Hello Everyone, i'm actually using Shimadzu RF-6000 Spectrofluorophotometer and did not understand how to interpret the results and is it suitable to do a chlorophyll quantification?
Driss Ameziane Lhassani Have you seen any calibration in the literature? Usually, chlorophyll concentration is estimated using absorption spectroscopy, why do you want to use fluorescence?
Unfortunately, we do not have absorption spectroscopy in our laboratory, we have a spectrofluorometer (Shimadzu RF6000). So I ask if it is possible to quantify chlorophyll with this tool.
Few steps for the process. You can use the Beer-Lambert law (A=eCl) to generate a plot of absorbance of the dye as a function of the concentration. From the slope of the plot, you can calculate the molar absorptivity coefficient (e). Make sure you obtain a linear regression during this process. I would recommend you to stay stick to the absorbance range 0.1 to 0.9 for the best results. From the slope of the linear plot you will get the e value. Then you can test the sample of interest and measure the absorbance. If your absorbance reading is too high, make sure to do necessary dilutions. For the best results, I would again recommend you to maintain your sample dilution to record an absorbance value in the range 0.1 to 0.9. Once you recorded the absorbance, you can calculate the concertation from the equation A=eCl(where C is the concentration and l is the path length which is 1 cm in a standard cuvette). Make sure to consider any applicable dilution factors while calculating the concertation of you r sample.
Good Luck Driss Ameziane Lhassani !
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