I need to find the concentration (in mol/L) of the PEG-Hematin I synthesized. The extinction coefficient is 70000 mol-1cm-1 obtained from the calibration curve of the hematin sample in DMF. The 400 nm peak has an absorbance of 1.25 when taken from a solution prepared by dissolving 0.003 g of PEG-Hematin in 3 mL DMF (1 g/L). Any help to find the hematin concentration would be appreciated. Thanks
Oh, of course. Thank You. The value of 1.85 absorbance I used is actually that of 395 nm, not 280 nm. I assume that means the concentration I found is of Hematin in my compound. How can I use this to find the concentration (in mg/ml or Molarity) of the PEG-Hematin in my product?
Or is my UV-VIS not helpful even for the calculation of Hematin because it's too concentrated?
I estimate the absorbance at the 400 nm peak to be about 1.25. Using the aforementioned extinction coefficient of 70000 M-1cm-1 gives a concentration of 17.8 µM. This assumes that there is no contribution from the PEG (which is colorless), or any artifact from light scattering by particulates. The molecular mass of hematin is 652 g/mol, so the hematin concentration is 0.0116 g/L. Your solution of PEG-hematin was 1 g/L. Therefore, the hematin makes up about 1.16% of the mass of the sample.
By the way, you said you were making a modified protein, but hematin is not a protein.
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