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In the Classroom

edited by

Ed Vitz
Kutztown University
Kutztown, PA 19530

Developing an Invisible Message about Relative


Acidities of Alcohols in the Natural Products Henna,
Turmeric, Rose Petals, and Vitamin A
Brahmadeo Dewprashad* and Latifa Hadir
Department of Science, Borough of Manhattan Community College, City University of New York,
New York 10007
*bdewprashad@bmcc.cuny.edu

Introducing chemical concepts to students by using concrete examples and interest-arousing demonstrations is a wellaccepted pedagogy. Demonstrations and activities pertaining
to invisible inks are popular (1). Natural indicators obtained
from common fruits, vegetables, teas, and flowers have been used
in a number of instructor demonstrations and simple student
experiments (2-9). These activities are engaging and attract
students' attention (1-9). We have developed a demonstration
utilizing invisible inks and solutions of three naturally occurring
dyes as developing agents. The demonstration not only attracts
students' attention but also illustrates the concepts of relative
acidities of alcohols, resonance stabilization, and keto-enol
tautomerization.
A core concept in undergraduate organic chemistry is that
the acidity of a functional group increases if the anion formed
(from loss of H) is stabilized. Both resonance and inductive
effects are considered when evaluating the relative acidities of
different molecules. However, many students rely more on recall
of memorized facts rather than on deductive reasoning when
predicting relative acidities. This may be because many students
have difficulties writing resonance structures. These students
may feel that they can arrive at a correct answer to predict
relative acidities (or relative reactivities) without drawing resonance structures. The demonstration can be used to introduce
exercises that provide students with additional practice drawing
resonance structures and predicting relative acidities.
The demonstration uses dilute basic solutions as invisible
inks. The inks are made visible using naturally occurring dyes
that undergo pH-dependent structural and color changes.
One of the developing agents is an ethanolic solution of the
2-hydroxy-1,4-naphthoquinone, which gives henna, a natural dye
used in hair colorants and in body decorations, its characteristic
color. Resonance theory predicts that 2-hydroxy-1,4-naphthoquinone should lose the hydrogen atom of its vinyl alcohol under
mild basic conditions as the corresponding anion formed is
resonance stabilized (Scheme 1). Delocalization of the electrons
(of the anion) results in a shift of the wavelengths absorbed by the
compound and a resultant change in color. A solution of
2-hydroxy-1,4-naphthoquinone is yellow under acidic conditions
but changes to reddish orange under basic conditions. A message
written on a filter paper with a basic solution such as dilute
Na2CO3 or NaOH becomes invisible on evaporation of the
solvent. When a solution of 2-hydroxy-1,4-naphthoquinone is
sprayed onto the filter paper, the deposited base reacts with the
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dye and the anion formed is reddish orange, displaying the


invisible message. If a dilute HCl solution is sprayed onto the
displayed message, it becomes blurred and disappears due to
reformation of 2-hydroxy-1,4-naphthoquinone. The displayed
message and its disappearance are shown in Figure 1. An
alternative is to use an ethanolic extract of henna as a developing
solution. The message developed has a greenish-orange tint. The
green color probably arises from the chlorophyll extracted along
with the 2-hydroxy-1,4-naphthoquinone from the henna, which
is made by pulverizing leaves from the plant Lawsonia inermis.
Another developing agent is an ethanolic extract of turmeric, a spice commonly used in Asian and African food. The
extract is yellow owing to the presence of curcumin (Figure 2),
the compound that gives curry its characteristic yellow color and
reputed health benefits. Curcumin exists predominantly as the
enol and is yellow under acidic conditions but changes to a red
color under basic conditions (10). Delocalization of the electrons
of the anion (shown in the supporting information) results in a

Figure 1. Appearance (left and center) and disappearance (right) of


messages on filter papers sprayed with a solution of 2-hydroxy-1,4naphthoquinone. (The identifying cuts are not shown.)
Scheme 1. Reaction of 2-Hydroxy-1,4-naphthoquinone with a Base and
the Resultant Resonance-Stabilized Product

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10.1021/ed800014k Published on Web 12/18/2009

In the Classroom

Scheme 2. Predominant Reaction of Cyanidin Diglucoside with a Weak


Base and the Resultant Resonance-Stabilized Product

Figure 2. Stuctures of curcumin and vitamin A.

shift in wavelength absorbed, which changes the color of the


compound. On spraying with dilute HCl solution the message
disappears.
A natural extension is to discuss the structural changes in
cyanidin, a pigment molecule responsible for the red, blue, and
purple color of many plants. Another developing agent used is an
ethanolic extract of rose petals. The compound that gives roses
their red color is cyanidin diglucoside. This compound (which
has a cyanidin moiety substituted with two glucose units) is red,
but under mildly basic conditions (pH 8-9) it loses an H to
give, predominantly, the anion that has a blue color (11, 12). The
reaction is shown in Scheme 2. Spraying with a dilute HCl
solution makes the message disappear. Providing students with
the structure of cyanidin diglucoside and asking them to predict what changes will occur under mild basic conditions
requires them to consider which hydrogen atom would be lost
most easily and the relative contributions of the various resonance structures. This example illustrates a resonance structure
in which a charge is neutralized. The exercise also provides
students with practice drawing more challenging resonance
structures.
An ethanolic solution of retinol, vitamin A (Figure 2), is
used as a standard yellow dye solution that does not change color
under basic conditions. Vitamin A is also yellow but is much less
acidic than 2-hydroxy-1,4-napthoquinone as the anion formed
(when it loses an H) is not resonance stabilized. The structure
of vitamin A (given in the supporting material) indicates that its
acidity is likely to be similar to that of ethanol. Vitamin A is not
likely to react with Na2CO3 and NaOH but only with strong
bases such as NaH. However, there is no change in color, even
when the anion is formed; the electrons on the oxygen are not in
conjugation with the double bonds and cannot be delocalized.
Vitamin A is not effective in making visible, a message written
with a Na2CO3 or NaOH solution.
Materials
0.1 M Na2CO3 (0.1 g in 10 mL of distilled H2O)
0.1 M NaOH (0.04 g in 10 mL of distilled H2O)
0.1 M HCl (10 mL)
0.040 M (0.50 g in 72 mL ethanol) 2-hydroxy-1,4-naphthoquinone
0.040 M (0.50 g in 44 mL ethanol) vitamin A, store in a
refrigerator
4 g of henna
2 g of ground turmeric
3 red roses

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4 Whatman #4 24.0 cm diameter filter papers


2 cotton swabs
4 spray bottles
disposable gloves
scissors
4 tea bags

Preparation of Extract from Henna


Empty the tea from two tea bags. Place 4.0 g of henna into
one bag, fold at the open end, and staple. Place the bag in the
remaining empty tea bag, fold, and staple. Place 50 mL of ethanol
in a 100 mL beaker, cover, and heat to just below the boiling
point. Turn off the heat. Immerse the henna in the hot ethanol.
Cover the beaker and wait 30 min. Filter the liquid and place it in
the spray bottle.
Preparation of Extract from Turmeric
Empty the tea from two tea bags. Place 2.0 g of ground
turmeric into one bag, fold at the open end, and staple. Place the
bag in the remaining empty tea bag, fold, and staple. Place 50 mL
of ethanol in a 100 mL beaker, cover, and heat to just below the
boiling point. Turn off the heat. Immerse the turmeric in the hot
ethanol. Cover the beaker and wait for 30 min. Filter the liquid
and place it in a spray bottle.
Preparation of Extract from Rose Petals
Remove the petals from three roses. Place 75 mL of ethanol
in a 200 mL beaker, cover, and heat to just below the boiling
point. Turn off the heat. Immerse the rose petals in the hot
ethanol. Cover the beaker and wait for 30 min. Filter the liquid
and place it in a spray bottle.

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In the Classroom

Preparation of Filter Paper with Invisible Message


Dip a cotton swab in a 0.1 M solution of NaOH and use it
to write the word NaOH on each of a pair of Whatman #4
filter papers. Use a scissors to make a cut of 1 cm at the edge
of each of the filter papers with NaOH. The cut is used to
identify the paper. Repeat the procedure on four additional
filter papers using a fresh cotton swab and 0.1 M Na2CO3 to
write Na2CO3. Make two 1 cm cuts at the edges of each
of these papers as a form of identification. Allow the filter
papers to dry until the marks become invisible. This preparation of the filter paper should be done at least 15 min before
the demonstration to facilitate evaporation of the aqueous
solvent.
Hazards
Wear safety goggles and gloves when preparing the dye
extracts and dye solutions, writing the messages, and spraying the
developing solutions. These steps should be done in a fume hood.
In addition, spray the filter papers with developing solutions very
lightly and point away from yourself and students. The HCl and
NaOH solutions are corrosive. Also, both 2-hydroxy-1,4naphthoquinone and vitamin A are irritants and can be harmful
if they come in contact with the skin, are inhaled, or are
swallowed. Ethanol is flammable and should be kept away from
heat sources. In addition, the ethanolic extracts and solution of
dye should be properly disposed of in designated organic waste
bottles.
Presentation and Discussion
At the beginning of class, hold up the filter paper and invite
students to look for any written messages. Students are usually
puzzled and pay rapt attention at this stage. Invite one of the
students to look closely for any written words and to share his or
her observation with the class. Select the filter paper with the
single cut and spray with the solution of 2-hydroxy-1,4-naphthoquinone. Students are usually surprised when they notice that
from the blank paper, the word NaOH shows up in bright
orange. Provide the students with the structure of 2-hydroxy-1,4naphthoquinone and ask them to explain the reason for the
appearance of the message. Students usually are not able to
explain the color change. This is a good opportunity to introduce
the concept of relative acidity of alcohols. Point out that ethanol,
C2H5OH has a pKa of around 16; it does not react with solutions
of NaOH because the resulting conjugate acid would be H2O,
which has a pKa of 15.73 (13). Remind students that an acid
reacts with a base to form a weaker conjugate acid. Discuss the
stabilization (of the alkoxide anion) by inductive effects using as
an example CF3CH2OH, which has a pKa of 12.43, and explain
how the net electron-withdrawing effects of the three fluoride
atoms results in increased acidity (13). Point out that these
alcohols do not react with weak bases and react only to a limited
extent with the stronger metal hydroxide bases. Discuss the effect
of resonance stabilization of the alkoxide anion using as an
example, phenol, C6H5OH, pKa = 9.87 (13). The fact that
phenol is acidic enough to react with NaOH is rationalized by
the delocalization of the anion of the phenoxide. Have students
draw the different resonance structures for the phenoxide ion.
Next, use p-nitrophenol, p-NO2C6H5OH, (pKa = 7.15) and
p-aminophenol, p-NH2C6H5OH (pKa = 10.46), as examples to
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show where both resonance and inductive effects can combine to


affect the acidity of alcohols (13). The electron-withdrawing
-NO2 group increases the acidity of the phenolic -OH while
the electron-donating -NH2 group decreases it. Revisit the
reason for the message displayed by 2-hydroxy-1,4-naphthoquinone (Scheme 1). Ask students to draw the structure of the anion
formed and its resonance structures. At this stage, they are usually
able to draw the structure and can also explain the color change
observed.
Hold up one of the filter papers with the two cuts to the
class. Ask students whether the message (written with Na2CO3)
would also become visible when sprayed with the solution of
2-hydroxy-1,4-naphthoquinone and the reason(s) for their answers. Many students are likely to indicate that the message will
not be visible because Na2CO3 is not a strong enough base. They
are usually under the impression that a -COOH functionality
but not an -OH reacts with Na2CO3. Spray the paper with
the dye solution; the message becomes visible (Figure 1). Point
out that the results are consistent with the reported pKa values
of 4.70 for 2-hydroxy-1,4-naphthoquinone and 6.37 and 10.31,
respectively, for H2CO3 and HCO3-, the conjugate acids
of CO32- (14, 15). Alternatively, the message can be displayed
by spraying with an ethanolic extract of henna. The message
is displayed but has a greenish-orange color. If this developing solution is used, inform students that henna is crushed
leaves and the green tinge is likely due to the presence of
chlorophyll extracted along with the 2-hydroxy-1, 4-naphthoquinone.
Ask students to predict what would happen to the message if
the filter papers are now sprayed with 0.1 M HCl. Many students
are likely to indicate that the message disappears due to reformation of 2-hydroxy-1,4-naphthoquinone. Hold up one of the
papers with the visible message and spray with the dilute HCl.
The message becomes blurred and indecipherable and then
disappears. Students are usually pleased that their prediction is
validated.
Provide the students with the structure of vitamin A and ask
whether it can be used to develop a message written with
Na2CO3. Without closely examining the structure of vitamin
A, a number of students are likely to say that the message will
show up. Spray the vitamin A solution onto the filter paper
marked with Na2CO3. The message does not show up. Ask
students whether the message written with NaOH will become
visible if sprayed in a similar manner. Some of the students will
likely point out that the anion formed is not resonance stabilized
and, as such, the molecule is not acidic enough to react with
NaOH. Spray the filter paper marked with the NaOH solution;
no message appears. Explain that NaOH does not react with
vitamin A, but that if a stronger base such as NaH is used, a
reaction would occur as vitamin A likely has a pKa near to that of
ethanol. Point out that the loss of H from vitamin A is not
expected to lead to a color change as the anion formed is not in
conjugation with the double bonds and the charge is not
delocalized.
Continue by providing students with the structure of
curcumin and explain that it exists predominantly as the enol
form (12). Ask whether an invisible message written with
Na2CO3 will show up if sprayed with a curcumin-containing
extract from turmeric. Many students are likely to indicate that it
would. Ask them to explain the reason for their answers. In
discussing their responses, have them identify the most acidic

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In the Classroom

hydrogen atom and write resonance structures for the anion


formed from loss of that hydrogen. Ask students to compare the
acidity the phenolic -OHs in curcumin with that of phenol. As
the ortho -OCH3 group on the aromatic ring of curcumin is
electron donating, it destabilizes the anion formed (from loss of
H from the phenolic -OH). As such, the -OH on the
aromatic ring of curcumin would be expected to be less acidic
than that in phenol. This serves to reinforce the concept that
both resonance and inductive effects contribute to the stability of
anions and determine the relative acidities of functional groups.
Spray with the extract from turmeric onto one of the filter paper
on which an invisible message was written with Na2CO3.
Students are usually pleased to see that a message is displayed
in red while the rest of the paper is dyed yellow. Explain that the
observations are consistent with the reported pKa value of 8.70
for curcumin corresponding to loss of its most acidic hydrogen
atom (14). Ask students what would happen to the message if it is
sprayed with HCl. Invariably, they are usually able to predict that
the message will disappear. Spray the HCl onto the paper; the
message does disappear.
Provide students with the structure of cyanidin diglucoside
and ask whether a message written with Na2CO3 will be
displayed if sprayed with a solution of the compound extracted
from roses. Many students are likely to indicate that it would.
Have them identify the most acidic proton and write resonance
structures for the anion formed. This requires them to consider
which hydrogen atom would be lost most easily and the relative
contributions of the various resonance structures. The exercise
also provides students with practice drawing more challenging
resonance structures. Spray the rose extract onto the paper; a
message is displayed in blue. The results are consistent for the
reported pKa value of 5.4 for cyanidin corresponding to loss of its
most acidic hydrogen atom (12). In conclusion, hold the filter
paper up high and spray with HCl. The message disappears.
Instructors so inclined may take a bow.
Conclusions
This demonstration can be quickly, easily, and safely prepared. We have found the demonstration to be effective in
attracting students' attention and in engaging them in lively
discussions of resonance stabilization, keto-enol tautomerization, and relative acidities. The demonstration had some unintended positive consequences. Students were curious as to why
compounds have different colors and as to the applications
of pH-dependent reversible structural changes of molecules.
These opportunities were used to reinforce additional chemical
concepts and make students aware of some of the more fascinat-

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ing current and potential applications of organic chemistry in


their lives.
Acknowledgment
Gratitude is expressed to students who took Organic Chemistry I with me and participated in the demonstration. In addition,
gratitude is expressed to Ed Vitz and the checker for their helpful
suggestions, many of which were incorporated into the demonstration. Special thanks to Kirran Tiwari, who took the colorful
pictures of the messages developed by the dye solutions.
Literature Cited
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(Vitis vivifera L.) on the Concentration and Composition of
Phenolic Compounds in Grapes and Wine. Ph.D. Dissertation
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library.oregonstate.edu/dspace/bitstream/1957/3142/1/JCthesisfinal3%5B1%5D.pdf.
13. McMurry, J. Organic Chemistry, 7th Ed.; Thompson, Brooks/
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Science, 8th Ed.; Prentice Hall: Upper Saddle River, NJ, 2000; p 620.

Supporting Information Available


pH-dependent structural changes of curcumin and vitamin A; color
pictures of messages displayed by extracts from henna, turmeric, and roses;
table of pKa values of compounds used and cited in demonstration. This
material is available via the Internet at http://pubs.acs.org.

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