I want to run HPLC for my Water extract. For my plant no paper found to chose a solvent system for HPLC.
Use ACN:Water in 80:20 combination. As there are no previous reportings, you will have to do hit and trial with many combinations to get the Rt in a usable range. Try many combinations to get the exact one for u. Also use methanol: water if this doesn't work.
Regards
Gud Luck
Thank you very much Dr. Alok. Yes I am not targeting any specific compound from my H2O extract. that's why I want to analyse the compound profile. Do you think to use reversed phase TLC to get the suitable combination of mobile phase?
Acetonitrile: methanol:95: 5 is useful in analysis of water extract.Prepare sample in HPLC water
To find a suitable solvent system for HPLC you MUST try it on TLC first. Usually Reverse phase column is good for water extracts. You have to, however, make sure that your solvent system elutes everything from your column. Otherwise, you will damage the column by leaving deposits in it. So try on reverse phased TLC plates and see which one elutes everything. In case that you get a good separation in one solvent system and complete elution in another you can use a gradient solvent system. Prepare your sample in water methanol mixture (preferably the ratio that you are going to use for elution in the beginning or the closest to it). Do not forget to filter it no matter how clear your solution is.
@Moyeenul Haq.. Yes it is a good idea to first perform a reverse phase TLC for getting the best solvent system. As reverse phase column is preferable for aqueous samples, it is good to go with it and try once for the better results. Find out the best solvent system and prepare your sample accordingly for analysis. It will no doubt take ur extra efforts, but if you succeed then lot of time will be saved later.
Regards
many thanks to Sandipan Datta and Dr Alok for your suggestions.
Dear Moyeenul Huq
is your sample plant or bacterial or animal extract because there are different solvent can used, anyway there is Acetonitrile : methanol in ratio 90:10.
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