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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Second-order advantage achieved with four-way fluorescence excitation-emission-kinetic data processed by parallel factor analysis and trilinear least-squares. Determination of methotrexate and leucovorin in human urine.

Four-way fluorescence data recorded by following the kinetic evolution of excitation-emission fluorescence matrices (EEMs) have been analyzed by parallel factor analysis and trilinear least-squares algorithms. These methodologies exploit the second-order advantage of the studied data, allowing analyte concentrations to be estimated even in the presence of an uncalibrated fluorescent background. They were applied to the simultaneous determination of the components of the anticancer combination of methotrexate and leucovorin in human urine samples. Both analytes were converted into highly fluorescent compounds by oxidation with potassium permanganate, and the kinetics of the reaction was continuously monitored by recording full EEM of the samples at different reaction times. A commercial fast scanning spectrofluorometer has been used for the first time to measure the four-way EEM kinetic data. The rapid scanning instrument allows the acquisition of a complete EEM in 12 s at a wavelength scanning speed of 24 000 nm/min. The emission spectra were recorded from 335 to 490 nm at 5-nm intervals, exciting from 255 to 315 nm at 6-nm intervals. Ten successive EEMs were measured at 72-s intervals, to follow the fluorescence kinetic evolution of the mixture components. Good recoveries were obtained in synthetic binary samples and also in spiked urine samples. The excitation, emission, and kinetic time profiles recovered by both chemometric techniques are in good agreement with experimental observations.[1]


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