I extracted fungal extract (extra cellular ) with ethyl acetate . I want to do some bioassays for this.Now I want to check cytotoxic effect of this extract. which assay will be effective for crude extract? how is brine shrimp lethality assay?....do I need to dissolve extract in new solvent or ethyl acetate is ok?
Dear colleague, we have done a lot of Brine Shrimp assays over the years, and there are a couple of problems to consider:
1. Most importantly, make sure your brine shrimp are actually hatched in seawater. Many protocols simply indicate that you can make your own saltwater solution (e.g. using (NaCl), but this does not work well. Brine Shrimp in a regular salt solution are normally weakened, and die quickly even if you do not add an extract. The only way to be sure your Brine Shrimp die from your extract is using real (sterilized) seawater or a solution prepared based on seawater.
2. You need to either remove any solvent and dissolve your extract in the same solution the Brine Shrimp are hatched in, or run a pre-assay with in your case ethyl acetate, which will kill Brine Shrimp on itself, to determine lethality without your extract. However. it would be much better to remove the solvent and reconstitute in saltwater.
Dear colleague, we have done a lot of Brine Shrimp assays over the years, and there are a couple of problems to consider:
1. Most importantly, make sure your brine shrimp are actually hatched in seawater. Many protocols simply indicate that you can make your own saltwater solution (e.g. using (NaCl), but this does not work well. Brine Shrimp in a regular salt solution are normally weakened, and die quickly even if you do not add an extract. The only way to be sure your Brine Shrimp die from your extract is using real (sterilized) seawater or a solution prepared based on seawater.
2. You need to either remove any solvent and dissolve your extract in the same solution the Brine Shrimp are hatched in, or run a pre-assay with in your case ethyl acetate, which will kill Brine Shrimp on itself, to determine lethality without your extract. However. it would be much better to remove the solvent and reconstitute in saltwater.
U can choose brine shrimp lethality bioassay method for cyto-toxic evaluation of your extract. If u want to perform the assay method for crude extract ,i suggest u to dissolve your extract in water at 5mg/ml concentration..It may give u the effective cytotoxicity of crude extract. Ethyle acetate itself responsible to kill the brine shrimp..
You can prepare artificial sea water according to the method by Lymington and Fleming (1940) and hatch the eggs and dissolve the extracts in this solution.
Very effective if you use sea water that are not polluted and stick to the time allowed. Lab prepared salt solutions have failed many times
- Badges
- Science topic
- Similar topics
- Biological Science
- Plant Sciences
- Ethnobotany
- Natural Product Pharmacology