Chromatography Forum

I am currently using Shodex normal reversed phase column RsPak DE-413L column for HPLC analysis. The compounds that I intend to analyze is PHENOL, GUAICOL (2-methoxyphenol), and CATECHOL(1,2-dihydroxybenzene).
The following conditions were used for the analysis: flowrate 1.0 mL/min; eluent 0.005 M HClO4 aqueous solution/CH3CN = 90/10; oven temperature 40oC; UV/vis detector 220 nm.
However, i find that the RsPak De-413L is not able to separate these compounds (phenol, catechol and guaiacol) and when analyzed in HPLC, it will come out together as 1 peak. Is there anyway to make this compounds eluted separately in HPLC. I would like advice on this matter.

Did you try HILIC? It might help

Hi, Good afternoon.
Acidic phase, and too much water. Guajakol small soluble in water and sat down on the stationary phase. Try to wash column with pure methanol or methanol -wather 9:1 , and further use phase with 60-70% acetonitrile.

at what retention time do they come out as one peak?
is it at the front of the column?
or is it at a different retention time?
what is the pH of your aqueous solution?

depending on the answer to this you have several possibilities,

I think U should try the column which have more silanol activity. Try normal phase column with Hexane methanol combination. I think It will work rather than another combination.

Best regards

Praveen