Hi, I'm going to make a micropropagation and I want to add to my MS Media 2mg/L of activated charcoal as an antioxidant, but I want to know the best way to prepare the solution, I know that it's insoluble in water.
Thanks
add the finest grade activated charcoal last and then check the pH. after dissolving the gelling agent, stir the solution and pour it into the containers. autoclave and after autoclaving move the containers a little to avoid any eventual precipitation on the bottom of the coal. then let it cool. as activated carbon can cause the hydrolysis of saccorose and the lowering of the pH, if this phenomenon causes inconvenience to your culture, try to prepare a solution of water and coal separately, prepare your substrate with less water (the volume of the carbon to obtain the right diluition of your salts) and after autoclaving of both solutions pour one into the other, keep stirred and then under a sterile hood fill your containers
In tissue culture, it is added as a powder in the last step of preparing the nutrient medium before adding the agar
Hello Julia,
thank for posting this interesting technical question on RG. Activated charcoal is a form of (impure) carbon with a high surface area. To the best of my knowledge, activated charcoal is not only insoluble in water but also in all organic solvents. Thus (unfortunately) the answer to your question is that you cannot prepare an activated charcoal solution. You can either use it in suspension of add it as finely powdered solid in the desired amount.
P.S. It might be possible to "dissolve" activated charcoal in so-called piranha solution (i.e. a mixture of concentrated sulfuric acid and hydrogen peroxide). However, once it is dissolved, it isn't activated charcoal any more....
Good luck with your work and best wishes!
Dear Julia
I agree Alberto, activated Charcoal ( 2 g) is added in the form of a fine powder to the medium (liter) immediately before adjusting the pH with continuous stirring. Also, during pouring the medium into the tubes/flasks with stirring, finally after autoclaving you must shake the tubes /flasks manually or on shaker before solidified (to prevent the coal to participate). I think this method suitable with a least damage to apply for micro-propagation.
Note: weight of activated charcoal has to adjust according to the volume of medium...................good luck.
regards,
Metwally bekhit
An activated charcoal is solid insoluble in water and other solvent, hence it is difficult to get its solution.
Julia Vasquez Add 2 mg of activated charcoal powder (per Litre medium with optimized hormones etc.) just prior to autoclaving. Stir well and pour into culture vessels. It use to be suspended in the medium and moreover two types of AC are used in PTC such as acid-washed and neutralized AC.
Julia Vasquez, Activated charcoal in the concentration of 2mg/l is a very low concentration for plant tissue culture. So, I think you mean you want to prepare an MS medium with 2 g/l activated charcoal. Anyway, it's better to heat the medium till about 80 °C on a heater and stirrer, then add the AC and wait for 5 min. to have a homogenized suspension. Then you can pour the medium into the culture vessels/jars while shaking it manually. After autoclaving, the culture vessels need to be shaken to avoid AC deposition.
Of course 2 mg/L of AC would be less, but 2 g/L AC is more. In general we are using 100-200 mg AC per litre of nutrient medium.
you can make media with AC, Grogel, MS/ Gamborgs media, sugar and distilled water maintain pH 5.8 . finally autoclave them.
After dissloving agar completely through heating, Activated charcoal of desired percentage can be added in the medium and dissolve over the flame. The ac should be completely dissolved so that medium should looked uniformly colored when solidified.
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