I am a little bit confused about Tris-HNO3 buffer (pH 8.2, 1%NaCl). I have only HNO3 and NaCl reagent available. However, Tris and buffer words make me confused. In this case, I need to be more clarification about the preparation of Tris–HNO3 buffer (pH 8.2,1%NaCl) from HNO3 and NaCl reagent.
Hi Abul,
you need to obtain Tris Base powder (known by some suppliers as TRIZMA) as the third ingredient to provide the buffering potential. Please note that this is not the same as Tris HCl (which has already been acidified with HCl).
For instance, from Tris Base from Sigma : http://www.sigmaaldrich.com/catalog/product/sigma/t1503?lang=en®ion=GB
If you are making (for instance) 1L of 0.1M Tris- buffer (HNO3- acidified), 1% w/v NaCl pH8.2, then you would require :
12.1g Tris base (=100mM)
10g NaCl (1% w/v, = 171mM)
Dissolve these in ~800ml of sterile water. Then, adjust pH to 8.2 using HNO3. Finally, adjust volume to 1L (and verify pH has remained constant).
Good luck.
I will just add one thing. When you add nitric acid to the Tris buffer solution, the temperature will increase. The pH of Tris buffer is temperature dependent, so you will have to allow the buffer to cool back down to the desired temperature before completing the pH adjustment.
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