Titration of Amino Acids

Introduction

1 M glycine solution y 0.1 M aspartic acid solution y Neutralized formaldehyde y .Materials 0.1 N NaOH y 0.1 M lysine solution y 0.1 N HCl y 0.

introduce 10.0 mL of the amino acid solution and add 10mL of distilled water and measure the resulting pH of the solution 3.Procedures Take two pipettes and fill the first with 0. Into each of the two beakers. Titrate the first solution with 0. until a total of 10. .1N HCl adding 2.0 mL at a time and determining the pH after each addition.1N HCl and the second with 0.1N NaOH 2. measure the pH at 5mL and 15mL volumes 1.0 mL (for lysine).0 mL is reached (for glycine and aspartic acid) or 20. In addition.

1 mEq of acid/base. (Show how this value was obtained). The equivalent acid/base (abscissa). 0. Plot the pH (ordinate) vs. 6. Titrate the solutions.1N NaOH. Titrate the second solution in the same manner as the 1st using instead.0 mL is reached (for glycine and lysine) or 20. until 10.Procedures 4. One mL of acid/base = 0.0mL of neutralized formaldehyde solution into each of the amino acid solutions before determining the pH of the solution. Repeat the above procedure. but add 5. .0 mL is reached (for aspartic acid) 5.

Construct your titration curves on graphing paper. the equivalent acid/base on the same graph as above. Plot pH vs. 8.Procedures 7. Solve for the pI and pK values of your amino acid. Record your results as follows .

99 NaOH Gly w/o HCHO 6.98 10.28 9.06 6.99 5.33 2.92 7.58 NaOH Gly w/ HCHO 4.57 6.84 8.52 2.75 9.88 .91 9.21 7.62 2.96 2.02 2.35 2.57 6.56 2.97 2.13 1.15 2.69 9.43 2.Glycine: Glycine: Acid VS Base Data HCl Gly w/o HCl Gly w/ HCHO HCHO 0 2 4 5 6 8 10 5.26 10.12 1.

Glycine: Glycine: Acid VS Base Graph 12 10 8 6 4 2 0 0 2 4 HCl Gly w/ HCHO 5 6 NaOH Gly w/o HCHO 8 10 HCl Gly w/o HCHO NaOH Gly w/ HCHO .

78 12.45 2.64 3.16 11.58 12.81 .11 2.14 NaOH Asp w/o HCHO 4.5 4.75 2.99 12.2 9.34 12.3 2.64 3.47 3.43 10.63 11.Aspartic Acid: Acid VS Base Data 0 2 4 5 6 8 10 12 14 15 16 18 20 HCl Asp w/o HCl Asp w/ HCHO HCHO 4.91 1.62 7.64 3.18 8.32 12.22 2.9 9.84 1.04 1.03 1.47 3.47 4.01 3.12 9.5 4.68 11.53 10.96 8.72 10.8 12.76 NaOH Asp w/ HCHO 4.01 3.49 8.59 2.12 2.52 9.16 10.11 2.32 2.51 11.66 8.96 1.95 2.37 11.

Aspartic Acid: Acid VS Base Graph 14 12 10 8 6 4 2 0 0 2 4 5 6 8 10 12 14 15 16 18 20 HCl Asp w/o HCHO HCl Asp w/ HCHO NaOH Asp w/o HCHO NaOH Asp w/ HCHO .

53 1.42 NaOH Lys w/ HCHO 6.35 .74 1.81 1.16 7.58 11.25 7 5.55 1.65 1.67 1.61 1.Lycine: Lycine: Acid VS Base Data 0 2 4 5 6 8 10 12 14 15 16 18 20 HCl Lys w/o HCl Lys w/ HCHO HCHO 8.66 8.3 10.55 3 2.42 1.14 2.57 1.59 1.58 2.3 2.47 NaOH Lys w/o HCHO 9.79 6.36 9.52 1.03 11.94 2.04 1.38 7.36 6.04 9.5 2.87 1.47 1.54 10 10.76 7.32 1.

Aspartic Acid: Acid VS Base Graph 12 10 8 6 4 2 0 0 2 4 5 6 8 10 12 14 15 16 18 20 HCl Lys w/o HCHO HCl Lys w/ HCHO NaOH Lys w/o HCHO NaOH Lys w/ HCHO .

Questions .

Question y 1. What can account for Sorensen·s discovery that the endpoint of titration between an amino acid and a standard alkali (without formaldehyde) is not reached? .

Formaldehyde makes the solution more acidic when titrated with a standard base . and the proton titrated can be treated directly.Answer y Formaldehyde in excess readily combines with free unprotonated amino groups of amino acids to give dimethylol derivatives.

How do you account for this? .Question y 2. Compare the values obtained when the amino acid was titrated with HCl both in the absence and presence of formaldehyde.

This is due to the fact that formaldehyde ties down the amino groups. making the carboxyl hydrogen ion more available .Answer y In the presence of formaldehyde. titration curve was slightly lower.

Question y 3. At which pH will an amino acid exert its maximum buffering capacity? Why? Where in your graph is the buffering region for your amino acid? .

.Answer y An amino acid exerts its maximum buffering capacity at the plateau near or on its pKa values because it is in this area that there is equal concentration of proton donors and acceptors.

can you determine the nature of its R group i. From the titration curve of an amino acid.Question y 4. acidic or neutral? .e.. basic.

Answer y Yes. the nature of the R group can be determined from the titration of an amino acid through the determination of the pKa values for each dissociable group of an amino acid by extrapolating the midpoint of each buffering region or plateau within the curve. .

Answer y Since the ionic form of the amino acid present in an aqueous solution is dependent upon solutions pH. if: pH > pKa: basic (negatively charged) pH < pKa: acidic (positively charged) pH = pKa: zwitterion (no charge) isoelectric point .

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