I need a dye that wouldn't interact with the ascorbic acid to be encapsulated as a marker for measuring the encapsulation efficiency using the UV spectrometer. I have tired to use some dyes but it was reduced by the ascorbic acid losing its colour.
I'm using DSPC:Chol in ethanol and the ascorbic acid in PBS 7.4. (micro fluidics method)
Any suggestions of a suitable dye??
Why not just use the absorbance of ascorbic acid itself? At pH 7.4, the absorbance maximum is at 265 nm. To determine the amount of ascorbic acid in the liposome sample, you would have to first dissolve the liposomes in organic solvent or detergent, because they would otherwise scatter the UV light and give a false signal. The solvent or detergent itself would, of course, have to be free of absorbance at 265 nm.
Adam is right, why not use the AA absorption value itself. You can also check this: A Chemiluminescence Reaction Between Hydroxyl Radical and Rh...
Adam B Shapiro Riaz A. Khan Thank you so much for your answers. I'll be measuring the liposomal ascorbic acid alone.
But my next step is entrapping another drug with the ascorbic acid and I would need to get an idea about the effect of ascorbic acid on the entrapment efficiency of the other drug. So I was thinking of using a dye as a mock drug where I can separate it with the sephadex G-50 then measure the absorbance..etc
I used the methylene blue but it was reduced by the ascorbic and lost the colour. I'm currently thinking of the crystal violet. Do you have a suggestion for that ?
If you expect the encapsulation of one drug to affect the encapsulation of a second one, then I recommend performing the analysis with the actual drugs, not surrogates, because different compounds may have different effects.
If you have access to an HPLC or UPLC system, you can use it to measure the amounts of each drug in samples of solvent-dissolved liposomes by separating them and quantifying the peak areas.
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