STEWART WANI R114504F

BIOCHEMISTRY EXPERIMENT 1

In order to carry out various metabolic and biomechanical functions in order to maintain homeostasis, cells need enzymes to enhance the reactions. These enzymes each act best at a particular pH. Some processes cause either an increase or a decrease in pH, causing the environment of the cells to digress from optimum conditions, and compromising the enzyme activity. Because of this, the body contains several systems in place in order to regulate pH. AIM: To determine the concentrations of sodium hydroxide and sodium carbonate in an unknown mixture of both in solution MATERIALS: 1) Alkali solution (containing roughly 3g/liter NaOH and 1g/liter Na2CO3) 2) 1M HCl stock solution 3) Methyl orange indicator (pH range 3.2 4.4), 0.05g dissolved in 50ml water 4) Phenolphthalein (pH range 8.3 10.0), 0.5g in 50ml ethanol and 50ml water METHOD: Make 100ml of 0.1M HCl from stock. Place 25ml aliquot of unknown alkali solution (containing (NaOH AND Na2CO3) into a conical flask Titration: Phase 1: Add 2 drops of phenolphthalein indicator into the solution in the Flask. Titrate to end point with 0.1M HCl. Note volume (V1) Phase 2: Add 2 drops methyl orange indicator to the above solution and continue titration to the new end point. Note new volume (V2). Above procedure to be done in triplicate PROCEDURE 1st Procedure 2nd Procedure 3rd Procedure Mean TITRE Final Initial V1(ml) Final Initial V2(ml) Vol.(ml) Vol.(ml) Vol.(ml) Vol.(ml) 11.2 0.3 10.9 14.7 11.2 3.50 24.3 10.4 10.17ml 14.7 0.4 9.60 10.00 28.1 14.0 3.63ml 24.3 10.4 3.80 3.60

CALCULATIONS: Ai) 2HCl + NaOH + Na2CO3 NaHCO3 + 2NaCl + H2O Reacting ratio: HCl: NaOH: Na2CO3 2 1 1 Volume of HCl used = 0.5 x 10.17ml = 5.085ml 2HCl + NaOH + Na2CO3 NaHCO3 + 2NaCl + H2O NaHCO3 + HCl NaCl +CO2+H2O Volume of HCl used = (0.5 x 10.17ml) + 3.63ml = 8.715ml B) Concentration of NaOH = 0.1 x 5.085 25 = 0.02034mol/L = 0.02034 x 40 = 0.8136g/L ii) Concentration of Na2CO3 = 0.1 x 8.715 25 = 0.03486mol/L = 0.03486 x 106 = 3.695g/L DISCUSSION: In Phase 1, the mean value of V1 was slightly lower than expected, due to stopping the titration before completely reaching the end-point. As a result, V2 in Phase 2 is slightly higher than expected, in order to compensate for the slightly low value of V1. In Phase 1, the HCl reacts with both NaOH and Na2CO3 in the conical flask. Since it would take 1 mole of HCl to neutralize 1 mole of NaOH, and another mole of HCl to react with 1 mole of Na2CO3 in Phase 1, half of the titrated volume V1 would neutralize NaOH while the other half would react with Na2CO3. In Phase 2, all of the NaOH has been neutralized, and the HCl reacts with NaHCO3 which was formed in the previous reaction. Since 1 mole of HCl reacts with 1 mole of NaHCO3 to form NaCl, CO2 and H2O, the entire titrated volume V2 would react with NaHCO3 to complete the neutralization of Na2CO3. Together with Phase 1, it would take half of V1 plus V2 to completely neutralize Na2CO3. The number of moles of HCl which neutralized NaOH can be determined by multiplying its molarity by the volume that it took to neutralize NaOH (0.5 x V1). This would be equal to the number of moles of NaOH, because the 1 mole of HCl neutralizes 1 mole of NaOH. The concentration of NaOH in moles/liter can then be determined by dividing by the volume of the unknown solution, and converted to grams/liter by multiplying that concentration by the gram-molecular mass of NaOH.

The number of moles of HCl which neutralized Na2CO3 can be determined by multiplying its molarity by the volume that it took to neutralize Na2CO3. This would be equal to twice the number of moles of Na2CO3, because 2 moles of HCl would be needed to neutralize 1 mole of Na2CO3: 1 mole in Phase 1 and another in Phase 2. The concentration in moles/liter is then obtained by dividing by the volume of the unknown solution, and converted to grams/liter by multiplying that concentration by the grammolecular mass of Na2CO3. QUESTIONS: Ai) an acid is a proton-donor ii) A base is a proton-acceptor Bi) pH = -log[H+]. Lehninger (2005) says, The greater the acidity of a solution, the lower its pH. ii) pH = -log(6.5x10-5) = 4.19

Ci) a buffer is a solution which is able to resist a change in pH due to an addition of acid or base. It consists of a weak acid and its conjugate base. ii) K2 K1 CO2 + H2O H2CO3 H+ + HCO3-

K1 = [H+][HCO3-] H2CO3 K2 = [H2CO3] [CO2][H2O] K1 = [H+][HCO3-] K2[CO2][H2O] K1K2[H2O] = K3 = [H+][CO3-] [CO2] An increase in acidity or alkalinity would result in a change in every single component in the solution, either to neutralize excessive acid, or to produce acid in order to counter a rise in pH. Whenever H+ or OH- is added to a buffer, the result is a small change in the ratio of the relative concentrations of the weak acid and its anion and thus a small change in pH. The decrease in concentration of one component of the system is balanced exactly by an increase in the other. The sum of the buffer components does not change, only their ratio. (Lehninger (2005))

D) The end-point is not easily discernible when using an indicator, and there is a high risk of titrating past it, compromising the accuracy of results. CONCLUSION: Buffer systems are essential for the proper maintenance of bodily function. Without them, there would be no way for enzymes to establish their optimum working environment, which would result in poor health, and possibly even death.

REFERENCES: Thomas M. Devlin, Biochemistry with Clinical Correlations, Wiley-Liss, NewJersey 2006 David L. Nelson, Michael M. Cox, Lehninger Principles of Biochemistry 4th Edition, Freeman and Co., 2005 Jeremy M. Berg, John L. Tymoczko, Biochemistry 5th Edition, Freeman and Co., 2002 P.C. Champe, R.A. Harvey, D.R. Ferrier, Biochemistry: Lippincotts Illustrated Reviews, Lippincott, Williams and Wilkins 2005