Hello All,
I will soon start to use HPLC and LCMS for 2 different studies. I have an animal tissue and a cell culture to be used for both the studies. The question that I have is what is the reliable/best method to follow while preparing samples for both the instruments? I have been getting a lot of materials on the interent but unsure of which to follow. Kindly suggest if there are any readily available kits also to prepare samples for the same.
Thanks
Surfactants can be applied to extract your cellular analytes but only neutral surfactant may be applied for LCMS because positive surfactant may not be applicable. Animal tissue uses laser desorption like MALDITOF, or thermospray LCMS which uses glycerol in mobile phase. I don't know what ionises DMSO buffer or what that takes.
Are you extracting from the cells or thinking of injecting cells through those instruments?. A flow cytometer is good if you label cells.
Rotavor or a rotavapour apparatus to remove crud from cells is good but getting cells soluble is a challenge itself. Maybe a strategy like coat cells with alginate and use a surfactant to bind alginate can keep them in solution. Organic solvents normally kill cells so using less cell density to check if in solution is GLP. Maybe nanoparticles bound to cell ligands can alter hydrophilicity. First off what mobile phase do you want to run in hplc because organic solvent kills cells such as 90 to 100% methanol. There should be papers on how to prepare mushroom extracts for analysis on cellular death.
hello Achintyan Gangadharan
it is depend on your aim and your samples , i will try to help you if you told me more.
I just used for HPLC a concentration betwen 1-5 mg/mL but depends of a lot of characteristics of your sample, and your method (reverse, SEC, cationic etc.)
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