I am trying to encapsulate hydrophobic doxorubicin in liposme, i can not see any absorbance peak for DOX, but there is a very sharp fluorescence peak of DOX,that indicate that DOX is encapsulated inside the liposome, Can anyone suggest what is the problem with absorbance peak?
You should not have fluorescence without absorbance! can you show spectra, and give details of measurement?
Yes Sadaf, as dox is a flourescent drug, you will have flouresence and absorption peaks. and you need to check the encaspulation efficiency right? If yes, use dialysis method and quantify the encapsulated drug by HPLC.
DOX is typically available as salt of HCl, and it should be soluble in water and alcohols. I remember about 50mg/mL stocks in water or buffers, more strengths in alcohols.
Any Fluorescent molecule will have absorbance, it cannot emit light without absorbing.
You need to review your methods in spectroscopy, DOX has absorbance at ~290 and ~480, with emission at ~580nm.
I had experienced this kind (I did not observe absorbance )while I was trying to encapsulate the DOX-Cl in a polymer I think it may be due to over dialysis or insufficient amount of DOX-Cl used or it could be due to inappropriate solvent selection
Can you check whether your DOX is present as microcrystalline aggregtes?
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