My sample is soluble only in toluene. The UV detection wavelength is 208 nm. But UV cut off for toluene is 285 nm. Can I use toluene just to dissolve samples? I have to do 5-10 microL injection. I do not have toluene is the mobile phase. I use acetonitrile, isopropanol and hexane.
I don't see any reason for not using toluene for dissolving your sample, but concern about whether or not it binds to the column too tight to be eluted. Expectedly toluene will be gone after injection and your UV provile won't be affected.
Thanks for your answe. I think toluene will not bind too tight in c-18 columns!
Zhao is right, your sample solvent is not a problem - it should be eluted quite early - I was thinking whether the HPLC solvent system you have mentioned would elute your sample latter on from the column as you mention it only dissolves in toluene - I wonder whether acetonitrile, isopropanol and hexane would elute it.
Yes, you are also right. But the sample is not completely insoluble in the mobile phase which I chose. There is some solubility at lower concentrations. I believe, the mobile phase at 1ml/min will elute it.
If your compound is soluble only in toluene it will precipitate in the column during the run analysis.
Thank you Dr. Massimo for your valuable comment. My sample is slightly soluble in the mobile phase (@ 20 mg/ml). Do you think it should be 100% soluble in mobile phase @ 10microL injection?
I think you can replace Tolune with a mix of Iso propyl alchohol with Acetonitrile to solubilize your compound.
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