CHEM 41.

22 - A Locker No: 16A

Lumba, Pamela Therese S. Date: 9/23/19
Extracting and Separating Organic Dyes
Introduction
With the rise of the global textile industry, the dye and dyeing industry grew to become an essential source
of economic growth for developing countries. For instance, the value of exports of dyes and dyestuffs
from India estimated $ 2.4 billion in 2017 (1). Biotechnology and contemporary trends also contributed
new techniques in the industry, such as “Biostoning,” which uses indigo dye and enzymes to provide
denim jeans their signature “stonewash look” (2). However, a waste product of the dyeing process is dye
effluent, which is difficult to treat and has led to adverse environmental and health impacts (1). To combat
these effects, developing countries turn to separation science, using techniques such as coagulation and
flocculation to treat dye effluent wastes (3). In addition, AirDye technology, which uses air instead of
water in textile dyeing, is quickly being adopted in developing countries to reduce dye effluent wastes in
the future of the industry (1).
Data and Results
Table 1. pH sensitivity summary table of Brilliant Blue FCF dye and Methyl Red by color change.
Dyes:
Brilliant Blue (Aqueous layer) Methyl Red (Organic layer)
Reagents that Color change pH sensitivity Color change pH sensitivity status:
manipulate pH observed from dye: status: observed from dye:
level:

Increase in pH level Light blue to bright Not sensitive to pH Orange to bright Sensitive to pH
by addition of green increase green increase
NaOH
Decrease in pH Bright green to Not sensitive to pH Bright green to Sensitive to pH
level by addition of light blue decrease orange decrease
HCL

Table 2. Polarity and solubility summary table of Indigo Dye and Methyl Red in different solvent systems.
Solubility of Dyes:
Solvents Polarity of Solvent: Indigo Dye Allura Red

Solvent A (water) Polar Insoluble Soluble

Solvent B (water- Polar > Nonpolar soluble Soluble > insoluble
acetone)
Solvent C (acetone- Nonpolar > Polar soluble Insoluble > soluble
hexane)
Solvent D (hexane) Nonpolar insoluble insoluble
 Polarity of Dyes:
Partially polar and nonpolar Polar

Figure 1. Distillation curve of Indigo dye extract in Temperature (℃) versus Time (minutes)
Discussion
Within the initial part of the experiment, various dyes were tested to investigate their sensitivity
to pH changes, change in color, solubility in certain solvents as well as deduce the identity of the unknown
dyes produced. From mixing distilled water and ethyl acetate with concentrated dye solution A, two dyes
were made, namely Brilliant Blue FCF and the conjugate base form of Methyl Red. The light blue aqueous
layer is Brilliant Blue FCF, given that the structure of the dye contains polar groups suitable for dissolving
in both water and ethyl acetate. The orange organic layer, however, is Methyl Red, given that the dye’s
structure lacks strong polar groups and has nonpolar benzene rings, rendering it almost insoluble with
water and ethyl acetate.
To test the dyes’ sensitivity to pH changes, NaOH and HCL were added to the mixture, this process
was summarized in Table 1. When NaOH was added, the two dyes changed color to a bright radioactive-
like green. This means that the dyes are sensitive to an increase in pH level. With the addition of HCL, the
bright green color reverted to the dyes’ original orange organic layer and blue aqueous layer. This change
in color also means that the dyes are sensitive to a decrease in pH level. Thus, both Brilliant Blue FCF and
Methyl Red are proven to be sensitive to pH changes given their change in color.
In terms of testing Indigo dye and Allura red’s solubilities, four different solvents were used with
differing polarities in the following order: polar (water), strongly polar and partially nonpolar (water-
acetone), strongly nonpolar and partially polar (acetone-hexane), and nonpolar (hexane). Upon adding
Indigo dye and Allura red into each solvent, it was concluded that Allura red is more soluble in polar
solvents, specifically water and water-acetone solvents, while Indigo dye is more soluble in partially polar
and nonpolar solvents like water-acetone and acetone-hexane solvents as shown in Table 2. The solubility
of the dyes is greatly affected by their structure and intermolecular actions with the solvent. For instance,
Allura red has numerous positively and negatively charged groups suitable for ion-dipole interactions with
polar substances such as water and acetone solvents. On the other hand, Indigo dye has large nonpolar
structures in the form of packed benzene rings with relatively small polar groups due to C=O and N-H
bonds which make it suitable for London dispersion and dipole-dipole interactions with substance that
are both partially polar and nonpolar like water-acetone and acetone-hexane solvents.
The experiment focused on the recovery of a solvent collected from the extraction of Indigo dye. In the
initial part of the experiment, 32 mL of Indigo dye extract was collected via liquid-liquid extraction, using
ethyl acetate as the solvent. The extract itself was colored light blue-purple and smelled like acetone or
nail polish (4).
The distillation of the extract can be graphically represented through a distillation curve, as shown
in Figure 1. Looking closer at the curve, the temperature gradually increased until the 6th minute, followed
by a significant increase in temperature from 30℃to 64℃. The reason behind this jump in temperature is
due to the extract’s phase transition from a liquid state to a solid state. As a result, 13 mL of the solvent
was recovered, leading to a %recovery of 40.625%. The solvent collected via distillation and the solvent
used to collect Indigo dye extract were thus identical since they both appeared in the form of a clear liquid
that smelled of acetone or nail polish. For greater a greater yield solvent, it is recommended that the
distillation process should exceed 10 minutes to allow for greater %recovery and more solvent distilled.
In conclusion, the experiment was successful in extracting solvent distilled from indigo dye extract
and the investigation of various dyes’ pH sensitivity polarity, solvent solubility and identities. The
researchers not only gain experience behind the principles of separation science but also a perspective on
separation science’s role in chemical cycling for environmental waste treatment.

Answers to POSTLAB Questions:

1. In part one of the experiment, the identity of the blue dye is Brilliant Blue FCF dye. It is not
possible that Indigo dye would be the identity of the blue dye since the structure of the dye
partially shows both polar and nonpolar characteristics. The structure of Brilliant Blue FCF
dye, contains several polar groups such as SO₃⁻, Na⁺ and N⁺ which makes the it a strong,
polar substance. Given that ethyl acetate is partially polar and nonpolar, and that water is
polar, both substances will freely interact with the dye’s polar groups, exchanging and
displaying intermolecular forces such as H-bonding, dipole-dipole forces and ion-dipole
forces, allowing the dye to dissolve. This agrees to the pattern that ‘like dissolves and, as a
result, the Brilliant Blue FCF becomes the aqueous layer.

2.
The most acidic proton in Methyl Red’s structure is the proton attached to oxygen, since it is the
most electronegative atom. Attracted to the electrons on the negatively charged oxygen of the
hydroxide ion, the proton detaches from Methyl Red and attaches to the Hydroxide ion,
facilitating an acid-base reaction.
3. The acidic form of Methyl Red carries the color of a dark, rich fuchsia. The benzene rings in
Methyl Red’s structure suggests that it is a nonpolar substance, thus it cannot fully dissolve
in ethyl acetate as it is a partially polar molecule. The conjugate base form of Methyl Red
carries a lighter shade of dark fuchsia as compared to its acidic form. Since the color is still a
vivid dark fuchsia, it can still be said that the conjugate base form of Methyl Red is also not
soluble in ethyl acetate as the benzene rings are still present in its structure. However, since
the conjugate base form of Methyl Red contains a polar group, specifically COO-, which has
some effect to the form’s solubility in ethyl acetate, it can be said that conjugate base form
of Methyl Red is relatively more soluble than Methyl Red’s acidic form. In general, however,
Methyl red in its conjugate base and acidic form is not soluble in partially polar ethyl acetate
since it is an overall nonpolar substance.
4. In solvent A, where water is the solvent, Allura Red dissolved while Indigo dye did not
dissolve. Looking at Allura Red’s structure, it is very polar due to the presence of O⁻ and Na⁺.
This means that Allura Red can form strong H-bonds, dipole-dipole bonds and ion-dipole
bonds with water since it has charged groups, following the pattern that ‘like dissolves like.’
Indigo dye, however, shows both polar and nonpolar characteristics. Because its structure
has a large nonpolar group in the form of benzene rings packed together, this makes the
dye strongly nonpolar, but with the presence C=O and N-H bonds, the dye is also partially
polar as well. As a result, because indigo dye is more nonpolar than polar, it doesn’t dissolve
completely in water. Thus, the vial is almost completely colored red.
In solvent B, composed of water and acetone, water is polar, but acetone has methyl
groups, which are nonpolar, and carbonyl groups, which are polar, making the substance
both polar and nonpolar (5). Indigo dye dissolves with the presence of acetone’s methyl
groups through the exchange of H-bonds from its small polar groups and London dispersion
forces with acetone’s carbonyl groups from its large nonpolar groups. Allura red, being
polar, dissolves in water and acetone through the presence of H-bonds, ion-dipole forces
and dipole-dipole forces due to dye’s charged regions. Thus, the vial is mostly colored red
with a small portion of blue on top.
In solvent C, composed of acetone and hexane, acetone is both polar and nonpolar
while hexane is strongly nonpolar due to its C-H bonds. Indigo dye again dissolves with the
interaction between its small polar groups and large nonpolar groups with acetone’s polar
methyl groups and nonpolar carbonyl groups through H-bonding and London dispersion
forces. Allura red, with its polar groups, dissolves in acetone since it is also polar through the
presence of H-bonds, ion-dipole forces and dipole-dipole forces but not with hexane as it is
a nonpolar substance. Thus, the vial was equally colored both red and blue with blue on the
top and red on the bottom.
In solvent D, composed of hexane, hexane is a strong nonpolar substance due to its C–H
bonds and symmetrical geometric structure (6). Both indigo dye and Allura Red were
insoluble in hexane as exhibited by the formation of red and blue droplets upon
 introduction of the dyes to the solvent. This is due to the polar groups present in both dyes
which makes it insoluble to the highly nonpolar hexane as a solvent.
5. From the experiment, it has been proven that Indigo Dye does not dissolve well in water
because of its large nonpolar benzene rings. Thus, the dye cannot work the same way other
water-based dyes do. Indigo white is needed since it is soluble in water, given to its polar O-
H groups which water can interact with. Once the fabric is soaked in Indigo white, the fabric
is exposed to air and indigo white will react with oxygen to form the indigo into the fabric.

6. The boiling point of the solvent can be deduced graphically as it is marked by the beginning
of plateaus in the distillation curve (7). In this case, the boiling point is 64℃, as shown in the
Figure above.
7. The solvent collected in the distillation process is akin to ethyl acetate with a boiling point of
77.1℃and a scent described to be close to acetone or nail polish. However, the experimental
results yielded a boiling point of 64℃. As a result, the %error is 16.9%. Two major reasons
behind these deviations to standard results may due to the inaccurate reading of the
thermometer since it was not in direct contact with the mixture and possible impurities
within in the extract.
Sample Calculations
% Recovered Solvent = (Recovered solvent volume in mL/Extract volume in mL) x 100
= (13mL/32mL) x 100 = 40.625%
%Error Boiling Point = (Expected Temperature - Observed Temperature/Expected Temperature) x 100 =
(77.1 ℃- 64℃/ 77.1 ℃) x 100 = 31.25%

References
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