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Phosphoric acid (H3PO4) is a triprotic acid with the following pKa values: pKa1

ID: 1049910 • Letter: P

Question

Phosphoric acid (H3PO4) is a triprotic acid with the following pKa values: pKa1 = 2.15 pKa2 = 7.20 pKa3 = 12.35 You wish to prepare a 1.000 L of a 0.0450 M phosphate buffer at pH 6.8. Which of the following salts do you use (PICK 2) and what mass of each salt will you add to the mixture? H3PO4 (M.W.= 97.995) KH2PO4 (M.W.= 136.086) Na2HPO4 (M.W.=141.959) AlPO4 (M.W.=121.953) A concentrated phosphate buffer known as "phosphate buffered saline" (or PBS) is made with the salts you choose from above in addition to NaCl and KCl. The 10x concentrated solution has a pH ~ 6.8 and the 1x concentrated (i.e., diluted with water) solution is ~7.1. Why? You accidently grabbed the 10x concentrated phosphate buffered saline (or 10xPBS) instead of the 1 times PBS solution and start to adjust the pH of your buffer with NaOH to reach your desired buffer pH. You notice it is taking much more base than with your usual 1 times PBS buffer. Why?

Explanation / Answer

a)

if you want pH = 6.8 approx.. the best fit is the 2nd ionization point

pKa2 = 7.20

apply

pH = pKa + log(Na2HPO4 / NaH2PO4)

solve for 1 l

6.8 = 7.21 + log(Na2HPO4 / NaH2PO4)

10^(6.8-7.21) = 0.3890

Na2HPO4 = 0.3890* NaH2PO4

NaH2PO4 + NaH2PO4 = 0.045

substitute

0.3890* NaH2PO4 +  NaH2PO4 = 0.045

1.3890NaH2PO4 = 0.045

KH2PO4 = 0.045/1.3890 = 0.032397 mol

Na2HPO4 = 0.3890* NaH2PO4 = 0.3890*0.032397 = 0.012602 mol

mass of KH2PO4 = mol*MW = 0.032397 *136 = 4.4059 g

mass of Na2HPO4 = 0.012602 *141.96 = 1.7889g

so..

b)

this is mainly due to the activity/inic force of the acids/conjguate ions interactng between each other

c)

the ratios are the same, but actual amounts of acid/base are larger, so you require more base/acid respectively to react with the buffer