I am doing fermentation based screening of novel secondary metabolites from Bacteria. Which solvent i should use for HPLC and MS analysis. Should I use a single or different set of solvents like, Ethyl acetate, N-hexane, Chloroform etc . to do these analysis HPLC, MS.? can i Use a the cell free supernatent directly for HPLC or MS meabolites?? Kindly need your valuable suggestions.
There are many factors (such as solubility, polarity, stationary phase and ......) you should consider. I think that optimal way is following previous publications which have similarity to your work.
If you meant using direct crude extract after extraction to inject into HPLC, it is usually not done because of the possibility of spoiling the chromatographic column. Purification and follow-up are sought to see if he continues to extract what he is interested in, but look for works that have drawn his target of interest
I would suggest that first of all you do specify your desire secondary metabolites, purify it then you can find best mobile face because it entirely depends on the solubility of your secondary metabolites. If you go directly for HPLC of your crude metabolites then firstly, you will extremely damage your HPLC colomn, secondly, it would be very difficult for you to recognize and identify the resultant peaks after performing HPLC because of having hundreds of peaks and thirdly, you will loss many important compound because of their insolubility in your mobile phase.
Noor Hassan thanks for your kind suggestions. However, I am more interested to know the complete picture of the 2ndary metabolites the micrbes produces Then, I will cross check with and Transcriptomics and Bioinformatics analysis to find the week or Silent biosynthetic gene cluster. Furhther i want to activate the silent or weak gene clusters using CRISPR/CAS9 system,
I am trying it with Different solvents. I am hoping for some novel compounds.
- Badges
- Science topic