I have used different reconstitution solvents in my sample extractions for an example, I used methanol as my reconstitution solvents in the water and sediment extraction sample whereas, I used acetone in my biota sample extraction. The results are all good, the compound peaks appeared symmetrically. However, the usage of acetone as my reconstitution solvent in my biota sample extraction seemed to be less efficient due to its high volatility even in my vial, the volume dropped from 1 mL to 0.5 mL over time.. In my biota sample extraction, I used hexane and acetone as my extraction solvents which the primary reason why I used acetone as my reconstitution solvents before injecting it in my HPLC. If I change the solvent:
1. Does it will affect my peaks in the HPLC?
2. If I were to change it to methanol, does the miscibility problem between hexane and methanol will occur?
Hope anyone can answer my questions. Thank you.
1. It depends on official mdthod
2. We can use polar and non polar solvent as methanol : hexane or methanol: acetonitrile
It would be good if you could test the miscibility of your sample in different solvents. If you want to quantify the compounds in that sample, the volatility of the acetone would be a problem. In general, it is recommended to reconstitute your sample in something compatible with your mobile phase. You can even make a mixture of solvents to modify the polarity.
Utilization of the different solvents and expected compounds identified should be analyzed in comparative appraisal. Also, some of the most important features of solvent and analyte (chemical and physical) such as density, polarity and non-polarity, refractive index, wavelength and so on should be considered.
Eshak Mourad El-Hadidy thank you so much for the answer, prof.
Gabriela Barragan I see, I'm currently using Acetonitrile: H20 for my mobile phase. So I think it's compatible with the reconstitution solvents that I'm using right now, acetone. But due to higher volatility, I'm changing it to methanol. the result are still the same.
Hamid Reza Farhang that's true. thank you for the suggestion! :)
That's right, methanol might work better because it's not as volatile. You just have to make sure that your sample is completely solubilized in methanol.
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