Video fluorometer

C.N. Ho, G.D. Christian and E.R. Davidson, Application of the method of rank annihilation to quantitative analysis of multicomponent fluorescence data from the video fluorometer. Anal. Chem., 52 (1980) 1108-1113. 

In thTs communication, we will illustrate the application of the Video Fluorometer to simultaneous analysis of polycyclic aromatic hydrocarbons and warfarin and its metabolite 7-OH warfarin. We hope to convince the reader of the utility of "higher-order" strategies in fluorescence analysis made practicable by imaging detector based instrumentation. 

Video fluorometers digital photometry system, diagram. 

Instrumentation. The Video Fluorometer described above was used to acquire all fluorescence data. The excitation and emission slits were selected to give a band pass of 6.6 nm. All data was acquired at a resolution of 64 X 64 pixels and is the result of summation of 512 frames to memory followed by subtraction of 512 dark frames. The data acquisition, display and analysis programs were written in PDP-11 Fortran IV under RT-11 and the listings are contained in the first author s Ph.D. dissertation (1 ). 

Figure 4. Data acquisition with the Video Fluorometer. (A) Single frame EEM for 1 X 10" M perylene using 64 X 64 format (B) EEM for same sample after summation of 512 frames to memory (C) As in B hut with subtraction of 512 dark frames (D) As in C but with subtraction of solvent blank.

Using the Video Fluorometer, we obtained a series of standard intensity values for the individual standard compounds (warfarin, 7-OH warfarin) at known concentrations under acidic and basic conditions. Then a series of mixtures of known composition was made up, and intensity values were obtained for these under acidic and basic conditions. The concentrations of the individual components of the mixtures was then determined by Equation 3. 

Super-resolution in chromatography is defined as the ability to detect the presence of two or more components when their mean elution tines fall within two standeurd peak widths of each other. Numerical deconvolution techniques for achieving superresolution eu e considered for the case of a single channel detector (7ZD), a multichannel detector (HS) and a two dimensional detector (video fluorometer). It is shown that the degree of ambiguity of the deconvolution decreases in going from a monochannel detector to a multicdiannel detector to a multidimensional detector. Three types of analytical situations are considered for each type of detector ... 

Ho CN, Christian GD, Davidson ER, Simultaneous multicomponent rank annihilation and applications to multicomponent fluorescent data acquired by the video fluorometer, Analytical Chemistry, 1981, 53, 92-98. 

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