Is it correct to filter the extracts we obtain with Whatman filter paper and use them directly in experiments to investigate the bioactivity of phytochemical substances? Or is it correct to first centrifuge the extracts we obtain, then filter the supernatant and use them in experiments?
It depends a bit on the experimental setup you purposed. For instance, if an enzyme inhibitory activity of the phytochemical extract is the subject for investigation, no problem for the use of filter paper only before application. For the in-vitro assays such as cell culture experiment, I prefer to clarify extract applying both centrifugation and filtration prior to spiking...
It is based on the nature of your extracts and the desired level of clarification for downstream assays
Choose the method or combination of methods that best balances clarification with biomolecule preservation for your specific needs.
If you are doing only filter paper (i.e. rough) filtration then you probably don't need centrifugation.
If you are going for fine filtration (e.g. 0.45/0.22um) you are likely to clog your filter very fast. Pre-centrifugation usually helps to avoid such.
Either way, remember that filtration is separating by size while centrifugation is separating by density, and they don't have to be the same! Think straw floating on water
Hello Avni Yildizbaş
You should first carry out a rough clarification of the extract by simply decanting, followed by a filtration step by using a six to seven layered muslin cloth. Then you may include a centrifugation step which may be required if the powder is too fine to be filtered.
I have attached a book below which may be helpful.
Article Methods in Biotechnology. Natural Products Isolation, Second...
Best.
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