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ROTATION OF SUCROSE

INTRODUCTION

It is necessary for sucrose (cane sugar) to be hydrolyzed (broken) into simpler sugars it can be
metabolized. In biological systems, the water reacts with sucrose to break it down with the help of the
catalyzing enzyme invertase, which can be found in human saliva. This experiment looks at the kinetics
of this same reaction which is slow as suggested by the need of an enzyme. Instead of the enzyme
catalyst, however, we will use hydrogen ions. We will examine different ways of extracting the rate
constant from experimental data assuming a first order reaction as we will be working under an excess of
water.

THEORY AND METHODS

The rate of reaction between sucrose and water catalyzed by hydrogen ion is followed by measuring the
angle of rotation of polarized light passing through the solution. The angle is measured with a
polarimeter. The reaction is

C12H22O11 C6H12O6 + C6H12O6 (1)

Sucrose Glucose + Fructose

Sucrose is dextrorotatory, but the resulting mixture of glucose and fructose is slightly levorotatory. This
is because the levorotatory fructose has a greater molar rotation than the dextrorotatory glucose. As the
sucrose is used up and the glucose-fructose mixture is formed, the angle of rotation to the right (as the
observer looks into the polarimeter tube) becomes less and less, and finally the light is rotated to the left.
The rotation is determined from the beginning ( o ) to the end of the reaction ( ). The algebraic

difference between these two readings is a measure of the original concentration of sucrose. It is
assumed that the reaction goes to completion so that practically no sucrose remains at "infinite" time. At
any time t, a number proportional to the concentration C of the remaining sucrose is obtained from the
difference between the final polarimeter reading and the reading of t at time t.
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The reaction proceeds too slowly to be measured in pure water. It is catalyzed by hydrogen ions. The
water is in such large excess that its concentration does not change appreciably and the reaction follows
the equation for a first-order reaction, even though two different kinds of molecules are involved in the
reaction. Thus the reaction-rate constant (k) may be calculated from the equation

1 o
k = ln (2)
t t

which is derived from:

C
ln = kt (3)
Co

where C is the concentration.

Method 1: As can be seen from equations (2) and (3), the rate constant can be determined if the
value is known. For reactions that go to completion within a few hours this idea is quite feasible but for
very slow reactions where might not be obtained, the following methods can be used:

Method 2: The so-called Guggenheim method avoids the infinity reading by taking data at equal time
intervals and splitting the measured data (rotation angles versus time) into two sets. Here is how it
works. For each measured data point, 2i at time t 2i (i=0,1,2, etc), another measurement is taken at
time t 2 ( i +1) = t 2i + t, where t is a fixed time taken as 10 minutes for our experiments. The second
measurement would correspond to 2 ( i +1) = 2i + 2 .

Now, if a plot of ln ( 2i - 2i + 2 ) versus time is prepared, the points will fall on a straight line of slope k!
If t is too small, there will be a large percentage error in 2i - 2i + 2 .

Note that the fancy subscripts are introduced simply to show the data split in abstract terms. Note that
your 5 minutes spaced actual experimental data collection sequence will not work as it is. Rather the data
has to be split into two sets; 2i as demonstrated above and 2i +1 (i=0,1,2,..) set to be treated in the same
way.

The equation for this method may be derived as follows using 2i set (the same applies to the other set
obviously!). Remember that the difference between polarimeter readings is proportional to the
concentration of sucrose remaining in the experiment. From the integrated form of the first-order reaction
differential equation,

C = Co e k t (4)

Using equation (2) to express the concentrations C2i and C2i + 2 and given that the concentrations differ in
time by the fixed interval t, we can show that:
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C2i C2i + 2 = Co e k t 2 i (1 e k t ) (5)

Taking logarithms, we have:

ln( C2i C2i +2 ) = kt2i + ln [Co (1 e k t )] i = 0,1, 2, .. (6)

Thus the slope of a plot of ln( C 2i C 2i + 2 ) versus time is -k. Because readings are proportional to
actual concentrations of sucrose remaining, the slope of ln( 2i 2i + 2 ) versus time will be the same.

Method 3: This method is based on a non-linear fit that does not necessitate the infinity reading in the
first method nor does it require Guggenheims clever data splitting at equal time intervals.

The following equation showing the non-linear dependence of rotation on time can be derived from eq (2)
above:

t = ( 0 ) e k t + (7)

A nonlinear fit could then be used to determine the following optimal parameters:

A = ( 0 ) , k and B = (8)

Based on your experimental data points (rotation angle as a function of time), use Solver in excel to
carry out the fit11. Estimate the error on fitted rate constant using the same type of spreadsheet11.

PROCEDURE:

Each group will determine the rate constants for the rotation of sugars by one concentration of HCl. At
the end of the experiment each group will share the results of different concentrations.

1) Zero the polarimeter using a water (solvent) blank

2) Dissolve 20 g sugar in 100 ml H2O. Filter if necessary to give a clear solution.

3) Mix 25 ml sugar solution with 25 ml of the distilled water. Fill the cell with the solution and
measure the rotation angle o .

4) Mix 25 ml sugar solution with 25 ml of the acid. At the moment of mixing start timing.

5) Rinse cell with the new solution and fill it.

6) Take first reading on polarimeter. This first reading should be taken at 5 minutes.

7) Take readings every 5 minutes for 90 minutes.

8) Leave the setting as is and take an reading two days later or follow instructions by TA
regarding this value.
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Some specific instructions regarding data handling and analysis:

Show tables of your data and calculated values as required by the lab report format. Show all graphs and
calculations used to obtain rate constants by using methods 1 and 2. Show your fit results for method 3
and the error estimate associated with it. List in a single table the rates of reaction with different acid
concentrations arrived at by all methods and compare them to each other and to experimental values.

QUESTIONS:

1) a) Why do you use a monochromatic light source?

b) Why was plane polarized light used?

2) The reaction as studied in acidic medium should more properly be written as:

Sucrose + water glucose + fructose

a) The above equation describes a second order reaction, yet the reaction is considered to be
first order. Explain.

b) Describe how HCl catalyzes this reaction.

c) How does the rate depend on the acid concentration?

3) What are the advantages and disadvantages of the Guggenheim method?

4) Which one of the three methods would you consider most accurate and why? Discuss how they
differ in terms of sources of errors.

5) How would temperature affect the rate of the reaction?

OPERATION OF THE FULL CIRCLE POLARIMETER

The polarimeter measures , the rotation of the plane of polarized light produced by a liquid. One usually
uses the light of the sodium D line (a doublet at 589.0 and 589.6 nm in the yellow region). Here are the
operating instructions.

1. The sodium lamp should be about 4" from the mirror. With the scale set at about 50, adjust the
mirror for maximum brightness.

2. Reset the scale and vernier to 0 and make sure the penumbra angle setting is centered: - +

3. Fill the tube, and ensure that no air bubbles are in the light path. Do not tighten the screw too
much: strain on the end window may cause a spurious optical rotation.
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4. Be sure that the end plates are clear and dry outside. Put the tube into the polarimeter with the
enlargement (which accommodates any air bubbles) uppermost. Close the cover, and focus the
eyepiece sharply on the divisional lines by moving it back and forth. The central area will have
the appearance of one of the parts of Fig. 2.

5. Rotate the dial by means of the lever until the three parts of the central area are equally bright.
(figure 2-ii) This phenomenon persists over a very small range of values bordered on each side
by the two oppositely illuminated non-uniform areas (Fig. 2-i and iii). Do not use the rather
broad region where the field of view is bright and uniformly illuminated.

6. Read the optical rotation, , directly in degrees from the scale. The vernier permits reading to
twentieths of a degree. Use the positive side of the vernier scale for reading positive rotations,
and vice versa. Fig. 3 illustrates use of the vernier for reading both negative (-0.25) and positive
(+2.65) rotations.

7. If an absolute rotation is desired, a zero reading must be taken. To do this, repeat steps 4 to 6 with
distilled water in the polarimeter tube. The absolute rotation of the sample is its reading minus
the zero reading.

Figure 1 Schematic Drawing of a Polarimeter.

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REFERENCES

The background necessary to understand the theory behind this experiment is taken from "Experimental
Physical Chemistry" by Daniels et al, 7th edition, page 149.

1. Guggenheim, E.A., Phil. Mag., (7)2: 538 (1926).

2. Amis, E.S. and Jaffe, G., J. Chem. Phys., 10: 598 (1942).

3. Amis, E.S. and Holmes, F.C., J. Am. Chem. Soc., 63: 2231 (1941).

4. Bronsted, J.N., and Wynne-Jones, W.F.K., Trans. Faraday Soc. 25: 59 (1929).

5. Ciapetta, F.G. and Kilpatrick, M., J. Am. Chem. Soc. 70: 639 (1948).

6. Pennycuick, S.W., J. Am. Chem. Soc., 48: 6 (1926).

7. Scatchard, G., J. Am. Chem. Soc., 48: 2259 (1926).

8. Rendina, "Experimental Methods in Modern Biochemistry", p. 146.

9. Edelmann and Chapman, "Basic Biochemistry", Heinmann, 1979.

10. Clark, J.M. and Switzer, R.L., Experimental Biochemistry, 2nd ed., p. 147.

11. Daniel C. Haris, J. Chem Edu, 75: 119 (1998).