am working on analyzing the amount of HMF I have in the organic phase I use for my reaction( The organic phase is MIBK and 2-Butanol). I am currently doing low-pressure gradient. Find below more information concerning the mobile phase I am using.
Mobile phase A: 2.5mMsulfuric acid
B: ACN: Water 20% V/V.
Flow rate; 0.3ml/min
Injection volume: 5ul
I currently see a baseline drift at about 19min. I have tried to get it out but to no avail. Could you please suggest what I can do? Please find attached the chromatogram i current have.
Eliminate sulfuric acid, replace with TFA at the concentration of desired pH in both mobile phases. Use ACN or better MeOH, inject 1-5 mcL sample, After the peak of interest you can ramp up the gradient to clean any junk in the column if needed.
I am also working on determination of HMF content in honey by HPLC.
What is your sample matrix?
Anita Sood I am producing HMf from glucose. I am trying to analyze the amount of HMF in MIBK and 2-Butanol.
Andrei Blasko, you think if I use TFA I will get better result? I am currently using ACN: Water 20% V/V as my organic mobile phase and 2.5mMsulfuric acid. Do you also suggest i add some of the acid to my organic mobile phase?
You generally want to keep the pH constant in a gradient. Make up your channel B with the same acid concentration as the A channel. I am not a fan of sulfuric acid in the mobile phase. If it is compatible with your sample use phosphoric acid.
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