I am planning to perform HPLC analysis of the degradation products of different dyes including Congo Red as well. But majority of papers didn't cite the temperature specifications of HPLC instrument oven. So if any one familiar with analysis of dye metabolite through HPLC please let me know that whether I need to keep the oven temperature of HPLC at 25 degree Celsius (i.e. room temperature) or there must be any specific temperature range for dyes?
Column temperature is a useful tool for lowering the system pressure in reverse phase HPLC through lowering the viscosity of the mobile phase. Use of higher temperatures can enable increased flow rates and mobile phase mixes that might not otherwise be possible at 25C because pressure becomes too great. Higher temperatures usually lead to components eluting more quickly which can also sharpen up peaks that have otherwise have relatively long retention times; on the flipside, at higher temperatures, there may be instances of peaks, which were resolved at 25C, coalescing owing to a reduction in retention time . However, when using simple solvent/water mixtures in RP HPLC, adjustment of temperature does not alter selectivity significantly, so I would not expect it be that helpful as a means of improving separation of dye metabolites. There is the issue of stability of dye metabolites at elevated temperature, but since typical upper temperatures tend to be no more than 40-50C, I wouldn't anticipate this as something to worry about. As someone who has performed a lot of analysis of dyes by HPLC, there is no specific temperature range required for colorants by RP HPLC. I suggest, to keep things simple at least for your initial studies, setting your oven temperature to 25C.
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