Author： Victor Mark A. Crouch Stanley R.
Publisher： Society for Applied Spectroscopy
ISSN： 0003-7028
Source： Applied Spectroscopy, Vol.49, Iss.7, 1995-07, pp. : 1041-1047
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Abstract
Synchronous fluorescence spectroscopy is frequently used to resolve multiple components in mixtures without separation. However, uncompensated inner-filter effects can lead to unexpected and/or large quantitative errors. An instrument and a method that correct for primary and secondary inner-filter effects in synchronous fluorescence spectroscopy have been designed and characterized. A bifurcated fiber-optic-based diode array fluorometer that simultaneously measures front-surface fluorescence and absorbance was constructed. A transfer function that mathematically describes the coupling efficiency between the excitation and emission legs of the fiber optics is defined and characterized. Once characterized, such a fluorometer is capable of calculating what the fluorescence measurement would be in the absence of inner-filter effects. Several experimental procedures for determining the instrument's transfer function are compared and discussed. Applications of the various procedures to experimental fluorescence inner-filter corrections to problematic samples are demonstrated.
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