Pauline Bianca R. Alfonso December 3, 2012 Exercise 1 Buffers 1.

As a group, design and prepare a buffer according to the specifications provided by your instructor. Show your calculations below. Do not use a pH meter in your preparation of the buffer. Prepare a liter of 0.1 M of formate buffer at pH 4.0. This is a single-component buffer system. This means that one component of the conjugate acid/base pair will be generated in situ (in solution). In the following example, the conjugate base will be generated by reaction between the weak acid (acetic acid) and strong base (sodium hydroxide). a.) Use the Henderson Hasselbalch equation to find the ratio of to . [ ] [ ] [ ] [ ] [ ] [ ] [ ] [ ] b.) Calculate the decimal fraction (part/whole) of each buffer component.

c.) Find the molarity (M) of each component in the buffer by simply multiplying the molarity of the buffer by the decimal fraction of each component. [ ] [ ] d.) Calculate the moles of each component in the buffer. Moles = Molarity x Liters of buffer

e.) Note: Since this buffer is prepared by the reaction of a weak acid ( ) with a strong base (NaOH), you must determine the total moles of the weak acid component needed. This sum indicates that, although in the buffer one only needs an additional is needed to generate the conjugate base in solution. f.) Calculate the mass of NaOH and volume of weak acid required to make the buffer. ,

g.) Prepare this buffer, one would use appropriately sized pipets or cylinders to measure and transfer each component to a 1000 mL volumetric flask and bring the solution to volume with distilled water. This is a two-component buffer system meaning that the weak acid and its conjugate base are added separately. a.) Use the Henderson Hasselbalch equation to find the ratio of to . [ ] [ ] [ ] [ ] [ ] [ ] [ ] [ ] b.) Calculate the decimal fraction (part/whole) of each buffer component.

c.) Find the molarity (M) of each component in the buffer by simply multiplying the molarity of the buffer by the decimal fraction of each component. [ ] [ ] d.) Calculate the moles of each component in the buffer. Moles = Molarity x Liters of buffer

e.) Calculate the volume of formic acid and the mass of sodium formate required in the preparation of 0.1M buffer at pH 4.0.

2. Prepare your buffer. Report your experimental procedure below (give the exact amounts and glassware used). Do not use a pH meter in your preparation. Procedure for preparing your buffer: For single-component buffer system a) Add 10ml of water in 1000ml volumetric flask using 10ml pipet. b) Put exactly 1.38ml of formic acid in the flask using appropriately sized pipet. c) Weigh 2.54 grams of sodium hydroxide in analytical balance. d) Transfer it in a beaker. Dissolve it with 20ml distilled water. e) Pour this solution carefully to the flask that contains formic acid. f) Dilute the solution using distilled water upto the mark. Make sure that you mix the solution upon diluting.

 a.) b.) c.) d.) e.) f.)

For two-component buffer system Add 10ml of water in 1000ml volumetric flask using 10ml pipet. Put exactly 1.38ml of formic acid in the flask using appropriately sized pipet. Weigh 4.32 grams of sodium formate in analytical balance. Transfer it in a beaker. Dissolve it with 20ml distilled water. Pour this solution carefully to the flask that contains formic acid. Dilute the solution using distilled water upto the mark. Make sure that you mix the solution upon diluting.

3. Given the following problems, make a table showing pH vs [CH3COOH]/[CH3COO-]. Calculate pH when a) [CH3COOH]/[CH3COO-] = 0.1 b) [CH3COOH]/[CH3COO-] = 100 c) [CH3COOH]/[CH3COO-] = 0.01 d) [CH3COO-]/[CH3COOH] = 100 e) [CH3COOH] = 0.04M and [CH3COO-] = 0.08M f) [CH3COOH] = 0.39M and [CH3COO-] = 0.78M g) [CH3COOH] = 0.08M and [CH3COO-] = 0.04M pH [CH3COOH]/[CH3COO-] 5.75 0.1 2.75 100 6.75 0.01 6.75 0.01 5.05 0.5 5.05 0.5 4.45 2 a) b) c) d) e) f) g)
[ ] [ [ [ [ [ [ [ [ [ [ [ ] ] [ ] [ ] ] ] ] ] ] ] ] ] ]

Did you observe any pattern? What conclusion can you make? Yes. If there is a greater amount of acid, the pH decreases making it more acidic. Thus, the pH of a buffer is dependent on the mole ratio of the acid and its conjugate base. If [CH3COOH] > [CH3COO-], is the pH > pKa, or < pKa? If [CH3COOH] > [CH3COO-], the pH is less than the pKa. This is because the mole ratio of conjugate base to acid is less than is less than one resulting to a negative value when taking its logarithmic value.

If the total acetic acid added is 0.10M, what are [CH3COOH] and [CH3COO-] when the ratio is 1:1? If the total acetic acid added is 0.10M, the concentration of acetic acid and its conjugate baseis both 0.05M when the ratio is 1:1. If H+ is added until the ratio is 9:1 (high acid). What are [CH3COOH] and [CH3COO-]? If the H+ is added until the ratio is 9:1 (high acid), [CH3COOH] is 0.09M and [CH3COO-] is 0.01M. 4. a) b) A table of common buffers used in biology is attached. Answer each question. Choose a buffer that would be very effective at pH 3.8. Formate buffer How would you make the buffer system, at, say 0.1M? [What chemical(s) would you use? How much (in M)?] The buffer at its pKa is an equal mix of formic acid and formate ions. This can be done by mixing 0.052M of sodium formate to 0.048M formic acid or by adding a strong base such as sodium hydroxide to 0.1M formic acid until pH approaches near equivalence. [ ] [ ] [ ] [ ] [ ] [ ] [ ] [ ]

c) d)

e)

] [ ] You want to buffer a solution at pH 9.2. Your choice of buffer? Borate buffer Would Tris buffer be suitable for use at pH 7.0? The pKa of Tris buffer is 8.30 which is higher than the pH 7.0 by more than one (8.30 7.0 = 1.3). This means there is a high ratio of [HA]/[A]. Thus, there is a very little [A] left and has a little ability to buffer against base. One of the important buffers in your blood is based on phosphoric acid. Blood pH is about 7.4. Which step of the phosphoric acid ionization is most relevant in this situation? Write the chemical equation for this ionization. The second ionization of phosphoric acid is the most relevant in this situation. The chemical equation for this ionization is: H2PO4 H+ + HPO4-2

5) Describe the preparation of a liter of 0.2M acetate buffer. Determine the volume of acetic acid and sodium acetate solution that has to be mixed to obtain a 50.0mL buffer solution with the following pH. Fill the table below. Show your solution.

pH 3.6 3.7 3.8 3.9 4.0 4.1 4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 5.0 5.1 5.2 5.3 5.4 5.5 5.6 At pH 3.6 [ [

Volume of Acetic Acid solution

Volume of Sodium Acetate solution

] ]

[ [

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At pH 3.7 [ [ ] ]

[ [

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At pH 3.8 [ [ ] ]

[ [

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At pH 3.9 [ [ ] ]

[ [

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At pH 4.0

[ [

] ]

[ [

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At pH 4.1 [ [ ] ]

[ [

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At pH 4.2 [ [ ] ]

[ [

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At pH 4.3

[ [

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[ [

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At pH 4.4 [ [ ] ]

[ [

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At pH 4.5 [ [ ] ]

[ [

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At pH 4.6 [ [ ] ]

[ [

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At pH 4.7 [ [ ] ]

[ [

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At pH 4.8 [ [ ] ]

[ [

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At pH 4.9 [ [ ] ]

[ [

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At pH 5.0 [ [ ] ]

[ [

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At pH 5.1 [ [ ] ]

[ [

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At pH 5.2 [ [ ] ]

[ [

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At pH 5.3 [ [ ] ]

[ [

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At pH 5.4 [ [ ] ]

[ [

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At pH 5.5 [ [ ] ]

[ [

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At pH 5.6 [ [ ] ]

[ [

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6) Scenario: Your group is made up of sailors on a submarine that is on a secret mission. The doctor on the submarine went into cardiac arrest and he now has acute acidosis (which means his blood pH is too low). The pH of normal, healthy blood is approximately 7.40. Your submarine cannot surface to transfer him to a medical facility because your secret mission would be compromised. Since hes the doctor, youll need to save him without any professional help. You must rely on your knowledge of buffers. Suppliers on the submarine are limited and the only available pH meter is broken!

Procedure: Assumed that the buffer that you designed and prepared in step 2 above is the blood of the doctor after his cardiac arrest. Your job is to decide how you are going to raise the pH of this blood sample to a normal pH level. After you are confident in your calculations, carry out your pH raising procedure on the blood sample. Do not use the pH meter. Record your calculations and procedure below. Hint: If you need to add small amounts of some solution, consider using a small plastic dropper. You may need to calibrate the dropper so you know how many millilitres are contained in one drop. Other pipets are also available. Show your calculations. Devise a procedure: From the results obtained in number 1 (using 0.1M formate buffer at pH 4.0): Moles of sodium formate = Moles of formic acid = 0.036496 moles HCOOH + OH HCOO + H2 O 0.063504moles x 0.036496moles 0 -x -x +x [ ] [ ] [ ] [ ] [ ] [ ] [ ] [ ]