# How ph affects aesculune fluorescence?

I am facing a ween problem. I am trying to find out, how would look like the graph for fluroscence of aesculine (intensity of fluoroscence on Y axis and pH on X axis).

I know, that increasing the pH causes a corresponding increase in the maximum fluorescence intensity from 77.74 (units) in the acidic sample to 146.13 at neutral pH and 232.69 at alkaline pH.

But, can I apply this on aesculine?

• You should measure the fluorescence vs that from a a standard compound excited at the same wavelength (with same absorbance) and observed at the same wavelength. If you look in the literature there are many standards and also compounds whose fluorescence yields are known. Look also for measuring relative fluorescence yields rather than absolute yields as these are v tricky. In any experiment you must ensure that you correct for any differences in the amount of light absorbed as this can be a source of large errors. – porphyrin Mar 22 at 16:22
• A pH dependence is an indication that a pH buffer should be used in the sample matrix before the instrumental analysis. – MaxW Mar 22 at 17:11
• I have no idea, you are talking about... – HASHTAG Mar 22 at 17:26
• Keep also in mind that coumarins are labile at $\mathrm{pH} \ge 12$. – Mathew Mahindaratne Mar 22 at 17:50