Preparing a 700mL PBS Buffer Solution: Step-by-Step Guide

  • Thread starter Thread starter WSR
  • Start date Start date
Click For Summary
To prepare 700 mL of PBS at pH 7.40 with 25 mM phosphate and 140 mM NaCl, use the Henderson-Hasselbalch equation to determine the required concentrations of Na2HPO4 and NaH2PO4. The total concentration of hydrogen and dihydrogen phosphates must equal 25 mM, allowing for the calculation of individual concentrations. After determining these concentrations, calculate the volumes needed from the 1 M stock solutions to achieve the desired final volume. For the second question regarding pH and protonation, the approach using the Henderson-Hasselbalch equation is correct, and the calculated pH of 7.89 aligns with the given pKa. Accurate calculations and dilutions will yield the desired buffer solution.
WSR
Messages
1
Reaction score
0
Question

You are asked to prepare 700 mL of PBS (phosphate buffered saline). The PBS needs to be pH 7.40 with 25mM phosphate and 140 mM NaCl. You are given a bottle of NaCl (58.45 g/mol), 1 M Na2HPO4 stock solution and a 1.00 M NaH2PO4 stock solution. ( H3PO4 pk1=2.12, pk2=7.21, pk3=12.66) Describe how to prepare this buffer.


Equations

I'll use the Henderson-Haeelbalch Equation so pH= pka +log(base/acid)


The thing that really screws me up in this problem is how to incorporate the M concentrations. The pH has to be 7.40 so 7.40 = pka + log (base/acid). I don't know where to go with the concentrations.


Also, a separate question that I don't know if I did correctly.
At what pH will 62% of x be in the protonated form if pka = 8.10. So I did, pH= pka +log(base/acid) and got pH= 8.10 + log (.38/.62) with an answer of 7.89. Does that makes sense?

I'd appreciate any guidance.

Thanks
 
Physics news on Phys.org
You know sum of concentrations of hydrogen and dihydrogen phosphates must be 25 mM. That, plus Henderson-Hasselbalch equation give you two equations in two unknowns - solve for both concentrations. Then calculate how much of the original solutions you need to dilute to produce 700 mL of the PBS.

Approach to the other question looks OK, I haven't checked numbers.
 

Similar threads

  • · Replies 1 ·
Replies
1
Views
4K
  • · Replies 3 ·
Replies
3
Views
3K
Replies
3
Views
4K
  • · Replies 7 ·
Replies
7
Views
3K
  • · Replies 7 ·
Replies
7
Views
3K
  • · Replies 23 ·
Replies
23
Views
3K
  • · Replies 11 ·
Replies
11
Views
3K
  • · Replies 1 ·
Replies
1
Views
3K
  • · Replies 2 ·
Replies
2
Views
10K
  • · Replies 1 ·
Replies
1
Views
2K