NAME PERIOD DATE

EQUILIBRIUM OF SUGAR MUTAROTATION

Polarimeters are used to determine the optical angle of rotation of optically active solutions. Light is passed through
a polarizing lens and propagates in one defined plane through a transparent solution and through a second polarizer
to reach a detector. If the solution is not optically active, and the second polarizer is aligned, the light at the detector
will be about as strong as it was when it came through the first polarizing lens. If the solution contains an optically
active compound, the plane of oscillation rotates requiring the user to move the second polarizer to find the angle
where the maximum amount of light reaches the detector.
According to Biot’s law, the degree to which light is rotated (αobserved) depends on the specific rotation constant of
the solute [α] multiplied by the solution concentration (c) and path length of light traveling through the sample (ℓ).
Concentration is reported in grams per milliliter (g/mL), and the PASCO polarimeter uses a sample cell with a path
length equal to 1.0 decimeter (dm). The units for [α] are degrees·mL·dm‑1·g‑1, usually abbreviated as degrees.
Biot's law: αobs= [α] × c × ℓ
For mixtures, optical activity is additive. A mixture of optically active molecules will rotate light by the sum of the
observed optical activities of each molecule. The concentration of each solute can be determined if the mole fraction
is applied to each solute concentration. Mathematically, this is an extension of Biot’s law. For example, in a mixture
with two optically active solutes:
αobs-mixture = (αsolute1 × cmole fraction1 × ℓ) + (αsolute2 × cmole fraction2 × ℓ)
Suppose pure D‑glucose is dissolved in water. Even though solid D-glucose is in the alpha or α‑configuration, a
solution will form a mixture of α‑ and beta, β‑configured isomers with a linear intermediate molecule in a reaction
called mutarotation.

Figure 1. Mutarotation of α‑glucose when dissolved in solution

Figure 1 shows how the hydroxyl group on carbon‑1 is inverted in the two ring structures. The α‑ and β‑rings are
diastereomers because they are isomers that are not mirror images of one another, and they are anomers because
they are ring configurations that only differ by one chiral center. As shown in the figure, the specific rotations are
different but are non‑opposite. Both anomers are dextrorotatory because the rotation values are positive. During
mutarotation, the observed optical rotation of the mixture changes as the α‑ and β‑anomers change their
concentrations. The two anomers interconvert very slowly over many hours during mutarotation, and eventually the
solution reaches equilibrium. The observed rotation of the D-glucose solution at equilibrium can be used to calculate
the α‑anomer percentage using the mixture equation:
αobs-glucose = (x × [αα-anomer]) + ((1-x) × [αβ-anomer]), or, αobs-glucose = x[αα-anomer] + [αβ-anomer] − x[αβ-anomer]
Since path length equals 1.0 dm, it has been omitted from the calculation. The units for [α] are abbreviated to
degrees and x is equal to the α‑anomer concentration (c) as g/mL in the solution. The β‑anomer percentage makes up
the remainder of the solution. Since the concentration of each anomer is not initially known, the mole fraction
equation 1 = (x) + (1‑x) can be applied in place of the g/mL concentrations. The mole fraction x is assigned to one
anomer and the mole fraction (1‑x) is assigned to the other anomer. If x was assigned to the β‑anomer instead of the
α‑anomer, the equation above would need to be revised but the concentrations would work out the same. The
concentrations as g/mL can be converted to percentage concentration by multiplying each by 100.

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To determine whether the solution favors the α‑ or β‑anomer at equilibrium, consider the average observed optical
rotation you would see if the solution had an equal concentration of each anomer. The average of 112° and 19°
equals 65.5°. If the two anomers have an equal concentration, the solution would have an αobs equal to 65.5°. If the
solution has an αobs above 65.5°, the anomer with the greater specific rotation has a higher concentration. If the αobs
is below 65.5°, the anomer with the lower specific rotation is favored.
Throughout your chemistry course, you have probably noticed many reactions proceed in one direction to
completion. It is not unusual for some products to be able to run the reverse reaction to re-form the starting
reactants. Reaction rates are fastest when reactants are first introduced in their highest concentration. As the
products build up, the reverse reaction eventually attains a rate to equal the forward reaction. At this point, the
reaction is said to be at equilibrium. A ratio using the equilibrium concentrations describes how much product is
formed compared to the starting amount of reactant. The value of this ratio is known as the equilibrium constant,
Keq. The equilibrium expression follows the general formula of:
[products]
Keq =
[reactants]
As an example, for the reaction 1A + 2B ⇌ 3C + 4D, the double arrow indicates a reversible reaction. The
equilibrium expression, otherwise known as the rate equation, looks like this:

[C]3 + [D]4
Keq =
[A] + [B]2
Notice the power terms come from the coefficients of the balanced equation. The balanced equilibrium reaction for
the mutarotation of D-glucose can be written as: α‑D‑glucose ⇌ β‑D‑glucose. The equilibrium expression for the
mutarotation reaction is stoichiometrically just 1 reactant to 1 product. The rate equation for the mutarotation of
glucose at room temperature is:
[β-D-glucose]
Keq =
[α-D-glucose]
The literature value for this transition is reported as Keq = 1.8 at 20 °C. This K value indicates that when a
D‑glucose solution has reached equilibrium, the α‑ and β-anomers are forming and re-forming but the concentration
of β‑D‑glucose molecules will be 1.8 times greater than the concentration of α‑D‑glucose molecules. Several factors
can cause the equilibrium to shift, so it is not unusual to find different values for Keq for this specific reaction that is
dependent on experimental conditions. Your goal is to find the equilibrium constant based on the specific rotation of
a glucose mixture experimentally determined under the conditions present.

Objectives
• Use polarimetry to determine the concentration of two different anomers in a D-glucose solution.
• Calculate the equilibrium constant of a D-glucose solution at equilibrium.

Materials and Equipment

• Data collection system • D-glucose solutions (4%, 6%, & 8%), ~15 mL each
• Polarimeter with sample cell • Distilled water, ~15 mL
• Beakers (3), 50-mL or larger • Marking pen or grease pencil
• Beaker, 250-mL or larger • Rinse bottle filled with distilled water

Safety
Follow regular laboratory safety precautions.

Procedure
1. Make sure the polarimeter is turned off and the USB cable is unplugged. Align the notch on the analyzer wheel
with the orientation line printed on the polarimeter.
2. Turn on the polarimeter. Open SPARKvue and connect the polarimeter to your device.

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3. Open the Quick Start file called Intensity versus Angle.

Note: If the file is not available, create a graph display with Intensity on the y-axis and Angle(°) on the
x-axis.

4. Label three small beakers as follows: 4%; 6%; 8%. The fourth beaker is a waste beaker.
5. Collect about 15 mL of each glucose solution in the appropriate beaker.
6. Remove the cap from one end of a clean, dry sample cell and set it aside. The cap includes an outer ring, rubber
washer, and end glass as shown in Figure 2. Handle the end glass by its sides.

Figure 2. Sample cell diagram

7. Remove the other cap. Make sure the end glass, rubber washer, and outer ring are assembled in the same order
as shown in Figure 2, then hand-tighten the cap to secure it.
8. Fill the sample cell with distilled water and slide the end glass over the threaded opening as shown in Figure 2.
Place the rubber washer and outer ring over the end glass and hand-tighten the cap.
9. Dry the outside of the sample cell and the visible portion of the end glass on either end of the cell.
10. Tilt the cell to allow all bubbles to collect in the air trap. If the cell has bubbles that will not move, gently tap the
cell on top of a paper towel pad until the bubbles move.
11. Set the sample cell inside the polarimeter. Slide the cell all the way to the right. If the cell has a logo or the air
trap is marked, take note of its location to keep cell position consistent across multiple runs.
12. Start collecting data. Move the analyzer by rotating the wheel towards the back of the polarimeter from zero
degrees to 360°. The wheel notch will align with the orientation line when a full rotation is complete.

Note: If a subsequent run begins around 360° instead of 0°, move the wheel just past 360°, stop
collecting data, and start a new run at the new wheel position.

13. Stop collecting data.

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14. Locate the first complete peak closest to zero degrees and complete the
following to determine the angle where maximum intensity is reached:
a. Open the Graph Tools and toggle the Move Tool to the Select Tool.

b. Add a selection box to capture the top third of the peak with
symmetry as shown in Figure 3.
c. Open the Curve Fits menu and apply a Gaussian fit to the selected
data. The function parameters will appear.
Figure 3. Symmetrical peak selection
d. The value for 'c' indicates the angle of maximum intensity. Record
this value in Table 1.
15. Pour the contents of the cell into the waste beaker. Shake out as much liquid from the cell as possible.
16. Rinse the cell with a few milliliters of 4% glucose solution. Repeat steps 8-15 with 4% glucose instead of
distilled water, followed by the two remaining glucose solutions to complete Table 1.
17. Remove both caps. Rinse the sample cell and cap components with distilled water. Shake excess liquid from the
cell and cap components. Loosely replace the cap components on the cell.
18. Calculate the observed angle of rotation for each D-glucose solution (αobserved) by subtracting the angle at
maximum intensity for the reference substance (distilled water) from the solution angle at maximum intensity:
αobserved = solution angle – reference angle. Report answers in Table 1.
19. Calculate the average observed angle of rotation for a D-glucose solution at equilibrium under the conditions
present in the lab.

Data Collection
Table 1. Angle of maximum light intensity for D-glucose solutions and average αobs

Substance Angle at maximum intensity (°) Observed angle of rotation (αobs)

Distilled water (solvent) N/A

4% D-glucose

6% D-glucose

8% D-Glucose

Average

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Questions and Analysis

1. Based on the calculated average observed rotation for the D-glucose solution, does the equilibrium solution
favor the α‑anomer, β‑anomer, or neither? Explain your answer.

2. Use the mixture equation to calculate the α‑anomer concentration in g/mL; refer to Figure 1 for the specific
rotation of each anomer:
αobs-glucose = x[αα-anomer] + [αβ-anomer] − x[αβ-anomer]
Show your work in the space provided.

3. Convert the α‑anomer concentration from g/mL to percent by multiplying by 100, then determine the β‑anomer
concentration based on this percentage. What is the percentage concentration of each anomer?

4. Use the percentage concentrations to calculate the equilibrium constant for the D-glucose solution at equilibrium
under the conditions present in your lab classroom; show your work. The value for Keq at 20 °C equals 1.8.
Comment on your calculated Keq value; what does it say about the ratio of products compared to starting
reactants, and is it reasonable based on the given Keq value? Why or why not?

5. Near freezing, the observed angle of rotation of a D-glucose solution at equilibrium is close to 80°. Determine
which anomer is favored at this temperature, calculate the equilibrium constant for these conditions, and use it to
defend your answer.

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