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## U0900023J U084997N Practical 1: pH & Buffers

Introduction
Objective:
To understand buffers, their buffering regions and the Henderson-Hasselbalch equation.

Background:
Buffers work by resisting changes in pH. Only weak acids and bases, which can form conjugate bases and acids respectively can be buffers. A pH meter is used to measure the H+ concentration in any given solution, the result is temperature dependent, as such, the machine must be calibrated at a known pH and temperature.

## Materials & Methods:

Various buffers, namely, Histidine Monohydrochloride, Potassium Phosphate and Tris-HCl, are used in 3 separate experiments. Titration was used not only to determine buffering capacity but also Ka values and the effect of temperature.

3.1.4

## Data handling and questions

(a) Number of moles of Histidine present in solution: 9.73 x 10-4 mol (b) (c) (d) (e) See graph below pKa values: 5.94 & 9.94 It shows maximal buffering capacity at pH 6. The effective buffering range is pH 6 to 9. Using method (i) will give a more accurate estimate of pKa, since it is assumed the NaOH has not been prepared accurately, then any calculations based on initial known values would be off. However, in the case of the graph, it is merely a rough estimate, and hence any discrepancies will not be significant enough to give false readings. (i) for 5ml of NaOH: pH is 6.20, as compared to 6.14 in the experiment. (ii) for 12ml of NaOH: pH is 8.90 as compared to 8.73 in the experiment. Differences in calculated pH values and experimental pH values differ. This might be due to experimental errors and errors in substance preparation. (i) Number of ionisable groups in Histidine at initial pH: 2 (ii) Groups responsible for observed pKas: NH3+ and =NH+

(f)

(g)

## (h) Structures of ionic species in histidine that participate in buffering:

Results
Part 1:
pH 4.82 5.17 5.37 5.53 5.68 5.78 5.86 5.94 6.04 6.14 6.22 6.33 6.42 6.54 6.65 6.75 6.95 7.17 7.33 7.66 Vol of NaOH /ml 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 mol NaOH 0.000025 0.00005 0.000075 0.0001 0.000125 0.00015 0.000175 0.0002 0.000225 0.00025 0.000275 0.0003 0.000325 0.00035 0.000375 0.0004 0.000425 0.00045 0.000475 0.0005 mol NaOH/mol Hist 0.025693731 0.051387461 0.077081192 0.102774923 0.128468654 0.154162384 0.179856115 0.205549846 0.231243577 0.256937307 0.282631038 0.308324769 0.334018499 0.35971223 0.385405961 0.411099692 0.436793422 0.462487153 0.488180884 0.513874615

8.09 8.39 8.57 8.73 8.84 8.97 9.07 9.17 9.26 9.35 9.43 9.53 9.61 9.71 9.81 9.94 10.07 10.22 10.41 10.65 10.88 11.07 11.21 11.34 11.42 11.48 11.5

10.5 11 11.5 12 12.5 13 13.5 14 14.5 15 15.5 16 16.5 17 17.5 18 18.5 19 19.5 20 20.5 21 21.5 22 22.5 23 23.5

0.000525 0.00055 0.000575 0.0006 0.000625 0.00065 0.000675 0.0007 0.000725 0.00075 0.000775 0.0008 0.000825 0.00085 0.000875 0.0009 0.000925 0.00095 0.000975 0.001 0.001025 0.00105 0.001075 0.0011 0.001125 0.00115 0.001175

0.539568345 0.565262076 0.590955807 0.616649538 0.642343268 0.668036999 0.69373073 0.71942446 0.745118191 0.770811922 0.796505653 0.822199383 0.847893114 0.873586845 0.899280576 0.924974306 0.950668037 0.976361768 1.002055498 1.027749229 1.05344296 1.079136691 1.104830421 1.130524152 1.156217883 1.181911614 1.207605344

Q 3.2.4 Part 1 Tris-Hcl buffer seems to be better at buffering when an alkaline solution is added and Potassium Phosphate buffer seems to be better when an acidic solution is added. Part 2 Effective buffering range for Potassium Phosphate buffer is pKa 1 that is pH range 5.8 - 7.8. Effective buffering range for Tris-Hcl buffer is from pH 7.1 9.1. So for the phosphate which functions optimally at pH7.2 Tris-Hcl buffer should be used. Although it is not better than the potassium phosphate buffer in the case if large quantities of alkali is added its effective buffering range shows that is will still be a buffer is an acid added. In the case of the potassium phosphate buffer it is not a good offer if the solution starts to become acidic. Q 3.3.4 Temperature of Solution(oC) 25 oC 4 oC pH of Tris-Hcl Buffer pH of Potassium Phosphate Buffer 7.11 7.13

1.47 1.87

From these results it can be concluded that lowering the temperature of the solutions causes the pH of the respective solutions to rise. Tris-HCl buffer and potassium phosphate buffer both contain weak acids. HA H+ + ADissociation is a bond breaking process thus it requires energy in order to proceed. Thus the forward reaction is endothermic and the reverse reaction is therefore exothermic. When the buffer solutions were cooled to 40C, the buffer tried to remove the disturbance characterized

as the loss of heat energy by shifting the equilibrium towards producing more undissociated acid. This is to counter the energy lost as heat so as to restore the equilibrium in a way. With decreased dissociation, [H+] in solution would be lower accounting for an increase in pH.