CH116-1L | Physical Chemistry Laboratory

1st Semester SY 2019-2020

Turbidity, Refractive Index and Ultraviolet-Visible Spectrophotometry

(Experiment 3)
Group 5
Quizon, Cedric Phillip1; Read, Jezreel; Rivera, Xenn Mar1; Santos, Steffany1; Sion, Joshell Mharielle1; Tacis, Kate Marie1,
Almendrala, Michelle2

1Students,CM116-1L/B2, School of Chemical, Biological, Materials Engineering and Sciences, Mapúa University; 2 Professor, School of Chemical, Biological, Materials
Engineering and Sciences, Mapúa University;

ABSTRACT

This study show three prevalent methodology in determining properties of fluids. Fluids can contain suspended solid matter
consisting of particles of many different sizes. While some suspended material will be large enough and heavy enough to settle
rapidly to the bottom of the container if a liquid sample is left to stand (the settable solids), very small particles will settle only
very slowly or not at all if the sample is regularly agitated or the particles are colloidal. These small solid particles cause the
liquid to appear turbid. Also, the refractive index shows the ratio of the velocity of light in a vacuum to its velocity in a specified
medium. Furthermore, UV-ViS uses light in the visible and adjacent ranges. The absorption or reflectance in the visible range
directly affects the perceived color of the chemicals involved. Each experiments were carried out in accordance to their
respective procedural standards. The researchers, upon discussion, were able to conclude specific findings and learning upon
conducting the experiment.

Keywords: UV-Vis, Turbidity, Refractive Index, Spectrophotometry, fluid property

INTRODUCTION

The experiment are divided into three major parts: index. For a solution of sugar, the refractive index can be
Refractometry (refractive index of sugar solution); used to determine the sugar content.
Turbidimetry (water analysis); and Spectrophotometry (UV-
Vis). The principles behind each part are discussed as The samples involved in the experiment are Coca Cola,
follows: Orange juice, Royal, Sprite, and 2.0 M sugar. In such fluids,
the refractive index is the measure of dissolved or
REFRACTOMETRY submicronic particles. The Brix scale relates the refractive
index to sugar concentration. This shows how the sugar
The refractive index of a transparent optical medium, also concentration can be controlled and monitored throughout
called the index of refraction, is the factor by which the phase the process, by continuously measuring the refractive index.
velocity is decreased, in relation to the velocity of light in
vacuum, when the optical medium has interacted with the TURBIDIMETRY
electromagnetic radiation. For transparent materials, the
refractive index is a dimensionless quantity higher than 1. Turbidity is defined as the optical property that causes light
to be scattered and absorbed rather than transmitted in
Since the refractive index is a fundamental physical property straight lines through a sample. As light passes through pure
of a substance, it is often used to identify a particular water, it travels along a relatively undisturbed path. The light
substance, confirm its purity, or measure its concentration. passed through fluids with suspended particles present, is
Refractive index is used to measure solids, liquids, and greatly distorted by absorption or scattering, caused by these
gases. Most commonly it is used to measure the particles.
concentration of a solute in an aqueous solution. A
refractometer is an instrument used to measure refractive

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 CH116-1L | Physical Chemistry Laboratory
1st Semester SY 2019-2020

Moreover, turbidity is considered as a good measure of the To determine the turbidity of different water sample
quality of water. Several water samples such as distilled using the turbidity meter.
water and tap water from Mapua University, Nature Spring
water, Sip, and 0.1 M Ca(OH)2 are measured for their To determine the refractive index of different juice
turbidity values using specialized optical equipment in the samples using the refractometer.
laboratory. A light is directed through a water sample, and
the amount of light scattered is measured. The unit of
measurement is called Nephelometric Turbidity Unit (NTU),
which comes in several variations. Turbidity is affected by MATERIALS AND METHODS
several factors in water: presence of dissolved and
TURBIDITY
suspended solids, size and shape of particles and
composition of the particles. The more total suspended
The students prepared four aliquot of water samples to be
solids in the water, the murkier it seems and the higher the used for the experiment. The water samples used were:
turbidity. Low turbidity values indicate high water clarity; high Cardinal water; Mapua tap water; Sip distilled water; and
values indicate low water clarity. Nature Spring drinking water. Each sample were put in a
bottle inserted in the turbidimeter. The data obtained from the
SPECTROPHOTOMETRY (UV-Vis) testing were recorded.

Every chemical compound absorbs, transmits, or reflects REFRACTIVE INDEX

light (electromagnetic radiation) over a certain range of
wavelength. Spectrophotometry is a method to measure how The students used the Refractor-meter to get all the required
much a chemical substance absorbs light by measuring the data. Using a dropper, students dropped little amount of
sample in the refractor-meter. The small amount of sample
intensity of light as a beam of light passes through sample
was placed inside the measuring prism. Moreover, the
solution. The basic principle is that each compound absorbs refractor-meter was then moved into the light coming from
or transmits light over a certain range of wavelength. the window to view the data inside.

Several samples of Cobalt Nitrate with different

concentrations and one unknown are prepared to be tested. ULTRAVIOLET-VISIBLE SPECTROPHOTOMETRY
The specific spectrometry apparatus utilized in the The students have prepared seven (7) duplicates of 10
experiment is the UV-visible spectrophotometer, where it milliliter sample while the lab assistant prepared an unknown
uses light over the ultraviolet range (185 - 400 nm) and solution in the Uv-vis spectrophotometer. Using the equation
visible range (400 - 700 nm) of electromagnetic radiation (M1V1=M2V2), the students were able to get the volume of
spectrum. With the spectrophotometer, the amount of a the concentrated solution, where M1 is the molarity of
known chemical substance (concentrations) can also be concentrated solution, V1 is the volume of concentrated
determined by measuring the intensity of light detected. solution, M2 is the molarity of dilute solution, V2 is the volume
of dilute solution. Cobalt Nitrate (Co(No3)2) was the
concentrated solution used for this experiment. After
computing the volume of concentrated solution, the students
started using a pipette to measure different amounts of
Objectives concentrated solution then it was diluted in water. After
dilution, the students transferred the diluted solution in the
The objectives of the experiment are the following: cuvette and was placed in the UV-VIS spectrophotometer
and then recorded the data that was shown in the computer.
To determine the concentration of an unknown
sample using the ultraviolet visible spectrophotometer. RESULTS AND DISCUSSION

Turbidity

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 CH116-1L | Physical Chemistry Laboratory
1st Semester SY 2019-2020

Procedural turbidity testing of the water samples and the 2.0 M Sugar 51.3 51.0 51.8 51.4
calcium hydroxide using the turbidimeter showed the
following results in Table . The values obtained shown in the Table 2 shows the refractive index of different sugar-
data uses Nephelometric Turbidity Units (NTU). Water containing compound. The data shows that the 2.0 M sugar
samples number 1, 3, and 4 showed that the turbidity sample has the highest refractive index compared to the
other samples. The reason behind this is that the sugar
measure is at 0.00 NTU, this explains that there is no
concentration is directly proportional to the refractive index
suspended solid matter in the water from these samples. which means that the higher the sugar concentration, the
Moreover, the results shown by testing Sample 2 and 5 more the light bends which results to a higher refractive
showed turbidity values of 2.16 NTU and 1.23 NTU, index. This is because, the higher the sugar concentration,
respectively. These shows that there is a presence of the thicker the solution will be, creating a denser medium
suspended solid matters within the sample aliquot. resulting to a higher refractive index. The data shows the
carbonated drinks and orange juice have lower refractive
Table 1. Turbidity units measured using a turbidimeter index and most are of almost the same value. This indicates
Sample Brand/Type Turbidity, NTU that these drinks have lower sugar concentration than of the
1 Cardinal Water 0.00 2.0 M sugar.
2 Mapúa tap water 2.16
3 Nature Spring 0.00
4 Sip 0.00 CONCLUSIONS AND RECOMMENDATIONS
5 0.1 M Ca(OH)2 1.23
The first part of the experiment is measuring the turbidity,
Furthermore, the propensity of particles to scatter a light wherein the researchers used five (5) different samples
beam focused on them is now considered a more meaningful namely, distilled water, tap water, mineral water (nature
measure of turbidity in water. Turbidity measured this way spring), purified water (sip), and 0.1 M Calcium hydroxide.
uses an instrument called a nephelometer with the detector Turbidity is a measure of how clear the sample is, in
set up to the side of the light beam. More light reaches the reference to the presence of bacteria and total suspended
detector if there are lots of small particles scattering the solids. Based on the experiment, distilled, mineral, and
source beam than if there are few. The units of turbidity from purified water have 0.0 NTU while tap water and 0.1 M
a calibrated nephelometer are called Nephelometric Ca(OH)2 have 2.16 NTU and 1.23 NTU, respectively.
Turbidity Units (NTU). To some extent, how much light Therefore, those two samples that have a high level of
reflects for a given amount of particulates is dependent upon turbidity have a lot of suspended particles and light cannot
properties of the particles like their shape, color, and pass through. As for the remaining three samples that have
reflectivity. For this reason, and the reason that heavier 0.0 NTU, they have a low turbidity since there are no particles
particles settle quickly and do not contribute to a turbidity present and are clearer, this also means that they are safe
reading, a correlation between turbidity and total suspended to drink. Thus, measuring the turbidity of water can assess
solids (TSS) is somewhat unusual for each location or the appearance and acceptability of drinking water supplied
situation. to consumers as well as the effectiveness of household
water treatment and safe storage.

For the second part of the experiment, refractive index of

different beverages were measured. Refractive index is used
to measure the concentration of a solute in aqueous solution
and in this experiment the sugar content of the samples were
determined. The refractive index of samples with low
concentration is much greater than that of the sample,
Refractive Index creating a large refraction angle and a low reading. While
those with high concentrations, the refractive index are lower
Table 2. Refractive index in three trials
than the sample, thus, having a small angle and a high
Sample Trial 1 Trial 2 Trial 3 Average
reading. Therefore, a low reading means that the beverage
Coca-cola 7.0 7.4 7.5 7.3
Orange Juice 5.1 6.0 5.8 5.6 have low sugar content and a high reading have a high sugar
Royal 5.0 5.5 5.0 5.2 content. This method can help determine beverages with
Sprite 5.4 5.0 5.3 5.2

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 CH116-1L | Physical Chemistry Laboratory
1st Semester SY 2019-2020

high sugar content, which can increase the risk of dangerous Table 3. Encountered problems with corresponding solutions
conditions such as heart attack and blood sugar problems. Encountered Problems Proposed Solutions
Always keep in mind proper
Not sanitized hands where the
sanitation before starting a
Lastly, the ultraviolet-visible spectroscopy was used to blood sample was collected
laboratory experiment.
determine the analyte concentration or the chemical Practice precision and accuracy
Refraction index problems
conversion of a component in a solution, where it should also in doing experiments
satisfy the Beer-Lambert’s Law that states that the
absorbance of a solution is directly proportional to the
concentration of the absorbing species in the solution. Based
on this experiment, the Beer Lambert’s Law was satisfied in
a way that as the volume of the sample increases, the
absorbance increases as well, thus making it directly
proportional with each other. Having a high absorbance
means that most of the light are absorbed by the sample and
only a small amount of the light is detected by the detector,
the opposite goes for those with low absorbance.

REFERENCES

Haswell, S. (1991). Atomic Absorption Spectrometry;

Theory, Design and Applications. Elsevier, Amsterdam.

McGaRefractive Index and Application (2014). Retrieved

from: https://www. Omics online.org/refractive-index-and-
its-applications-2157 7544.1000e117.php?aid=14768

Turbidity: Description, Impact on Water Quality, Sources,

Measures (2008). Retrieved from:
https://www.pca.state.mn.us/sites/default/files/wq-iw3-
21.pdf

Spectrophotometry (2019.) Retrieved

from:https://chem.libretexts.org/ Bookshelves/ Physical
_and
_Theoretical_Chemistry_Textbook_Maps/Supplemental_M
odules_(Physical_and_Theoretical_

Chemistry)/Kinetics/Reaction_Rates/Experimental_Determi
nation_of_Kinetcs/Spectrophotometryrthy, G. (2018, March

Walsh, Alan - Biographical entry. Retrieved from

Encyclopedia of Australian Science:
http://www.eoas.info/biogs/P000860b.htm
Nabil, R.B. (2010). Sample preparation for flame atomic
absorption spectroscopy: an overview. Rasayan J Chem.
2010;4:49–55.

APPENDICES

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