1

Concentration vs. Optical Activity

Research Question
“To what extent does the increase in concentration of D – (+) – lactose from 4.0% to 7.0% (m/v) (in
intervals of 0.5%) affects the angle of rotation it causes when dissolved in distilled water, in turn affecting
its optical activity, measured using a polarimeter?”
Introduction
When Medicinal Chemistry was introduced in my IB Chemistry class during the Year 1 of DP, before it was
removed from the course with Paper – 3, we studied about Thalidomide tragedy1 as an example of how the
same chemical compound in different isomeric forms can go from being useful to tragically harmful. As
seen from the tragedy itself, one isomer of the drug thalidomide helped in treating nausea in pregnant
women, while the other caused severe defects in unborn children and death of fetuses. When I studied
further about optical isomers, I learned that they are two non – superimposable mirror images of the same
compound with equal number of atoms and bonds. Further, what made optical activity interesting was
during my IB Chemistry class, I also learned about how optical isomers can cause a rotation in plane
polarized light passed through. An anticlockwise rotation is caused by the (-) enantiomer whereas the (+)
enantiomer causes a clockwise rotation. Further, due to my inclination towards medicinal sciences, I
searched more about optical isomerism and found that various organic compounds like sugars (glucose,
fructose, lactose etc.) exist as enantiomers.
Researching through optical isomerism, I came across the technique polarimetry2 which is used to quantify
the angle of rotation caused in the plane polarized light by the optically active compound that it is passed
through. I also discovered several types of polarimeter like Biquartz, X – Ray, semi – automatic, full
automatic etc. with varying construction and designs. My school lab had the semi – automatic chemical
polarimeter which is primarily used for chemical solutions and has a rotator that is manually controlled, thus
I decided to base my investigation‟s methodology on it. Fascinated I researched further and came across a
paper, "Determination of Specific Rotation of Lactose”3 by Charles Francis B. Sanchez et. al., stating
how polarimetry can be used to determine the specific rotation caused by dextro – lactose, but this was done
only at a specific concentration. But on research, I found that dextro – lactose found in milk (human and
cow both) is present in a concentration ranging from 4.0% to 5.0% (m/v) in cow’s milk and upto 7.0% (m/v)
in human milk.4 This made me wonder how a change in concentration of a substance would affect the
optical activity of a substance because as concentration increases the number of particles in the solution
would increase, thus having more particles to deflect the light passing through, causing a change in the angle
of plane polarized light. Thus I chose to perform this investigation to determine the optical activity of dextro
– lactose dissolved in water at varying concentrations ranging from 4.0% to 7.0% (m/v) in an interval of
0.5%, leading to the following research question:

1
Kim, James H., and Anthony R. Scialli. "Thalidomide: The Tragedy Of Birth Defects And The Effective Treatment Of Disease". Toxicological Sciences, vol
122, no. 1, 2011, pp. 1-6. Oxford University Press (OUP), doi:10.1093/toxsci/kfr088. https://pubmed.ncbi.nlm.nih.gov/21507989/ Web. 16 Aug 2020.
2
"OPERATION AND TYPES OF POLARIMETERS". 2020, https://www.mrclab.com/operation_and_types_of_polarimeters. Web. 16 Aug 2020.
3
Sanchez, Charles Francis B. et. al. "Determination Of Specific Rotation Of Lactose | Chirality (Chemistry) | Chemistry." Scribd. May 1, 2018.
https://www.scribd.com/document/377899206/Determination-of-Specific-Rotation-of-Lactose Web. 18 Aug 2020.
4
"Lactose - An Overview | Sciencedirect Topics." Sciencedirect.com. Jul. 23, 2016. https://www.sciencedirect.com/topics/agricultural-and-biological-
sciences/lactose Web. 18 Aug 2020.
 2

“To what extent does the increase in concentration of D – (+) – lactose from 4.0% to 7.0% (m/v) (in
intervals of 0.5%) affects the angle of rotation it causes when dissolved in distilled water, in turn affecting
its optical activity, measured using a polarimeter?”
To answer the research question, I decided to prepare seven different solution of lactose in distilled water
with varying concentrations of 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5% and 7.0% (m/v) and measured the
deflection in the angle of plane polarized light caused by each using a polarimeter, to determine the effect of
concentration on the angle of rotation of plane polarized light.
Optical activity of organic compounds is an important property in determining which enantiomer is useful
and which is harmful for human body, like in the case of glucose, L – (+) – glucose is not processed by our
bodies because of its extreme chirality as compared to D – (+) – glucose.5 Another example is nicotine,
wherein (-) – nicotine is more toxic to human being than (+) – nicotine, and excessive intake of the former
can lead to vision loss or blindness, as stated by Stefanie B. Varghese et.al.6 in “The Effects of Nicotine on
the Human Electroretinogram.” Further, as lactose and other sugars are important filler in pharmaceuticals,
knowing about optical activity can be extremely helpful in determining their effectiveness as fillers or
catalysts in case of other substances. Thus, the study of optical activity is of utmost importance. This
research would also be helpful in finding the optimum concentration of lactose that causes the deflection
closest to its literature value7 of specific rotation i.e. +52.3°, making it worthy.
Background Information
Lactose is a common sugar found primarily in milk and its products. When dissolved in a solvent, lactose
exhibits two different isomeric forms – alpha (α) – lactose and beta (β) – lactose.8 Lactose is a disaccharide
that is made of ß-D-galactose and ß-D-glucose molecules bonded via a ß1-4 glycosidic linkage. Lactose has
the empirical formula C12H22O11 with a molecular weight of 342.3 g mol-1, with the following structure.

Figure 1 Lactose9
Lactose constitutes upto 7% solid content in milk, human and cow both. Further, lactose is the only known
source for galactose whereas glucose can be found in other food items as well like bread, rice, pasta,

5
"L-Glucose". Pubchem.Ncbi.Nlm.Nih.Gov, 2020, https://pubchem.ncbi.nlm.nih.gov/compound/L-
Glucose#:~:text=It%20is%20an%20enantiomer%20of%20an%20aldehydo%2DD%2Dglucose.&text=A%20primary%20source%20of%20energy,in%20fl
uid%20and%20nutrient%20replacement. Web. 18 Aug 2020.
6
Varghese, Stefanie B. et al. "The Effects Of Nicotine On The Human Electroretinogram". Investigative Opthalmology & Visual Science, vol 52, no. 13,
2011, p. 9445. Association For Research In Vision And Ophthalmology (ARVO), doi:10.1167/iovs.11-7874.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3293416/ Web. 18 Aug 2020.
7
“Determination of Lactose content in milk using polarimetry” Topac.com. Nov. 12, 2019.
http://www.topac.com/documents/Polarimeter%20lactose%20in%20milk.pdf Web. 18 Aug 2020.
8
Vinodhini, K. & Bharathi, R. & Srinivasan, K.. (2018). Studies on the concentration dependence of specific rotation of Alpha lactose monohydrate (α-
LM) aqueous solutions and growth of α-LM single crystals. IOP Conference Series: Materials Science and Engineering. 310. 012006. 10.1088/1757-
899X/310/1/012006.
https://www.researchgate.net/publication/323536957_Studies_on_the_concentration_dependence_of_specific_rotation_of_Alpha_lactose_monohydrate_a
-LM_aqueous_solutions_and_growth_of_a-LM_single_crystals Web. 18 Aug 2020.
9
"Lactose Molecule - Chemical And Physical Properties." Scienceofcooking.com. May 17, 2016. https://www.scienceofcooking.com/lactose.htm Web. 20
Aug 2020.
 3

vegetables, fruits etc. As galactose is a part of various macromolecules which make up the nerve cell
membrane in the human body, consumption of lactose rich products is required for humans. Further, lactose
is also associated with the adsorption of calcium, zinc etc in infants and is also essential for the growth of
bifid bacteria in the gut which plays an important role countering the age – related decline in the human
immune system.
Optical activity10 is a property of organic chiral molecules wherein the optical isomers are non –
superimposable mirror images of each other having equal number of atoms and bonds with a varying spatial
arrangement. Both the non – superimposable images are called enantiomers of each other. The physical
properties of enantiomers like boiling and freezing point, density etc. are same and the difference in
chemical properties is only seen in chiral environments. Further, when a plane polarized light is passed
through the enantiomers, each rotates the light at the same magnitude but in opposite directions. An
anticlockwise rotation is caused by the (-) enantiomer whereas the (+) enantiomer causes a clockwise
rotation.

Figure 2 Enantiomers of Glucose

Further, as lactose shows optical activity when subjected to plane polarized light and finds an important use
as pharmaceutical filler, studying its angle of rotation is important. Further, with increase in concentration,
the numbers of molecules present in the solution increases, there are more molecules to deflect the plane
polarized light and cause a higher deflection in the angle of rotation.
Polarimeter
Polarization is a property of waves that tells about the orientation of the oscillations taking place when the
wave travels. Further, a polarimeter can be used to detect the deflections taking place in these waves in the
form of plane polarized light, when passed through an optically active solution. This device can be used to
calculate the concentration of the solution through which the plane polarized light was passed and is
especially used for sugars, peptides etc. It consists of a plane polarized light source, an analyzer, a graduated
circle to measure the angle of rotation and cylindrical tubes to carry the sample solutions. The pointed light
source is passed through the sample solution and its deflections are noted by the analyzer which produces a
wave graph on the Logger Pro or Lab quest it is connected to, using which the angle of rotation can be
noted.

10
"Optical Isomers". Chem.Purdue.Edu, 2020, https://www.chem.purdue.edu/jmol/cchem/opti.html. Web. 20 Aug 2020.
 4

Figure 3 Working of a Polarimeter11

Hypothesis
As stated in the Background information, with an increase in the concentration of lactose from 4.0% to 7.0%
(m/v) in the solution, the number of lactose molecules available to deflect the plane polarized light would
increase, thus the extent of deflection caused was expected to be higher, thus the angle of rotation was
expected to be lower. It was also hypothesized that as the solution were made in distilled water, with
increasing concentration of lactose in it, the angle of rotation would move farther from the angle of
rotation of distilled water and closer to the literature value of the angle of rotation of lactose i.e. +52.3°.
Null Hypothesis
The increase in concentration of lactose in the solution would have no effect on the angle of rotation.
Variables
Independent Variables
Concentration of lactose was varied in distilled water from 4.0% to 7.0% (m/v) with an interval of 0.5% to
check the effect of changing concentration on the angle of rotation of plane polarized light. This range was
chosen as the concentration of lactose in cow milk is 4.0% - 5.0% (m/v) and goes upto 7.0% (m/v) in human
milk, thus acting as the independent variable in this investigation.
Dependent Variables
Angle of rotation of the plane polarized light would directly be affecting by the number of molecules
available to deflect the light, thus was dependent on the concentration of lactose and acted as the dependent
variable in this investigation.
Controlled Variables
Several other variables were controlled in this investigation to ensure uniformity in the method and
comparability in the results obtained. The volume of solution used in each trial was controlled at 10 cm3, by
filling the required volume in the polarimeter tube. This value was found to give the most measurable results
during the pre – lab trial. The speed and direction of rotation was controlled manually by completing one
rotation in 10 seconds, in clockwise motion. This was also determine during the pre – lab, but as it was to
be done manually, controlling it was a little bit difficult. The intensity of the light source was kept constant

11
"Understanding Polarimetry - Vernier." Vernier. Mar. 7, 2012. https://www.vernier.com/experiment/chem-o-6_understanding-polarimetry/ Web. 20
Aug 2020.
 5

throughout the research to ensure uniformity, by using the same light source in the polarimeter for all trials
that was set at 589 nm, as per the manual. The room temperature and pressure were tried to maintain by
performing the trials on consecutive days to ensure similar temperature and pressure.
Materials Required
Table 1 Apparatus required
S.No. Apparatus Specifications Quantity
1. Polarimeter ±0.1° 1
3 3
2. 10 cm Measuring Cylinder ±0.05 cm 1
3. Digital Weighing Balance ±0.001 g 1
4. Watch Glass - 1
5. Glass Rod - 1
6. Spatula - 1
7. Lab Quest - 1
8. Laptop with Logger Pro - 1
Table 2 Chemicals Required
S. No. Chemical Formula Manufacturer Quantity
1. β - D – Lactose C12H22O11 Sigma - Aldrich 12.000 g
2. Distilled Water H2O Prepared in Lab As Required

Figure 4 Materials Required

Safety Precautions, Environmental & Ethical Concerns
Lactose12 is not particularly harmful, but must not be inhaled and tasted. As it is a lab grade chemical, so to
avoid any accidental touch, I made sure to use safety gloves and lab coat during the experimentation.
Further, only the required amount of solutions were prepared to ensure no wastage of lactose, as it was an
expensive chemical that was imported from USA. The left over solutions were diluted with water before
disposing off in the sink. The solutions were not disposed near any plants or water bodies as it could be
harmful to the organisms in soil and water bodies.

12
“Lactose” Fishersci.com. Jun. 7, 2010. https://www.fishersci.com/store/msds?partNumber=AAH54447A1&productDescription=BETA-D-
LACTOSE+CONTS%3D+BETA+1KG&vendorId=VN00024248&countryCode=US&language=en Web. 21 Sept. 2020.
 6

Research Methodology
Preparation of Solutions along with Percentage uncertainties
To prepare 4.0% (m/v) lactose solution, I dissolved 0.400 g lactose in 10.00 cm3 water as per the following
calculations:
0.400 𝑔
𝑥 100% = 4.0% (𝑚 𝑣)
10.00 𝑐𝑚3
Similarly, for 4.5% (m/v) solution, I dissolved 0.450 g lactose in 10.00 cm3, 0.500 g for 5.0% (m/v) solution
and so on, as per the following table 3. Percentage uncertainties were calculated as follows, example
shown for 4.0% (m/v):
%∆ 𝑖𝑛 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑖𝑡𝑜𝑛 = %∆ 𝑖𝑛 𝑚𝑎𝑠𝑠 𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑 + %∆ 𝑖𝑛 𝑣𝑜𝑙𝑢𝑚𝑒 𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑
𝐿𝑒𝑎𝑠𝑡 𝑐𝑜𝑢𝑛𝑡 𝑜𝑓 𝑤𝑒𝑖𝑔𝑕𝑖𝑛𝑔 𝑏𝑎𝑙𝑎𝑛𝑐𝑒 0.001
%∆ 𝑖𝑛 𝑚𝑎𝑠𝑠 𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑 = ∗ 100.000% = ∗ 100.000% = 0.250%
𝑀𝑒𝑎𝑠𝑢𝑟𝑒𝑑 𝑉𝑎𝑙𝑢𝑒 0.400
𝐿𝑒𝑎𝑠𝑡 𝑐𝑜𝑢𝑛𝑡 𝑜𝑓 10 𝑐𝑚3 𝑚𝑒𝑎𝑠𝑢𝑟𝑖𝑛𝑔 𝑐𝑦𝑙𝑖𝑛𝑑𝑒𝑟 0.05
%∆ 𝑖𝑛 𝑣𝑜𝑙𝑢𝑚𝑒 𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑 = ∗ 100.000% = ∗ 100.000% = 0.5%
𝑀𝑒𝑎𝑠𝑢𝑟𝑒𝑑 𝑉𝑎𝑙𝑢𝑒 10.0
Table 3 Preparation of Solutions
Volume of
Concentration of Mass of
S. Water added %∆ in %∆ in %∆ in
Lactose solution Lactose added 3
No. (cm ± 0.05 mass volume concentration
(% m/v) (g ± 0.001 g) 3
cm )
1. 0.0 0.000 10.00 0.000 0.5 0.500
2. 4.0 0.400 10.00 0.250 0.5 0.750
3. 4.5 0.450 10.00 0.222 0.5 0.722
4. 5.0 0.500 10.00 0.200 0.5 0.700
5. 5.5 0.550 10.00 0.182 0.5 0.682
6. 6.0 0.600 10.00 0.167 0.5 0.667
7. 6.5 0.650 10.00 0.154 0.5 0.654
8. 7.0 0.700 10.00 0.143 0.5 0.643
Calibration Curve for Polarimeter
The polarimeter was connected to a lab quest and a laptop. To calibrate the polarimeter, I filled 10 cm3
distilled water in the polarimeter tube and started the data collection in Logger Pro. I then rotated the
analyzer slowly in clockwise direction until a full wave was formed on the Logger Pro graph. This graph
was saved as the calibration curve.
Procedure
A new file was opened on the Logger Pro connected with the polarimeter. I refilled the polarimeter tube
with 10 cm3 fresh distilled water and measured the angle of rotation of pure distilled water; by taking the
Gaussian fit of the first peak and the coefficient „A‟ gave the angle of rotation, as shown in Graph 1 below.
After that, I prepared 4.0% (m/v) lactose solution and poured it in the polarimeter tube and measured the
angle of rotation by rotating the analyzer slowly in clockwise direction. The same procedure was repeated
with all the lactose solutions from 4.5% to 7.0% (m/v) concentrations. The entire process was repeated three
times to obtain three reading for angle of rotation.
 7

Figure 5 Experimental Set - Up

Data Collection & Analysis
Qualitative Data
Lactose was a white solid that formed colorless and odorless solution on dissolving with water. As there was
only solvation taking place, not much physical change in the solution was observed.
Raw Data

Graph 1 Sample Curve for Distilled Water – Trial 1

Table 4 Angle of Rotation – Trials
Angle of rotation (° ± 0.1°)
S. No. Concentration of Lactose Solution (% m/v)
Trial 1 Trial 2 Trial 3
1. 0.0 96.93 95.79 96.80
2. 4.0 81.00 80.50 80.70
3. 4.5 77.20 77.30 77.60
4. 5.0 70.10 70.60 70.30
5. 5.5 66.80 66.20 66.40
6. 6.0 59.70 59.60 59.70
7. 6.5 55.40 55.70 55.50
8. 7.0 53.30 53.00 53.70
 8

Data Processing & Uncertainty Analysis

The trials were averaged and their uncertainty and standard deviation were calculated as follows:
𝑆𝑢𝑚 𝑜𝑓 𝑎𝑙𝑙 𝑡𝑟𝑖𝑎𝑙𝑠
𝐴𝑣𝑒𝑟𝑎𝑔𝑒 𝑎𝑛𝑔𝑙𝑒 𝑜𝑓 𝑟𝑜𝑡𝑎𝑡𝑖𝑜𝑛 =
3
𝑀𝑎𝑥𝑖𝑚𝑢𝑚 𝑎𝑛𝑔𝑙𝑒 − 𝑚𝑖𝑛𝑖𝑚𝑢𝑚 𝑎𝑛𝑔𝑙𝑒
𝑈𝑛𝑐𝑒𝑟𝑎𝑡𝑖𝑛𝑡𝑦 𝑖𝑛 𝑎𝑛𝑔𝑙𝑒 𝑜𝑓 𝑟𝑜𝑡𝑎𝑡𝑖𝑜𝑛 =
2

(𝑥 − 𝑚𝑒𝑎𝑛)2
𝑆𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝐷𝑒𝑣𝑖𝑎𝑡𝑖𝑜𝑛 =
𝑁
Where x = angle of rotation in a given trial
Mean = average of all trials
N = number of trials
Sample Calculations
For 4.0% Lactose solution:
81.00 + 80.50 + 80.70
𝐴𝑣𝑒𝑟𝑎𝑔𝑒 𝑎𝑛𝑔𝑙𝑒 𝑜𝑓 𝑟𝑜𝑡𝑎𝑡𝑖𝑜𝑛 = = 80.73°
3
81.00 − 80.50
𝑈𝑛𝑐𝑒𝑟𝑎𝑡𝑖𝑛𝑡𝑦 𝑖𝑛 𝑎𝑛𝑔𝑙𝑒 𝑜𝑓 𝑟𝑜𝑡𝑎𝑡𝑖𝑜𝑛 = = ±0.25°
2

(81.00 − 80.73)2 + (80.50 − 80.73)2 + (80.70 − 80.73)2

𝑆𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝐷𝑒𝑎𝑣𝑖𝑡𝑎𝑖𝑜𝑛 = = 0.25
3
Similarly, the data was processed for all the lactose solutions and is shown below.
Table 5 Angle of Rotation, Uncertainties and Standard Deviation
S. Concentration of Lactose Average Angle of Uncertainty in Angle of Standard
No. Solution (% m/v) rotation (° ± 0.1°) Rotation (±°) Deviation
1. 0.0 96.51 0.57 0.62
2. 4.0 80.73 0.25 0.25
3. 4.5 77.37 0.20 0.21
4. 5.0 70.33 0.25 0.25
5. 5.5 66.47 0.30 0.31
6. 6.0 59.67 0.05 0.06
7. 6.5 55.53 0.15 0.15
8. 7.0 53.33 0.35 0.35
Data Analysis
The aim of the investigation was to determine the effect of increasing concentration of lactose in distilled
water on the deflection of plane polarized light caused by it, measured in terms of the angle of rotation using
a polarimeter. As the data collected and processed in Table 5 shows that the angle of rotation decreases
from 80.73° to 53.33°, as the concentration of lactose solution increases from 4.0% to 7.0% (m/v). This
happens because with an increase in the concentration, the number of lactose molecules in the solution
increases and thus there are more number of molecules present to cause a greater extent of deflection in the
path of plane polarized light, causing the final angle of rotation to be lower. This trend can also be seen in
the graph 2 below.
 9

Graph 2 Concentration of Lactose vs. Optical Activity

Another observation made from the collected data was that as the concentration of lactose increased and
reached closer to 7.0%, the angle of rotation caused by the solution became closer to the literature value of
angle of rotation of lactose i.e. +52.3°. Thus, it can be inferred that the technique of polarimetry can be
used to determine the purity of optically active compounds.13
Evaluation
The aim of the research to was to observe the effect of increasing concentration of lactose in distilled water
on the angle of rotation shown by the solution in the presence of plane polarized light and the observed
results followed the expected trend and showed increasing deflection in plane polarized light with lower
angles of rotation, as the concentration increased. The value of angle of rotation kept moving closer to the
literature value of +52.3° for β – D – lactose, thus showing that the trend and results obtained were accurate,
thus strengthening the results and methodology of the investigation. To further, ensure the accuracy and
consistency of the results, the uncertainty and standard deviation were calculated which came out in the
ranges of 0.0° to 0.4° (Table 5) and 0.1 to 0.4 (Table 5), respectively. As both these ranges were minute, the
results were accepted as accurate and consistent, adding to the strength of the investigation.
Further, the percentage deflection in angles of rotation, from the angle of rotation of distilled water i.e.
90.0°, was also calculated for each concentration of lactose solution to determine the extent of deflection
caused in the path of plane polarized light. This was done as follows:
𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝐷𝑒𝑓𝑙𝑒𝑐𝑡𝑖𝑜𝑛
𝐴𝑛𝑔𝑙𝑒 𝑜𝑓 𝑅𝑜𝑡𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝐷𝑖𝑠𝑡𝑖𝑙𝑙𝑒𝑑 𝑊𝑎𝑡𝑒𝑟 − 𝐴𝑛𝑔𝑙𝑒 𝑜𝑓 𝑟𝑜𝑡𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑠𝑜𝑙𝑢𝑡𝑖𝑜𝑛
= ∗ 100%
𝐴𝑛𝑔𝑙𝑒 𝑜𝑓 𝑅𝑜𝑡𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝐷𝑖𝑠𝑡𝑖𝑙𝑙𝑒𝑑 𝑊𝑎𝑡𝑒𝑟

13
Pharmacopoeia, Lactose. "Lactose Purity According To Pharmacopoeia :: Anton-Paar.Com". Anton Paar, 2020, https://www.anton-paar.com/in-
en/products/applications/lactose-purity-according-to-
pharmacopoeia/#:~:text=Lactose%20monohydrate%20in%20compliance%20with%20EU%2FUS%20pharmacopoeias&text=The%20US%20and%20Eur
opean%20pharmacopoeias,specific%20rotation%20of%20lactose%20monohydrate.&text=The%20specific%20rotation%20has%20to,cell%20length%20
of%201%20dm. Web. 16 Nov. 2020.
 10

For 4.0% (m/v) lactose solution:

96.51 − 80.73
𝑃𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝐷𝑒𝑓𝑙𝑒𝑐𝑡𝑖𝑜𝑛 = ∗ 100% = 16.34%
96.51
Similarly, the remaining percentage deflections were calculated and are tabulated below.
Table 6 Percentage Deflection
S. No. Concentration of Lactose Solution (% m/v) Percentage Deflection (%)
1. 4.0 16.34
2. 4.5 19.83
3. 5.0 27.12
4. 5.5 31.13
5. 6.0 38.17
6. 6.5 42.46
7. 7.0 44.74
As seen from the table above, the extent of deflection in plane polarized light increases from 16.34% to
44.74% as the concentration of lactose in distilled water increases from 4.0% to 7.0% (m/v), which also
aligns with the expected trend and further strengthens the accuracy of the results obtained. A weakness of
the methodology used could be that the polarimeter had to be controlled manually, which could have led to
human errors in speed and degree of rotation, adding some discrepancies in the data. These could be reduced
by using fully automatic polarimeters.
Conclusion
The investigation carried out to determine the extent to which the increase in concentration of dextro –
lactose (4.0% to 7.0% (m/v) in intervals of 0.5%) affects the angle of rotation it causes when dissolved in
distilled water, in turn affecting its optical activity, measured using a polarimeter, showed a clear trend
that aligned with the hypothesis made. The optical activity of the solution increased with increase in the
concentration from 4.0% to 7.0% (m/v), as the number of molecules in the solution increased and thus
causing a greater extent of deflection in the plane polarized light and hence a lower angle of rotation value
was shown. The angle of rotation decreased from 96.51° in pure distilled water to 80.73° in 4.0% (m/v)
lactose solution to 53.33° in 7.0% (m/v) lactose solution, as seen in Table 5 and Graph 2. Also, as the
concentration increased from 4.0% to 7.0% (m/v), the percentage deflection in the plane polarized light, as
compared to that caused by distilled water increased from 16.34% to 44.74%, respectively, as seen from
Table 6. This could be because as the concentration increases, there are more number of particles per unit
volume of the solution, thus more number of molecules are encountered by the plane polarized light causing
a greater deflection in the light and making the angle of rotation smaller. Further, another observation was
that at the highest concentration of 7.0% lactose, there was maximum deflection in the plane polarized light
and the angle of rotation was closest to the literature value of angle of rotation for lactose i.e. +52.3°. As the
observed trends and results coincided with the hypothesis made, it was accepted and the null hypothesis
became invalid.
 11

Limitations & Improvements

The most prominent limitation of this investigation was using a polarimeter with a manual analyzer, which
had to be rotated by me. Though, I tried to maintain a uniform speed of rotation for every trial, but certain
human errors might have caused some deviations and errors in the collected data for angle of rotation. This
could either be improved by using a time based approach for the investigation i.e. by fixing the time of data
collection instead of rotating the analyzer manually and then noting the angle, or by using a fully automatic
and digital polarimeter to ensure precision and accuracy in the data collected.
Another limitation of the investigation was that I used only seven different concentrations of lactose solution
and could not determine the exact concentration which shows the literature value of angle of rotation i.e.
+52.3°. This could be improved by taking a bigger range of concentrations and determining the exact
concentration that shows the angle of rotation as 52.3°.
Further, some minor errors in measurement of mass of lactose powder and volume of solutions could have
caused discrepancies in the collected data. These can be minimized by further increasing the number of
trials.
Future Extensions
The investigation can further be extended to determine the concentration of lactose that shows the angle of
52.3°. Moreover, the investigation could be repeated with other sugars like glucose and galactose to check
the effect of their changing concentration of the angle of rotation. The change in angle of rotation of lactose
solution, when broken down into glucose and galactose in the presence of different catalysts like sulfuric
acid etc. can also be determined and compared. Further, the impact of change in temperature of a sugar
solution on its angle of rotation and also be investigated. The inversion of sucrose along with it rate kinetics
could also be a part of further investigations.
 12

Bibliography
Journals
 Sanchez, Charles Francis B. et. al. "Determination Of Specific Rotation Of Lactose | Chirality
(Chemistry) | Chemistry." Scribd. May 1, 2018.
https://www.scribd.com/document/377899206/Determination-of-Specific-Rotation-of-Lactose Web.
18 Apr. 2020.
 Vinodhini, K. & Bharathi, R. & Srinivasan, K.. (2018). Studies on the concentration dependence of
specific rotation of Alpha lactose monohydrate (α-LM) aqueous solutions and growth of α-LM single
crystals. IOP Conference Series: Materials Science and Engineering. 310. 012006. 10.1088/1757-
899X/310/1/012006.
https://www.researchgate.net/publication/323536957_Studies_on_the_concentration_dependence_of
_specific_rotation_of_Alpha_lactose_monohydrate_a-LM_aqueous_solutions_and_growth_of_a-
LM_single_crystals Web. 18 Apr. 2020.
Web Sources
 “Determination of Lactose content in milk using polarimetry” Topac.com. Nov. 12, 2019.
http://www.topac.com/documents/Polarimeter%20lactose%20in%20milk.pdf Web. 18 Apr. 2020.
 "Lactose - An Overview | Sciencedirect Topics." Sciencedirect.com. Jul. 23, 2016.
https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/lactose Web. 18 Apr.
2020.
 "Lactose Molecule - Chemical And Physical Properties." Scienceofcooking.com. May 17, 2016.
https://www.scienceofcooking.com/lactose.htm Web. 20 Apr. 2020.
 "Understanding Polarimetry - Vernier." Vernier. Mar. 7, 2012.
https://www.vernier.com/experiment/chem-o-6_understanding-polarimetry/ Web. 20 Apr. 2020.
Safety Data Sheets
 “Lactose” Fishersci.com. Jun. 7, 2010.
https://www.fishersci.com/store/msds?partNumber=AAH54447A1&productDescription=BETA-D-
LACTOSE+CONTS%3D+BETA+1KG&vendorId=VN00024248&countryCode=US&language=en
Web. 21 Apr. 2020.