Working with liposomal Amphotericin B i have tried many organic solvents like acetontrile, methanol, DMSO, Triton X but i am not able to get the complete digestion of liposomes for release of free amphotericin B what could be the reason, can anyone suggest.
Hi,
Chemical digestion of lipid vesicles in order to calculate %EE is problematic and can lead to erroneous results.
Better to separate the "free" (un-encapsulated) drug by physical means such as filtration or centrifugation.
Hi,
In my opinion,
Firstly, separate the free drug as mentioned by Dr Mozafari. Then, re-suspense the liposome and digest it with proper lysing agent and calculate the amount of unloaded drug. It depends more on the method you use for quantification of the unloaded drug. You can use surfactants such as TX-100, TW-20, with considerably low values or even methanol. Then, you can easily calculate Loading efficiency. The employed lysing agent should not interfere with the quantification approach or remove its interference by proper design of the measurement approach.
I hope that these tips will be useful to you.
I have optimized the approach for seperation of free drug and i am using the filtrate ( containing the liposomes associated drug) for the quantification using calibrators prepared from free drug, for the process of digesting the liposomes i have tried using methanol or surfactants like TX-100 but the liposomes specifically the one used for in ambisomes are not lysing to the maximu extent and i found it to deliver only 80-85% of free drug.
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