Hi Guys,
I tried to filter my cubosomes nanoparticles to do cytotoxicity testing. However, only 500 uL of nanoparticles was obtained upon filtration of 1 mL of the nanoparticles (Corning filter, 0.2 um).
I strongly suspecting the nanoparticles concentration will be affected as some might be block by filter (Theoretically average size around 190 nm), and the concentration is not accurate anymore due to lower volume which I obtained.
Are there any methods to test the particles concentration after the filtration? Would like to understand the nanoparticles recovery after the filtration...
Or any other methods recommended to do this?
Thanks!
Hi
your solution that your nano-particles dissolved it , is hydrophobic or hydrophilic ?
if it is hydrophobic you used hydrophobic filter.
if you want test the particles concentration after the filtration, you can used from spectrometer before and after filtration
if your average particles size around 190nm and you have high PDI value, then there is high potential that you have particles more than 220 nm which will be trapped in the fillter.
if your particles are stable thenn you can do sterilisation with other method such as heat or autoclace.
if you are using lipid based particles, then I would suggest to extrude your particles with a 100 nm PC membrane to reduce the size and then steralise them with filitration.
there are different method to analyse the concentration of your particles, you can use HPLC or if your formulation contains cholesterol then you can use cholesterol detection kit.
see this paper for HPLC analysis:
http://www.sciencedirect.com/science/article/pii/S0378517316310675
Hi Obeid,
Thanks for your suggestion. However, my lipid nanoparticles are phytantriol cubosomes with high PDI value. How do I quantify it using HPLC as I strongly suspecting there will be multi-peaks appearance due to their heterogeneous size? No cholesterol/presence of other monodisperse particles inside the cubosomes system for me to make the quantification...
High PDI value means poor stability and high variablity in drug encapsulation and cellular uptake, I would recomend to do size reduction first by extrusion to make your PDI less than 0.2.
Analysis with HPLC depends on the lipid components not the size, I dont have much expereince with cubosomes but you can dissolve the particles with 100% isopropyl alcohol to rupture the particles and then analyse how much you have from each lipid component by HPLC or any other method of analysis.
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