha]pyrenediol epoxide-DNA antibodies to recognize different types of polycyclic aromatic hydrocarbon-DNA adducts.

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Section: Resultsmentioning

confidence: 99%

“…These data support previous HPLC/SFS analyses (20) of DNA isolated from the peripheral blood lymphocytes of coke oven workers where only zeroth-order SFS were considered. However, the presence of other large fluorescent signals at longer wavelengths may have implications for quantitative analysis (32).…”

Section: Resultsmentioning

confidence: 99%

Derivative fluorescence spectral analysis of polycyclic aromatic hydrocarbon-DNA adducts in human placenta

Weston¹,

Manchester²,

Poirier³

et al. 1989

Chem. Res. Toxicol.

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“…The SyF spectrum may sample only a section of a uorescence peak or m ight even miss a uorophore completely, depending on what constant offset (Dl) value is used. 25 Therefore, because Dl is not likely to be optimum for all peaks in any given experiment, it rem ains uncertain if the SyF spectra of FA are sensitive enough to measure the total possible quenching response due to copper addition. For these reasons, a m ulti-wavelength m odel (M WM ) is developed to use the uorescence response at m ultiple wavelengths to quantify copper binding by FA.…”

Section: Introductionmentioning

confidence: 99%

Multi-Wavelength Fluorescence-Quenching Model for Determination of Cu²⁺ Conditional Stability Constants and Ligand Concentrations of Fulvic Acid

Hays

Ryan²,

Pennell³

2003

Appl Spectrosc

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In this work, a multi-wavelength model (MWM) is developed. It uses fluorescence bands in the fulvic acid (FA) spectrum that quench upon binding of inorganic Cu2+ to FA. Quenching data at pH values of 5, 6, and 7 are placed in sets, containing fluorescence measures at select wavelengths versus added copper (CM). Intensity data of wavelength set 1 are obtained from 25 nm constant offset synchronous fluorescence spectra (SyF), in which are observed distinct peaks (lambda(ex) = 415 nm, lambda(em) = 440 nm; and lambda(ex) = 471 nm, lambda(em) = 496 nm). Wavelength set 2 intensity data are obtained from the FA fluorescence excitation and emission maxima (lambda(ex) = 335 nm, lambda(em) = 450 nm; and X(ex) = 471 nm, lambda(em) = 496 nm). Application of MWM shows that the multi-wavelength data sets characterize ligands of different binding strength (log K(x)) and concentration (C(Lx)). Corresponding to pH values of 5, 6, and 7, mean and standard deviation values for wavelength set 1 are log K(415/440) = 4.66 (0.12), 5.03 (0.12), and 5.05 (0.08), log K(471/496) = 4.93 (0.06), 5.27 (0.11), and 5.39 (0.09), C(415/440) = 3.1 (1.5), 10.9 (4.5), and 7.9 (3.9) microM, C(471/496) = 14.3 (3.0), 1.7 (0.6), and 1.4 (0.5) microM. And for wavelength set 2, log K(335/450) = 4.50 (0.03), 4.96 (0.27), and 5.22 (0.08), log K(471/496) = 5.02 (0.04), 5.42 (0.32), and 5.71 (0.09), C(335/450) = 8.8 (0.5), 21.9 (7.9), and 18.7 (0.3) microM, C(471/496) = 21.0 (2.5), 7.17 (1.2), and 7.09 (0.3) micrpM. The ability of the 415/440 nm SyF transect to characterize the main excitation and emission maximum of FA at 335/440 nm is evaluated. Relatively low concentration values returned by the model for this transect (415/440 nm) suggest that it is not entirely illustrative of the maximum. The model predictive capability is verified at pH 6 with two fluorescing Cu2+ chelating organic compounds, L-tyrosine and salicylic acid. This test confirms that the model is capable of providing good estimates of equilibrium binding parameters from multi-wavelength measurements of a mixed ligand system.

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“…This and the still broad bands result in many interferences in any complex mixture (26 (27)(28)(29). In other types of matrices, these same vibronic transitions can be considerably sharper, vide infra (31).…”

Section: Synchronously Scanned Luminescencementioning

confidence: 99%

Fluorescence line narrowing spectrometry of polycyclic aromatic hydrocarbons in organic glasses

Brown

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Limitations of synchronous luminescence spectrometry in multicomponent analysis

Cited by 28 publications

References 6 publications

Derivative fluorescence spectral analysis of polycyclic aromatic hydrocarbon-DNA adducts in human placenta

Derivative fluorescence spectral analysis of polycyclic aromatic hydrocarbon-DNA adducts in human placenta

Multi-Wavelength Fluorescence-Quenching Model for Determination of Cu²⁺ Conditional Stability Constants and Ligand Concentrations of Fulvic Acid

Fluorescence line narrowing spectrometry of polycyclic aromatic hydrocarbons in organic glasses

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Limitations of synchronous luminescence spectrometry in multicomponent analysis

Cited by 28 publications

References 6 publications

Derivative fluorescence spectral analysis of polycyclic aromatic hydrocarbon-DNA adducts in human placenta

Derivative fluorescence spectral analysis of polycyclic aromatic hydrocarbon-DNA adducts in human placenta

Multi-Wavelength Fluorescence-Quenching Model for Determination of Cu2+ Conditional Stability Constants and Ligand Concentrations of Fulvic Acid

Fluorescence line narrowing spectrometry of polycyclic aromatic hydrocarbons in organic glasses

Contact Info

Product

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Multi-Wavelength Fluorescence-Quenching Model for Determination of Cu²⁺ Conditional Stability Constants and Ligand Concentrations of Fulvic Acid