My work involves the production of melatonin using saccharomyces cerevisiae and its quantification. I have tried various fluorescence spectroscopic methods but couldn't find an optimized protocol for it. Kindly help me with the same
I think this will be very challenging because there will be a high background of fluorescence from other molecules with similar fluorescence spectra, including most proteins, due to their containing tryptophan residues.
Probably the best ways to quantify melatonin in a cell extract are (1) an immunological assay such as ELISA, and (2) liquid chromatography, combined with mass spectrometry (LC-MS). In both cases, you need a standard sample of melatonin to prepare a standard curve.