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This optical absorption measurement of Phenylalanine were made by R.-C. A. Fuh in the summer of 1995 using a Cary 3. The absorption values were collected using a spectral bandwidth of 1.0 nm, a signal averaging time of 0.133 sec, a data interval of 0.25 nm, and a scan rate of 112.5 nm/min.
These measurements were scaled to make the molar extinction coefficient match the value of 195cm-1/M at 257.5nm (Fasman, 1976).
The fluorescence emission spectrum of Phenylalanine dissolved in water. The excitation wavelength was 240nm. The quantum yield of this molecule is 0.022 (Chen, 1972). This spectrum was collected by in the summer of 1995 using a Spex FluoroMax. The excitation and emission monochromators were set at 1 mm, giving a spectral bandwidth of 4.25 nm. The data interval was 0.5 nm and the integration time was 2.0 sec.
Samples were prepared in 1cm pathlength quartz cells with absorbance less than 0.1 at the excitation and all emission wavelengths to uniformly illuminate across the sample, and to avoid the inner-filter effect. The dark counts were subtracted and the spectra were corrected for wavelength-dependent instrument sensitivity.
Chen, R. F. (1972) Measurements of absolute values in biochemical fluorescence spectroscopy. J. Research National Bureau Standards 76A(6), 593-606. The same article appeared in Accuracy in Spectrophotometry and Luminescence Measurements, Nat. Bureau of Standards Special Publication 378, 183-196 (1973).
Dixon, J. M., M. Taniguchi and J. S. Lindsey (2005), "PhotochemCAD 2. A Refined Program with Accompanying Spectral Databases for Photochemical Calculations, Photochem. Photobiol., 81, 212-213.
Du, H., R.-C. A. Fuh, J. Li, L. A. Corkan and J. S. Lindsey (1998) PhotochemCAD: A computer-aided design and research tool in photochemistry. Photochem. Photobiol. 68, 141-142.
Fasman, G. D., Editor (1976) Handbook of Biochemistry and Molecular Biology, 3rd Edition, Proteins, Volume I, pp. 183-203, CRC Press, Cleveland, Ohio.