Thursday, July 16, 2009

Prepratio of Trish HCl Buffer






Follow the steps
1. Calculate Moles of Tris Base
Determine the number of moles of Tris base needed by multiplying the desired molar concentration of buffer by the volume of buffer being made.
mol/L * L = moles needed {For Eg. you need 1M 100ml trish buffer solution}
2. Calculate Mass of Tris Base
Determine the mass of Tris base to weigh by multiplying the number of moles by the molecular weight (121.14 g/mol) of Tris.
Molarity = {Wight of Trish * 1000} / molecular wt of Trish
[ Eg. 1M = {Wight of Trish * 1000} / 121.14 ;
{Wight of Trish * 1000} = 121.14 ]
3. Dissolve Tris Base in Water
Dissolve the required mass of Tris into a volume of deionized water approximately 1/3 of the desired volume of buffer to be made.
4. Adjust the pH
Using a pH meter, titrate the solution of Tris with 1M hydrochloric acid (HCl) until the correct pH is reached.
5. Bring to Volume
Add the TrisHCl mixture to a volumetric flask of the desired volume and add deionized water as required to complete the solution.

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