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EXPERIMENTAL PROCEDURE 1. Verify the aborbance baseline by recording the absorbance spectrum between 350 and 410 nm with phosphate buffer in both the analytical and reference cuvettes. 2. JlOO). Replace the cuvette in the analytical compartment of the spectrophotome- 28 ter and record the absorbance spectrum using a vertical scale that allows easy reading of the absorbance values in the range 350-410 nm. Determine the absorbance peak wavelength accurately and call it \. 3. • 4. 5. Empty the measuring cuvette by pouring the contents into the beaker containing the 111M solution.

Time- and frequency-domain analysis based on fast detection and averaging of the detector output or on single-photon timing will be reviewed. Criteria for choosing among commercially available equipment, as well as for implementing home-made systems, will be examined. , Basic Photophysics The first step in the activation process of a photochemical reagent or a photosensitizer is the photoexcitation of the reagent to its lowest excited singlet state, S I' From there, it can be transformed by a chemical reaction, undergo intersystem crossing to the triplet state, T1' or decay back to the ground'state, So' via nonradiative decay pathways or fluorescence emission.

The scattered excitation pulse is measured and averaged using the same procedure as for measuring the HP fluorescence decay, except that the emission monochromator is set equal to the excitation monochromator. The averaged fluorescence decay can then be obtained with the computer calculated convolution of the average time-profile of the scattered excitation with a suitable multi-exponential decay law. 4. Steps 2 and 3 are repeated for all the other HP concentrations. The best-fit decay tiine constants and relative pre-exponential factors are determined for all solutions.

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Guitar Techniques (March 2013)

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