I am using the hot phenol method to extract RNA. Usually, I am dissolving Phenol in water and keep it in the fridge ( 4°C ), i could see that it is a saturated solution because of the solid phenol on the bottom of the container.
But recently my solution gets "diluted" over time, the phenol in excess disapears and the extraction isn't as efficient as it was before.
Does anyone have an idea what might be the reason?
I think, keep the solution in cooling under nitrogen, may preserve the activity of solution, also in dark glass bottle
also, try to keep by freezing, then put in the hot water bath before your trial or experiments
It is preferable to prepare a phenol solution and use it instead of keeping it in refrigerator, which changes the concentration due to volatilization
Phenol in aqueous solution is quite prone to oxidation by exposure to air ― even air inside the closed container ― and sunlight; while the solute also tends to evaporate from the aqueous solution, particularly if exposed to open atmosphere. Moreover, water uptaked from humid air rapidly condenses and easily adds to the solution, if the solution container taken from the refrigerator is opened while still colder than the (laboratory) air dew point. You may considerer to close the container under blanketing inert gas; or otherwise the container should preferably be kept nearly filled. Photographers often use glass balls, or specialized squeezable plastic bottles, to protect phenol-based developer solutions from air.
Since phenol aqueous sol'n is prone to oxidation by air and light It is generally recommended to use fresh saturated phenol or it can be stored in dark brown bottle, tightly closed in frozen state.
About phenol oxidation ― also at RG: https://www.researchgate.net/post/Could_you_help_me_about_phenol_preparation https://www.researchgate.net/post/Phenol_having_a_reddish_Brown_color
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