I am working with Fluorescence spectrophotometer (Model:F-2700 FL) to determine concentration of chlorophyll a and carotenoids in cyanobacteria. I set up the excitation and emission wavelength of beta carotene as 425 nm and 520 nm respectively. While chorophyll an excitation and emission wavelength were set at 430 nm and 680 nm respectively. By running the analysis, a longer and a shorter peaks appeared as could be seen in the picture. Each time i changed the excitation and emission wavelength in order to determine other pigments such as chlorophyll a, lutein, astaxanthin e.t.c i got similar peaks. Please how can i identify a particular pigment and its concentration from the spectral peaks? why the peaks remain same each time i run the test despite variation in the wavelength? Beside HPLC, can i use the fluorescence machine to characterize carotenoids composition? Thanks.
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