Kraft Reactin-Dye Synthesis
Kraft Reactin-Dye Synthesis
Kraft Reactin-Dye Synthesis
This procedure is adapted from: J.V. McCullagh, K.A. Daggett, J. Chem. Ed. 2007, 84(11),
1799.
Pre-lab Questions:
OH HO O OH
O
H2SO4 (cat)
2 + O
O
OH
Fluorescein
O
O
OH (Et)2N O N(Et)2
O
H2SO4 (cat)
2 + O
O
N(Et)2
Rhodamine B
O
O
Friedel-Crafts reactions are a class of EAS reactions that involve reaction of an aromatic
ring with an electrophile that bears an electron-deficient carbon center. There are two variations
on this reaction. One variant involves generating a carbocation from an alkyl chloride and a
Lewis acid catalyst (alkylation). The other involves an acyl halide or anhydride and a Lewis acid
catalyst to form an aromatic ketone (acylation). Protic acids can also be used to generate
carbocations. In this experiment, a Bronsted acid catalyst (sulfuric acid) will be used to generate
xanthene dyes from substituted phenols and phthalic anhydride. Two successive Friedel-Crafts
First Friedel-Crafts:
OH O O OH
H2SO4 (cat)
+ O
OH CO2H OH
O
O H O H OH
H O
O
O H O
O O O
OH
O
Acylium ion
O OH O OH
+
CO2H OH CO2H OH
The first step in the formation of fluorescein is a Friedel-Crafts acylation reaction. In this
reaction, the acyl cation is generated by protonation of the phthalic anhydride as shown above.
The acyl cation will then react with the substituted phenol (resorcinol). This produces a
substituted benzophenone.
The reaction does not stop at this point. Under the reaction conditions used, the
substituted benzophenone formed in the first reaction is protonated to form a cationic
intermediate that can participate in a reaction similar to the Friedel-Crafts alkylation reaction.
The resulting cation then undergoes alkylation to complete the assembly of the triarylmethane
carbon skeleton. Subsequent acid catalyzed esterification (lactonization) then yields flourescein
(next page).
OH
H
OH O CO2H
HO
OH HO COOH
H
HO
OH
HO HO
OH OH
HO
H
O COOH O COOH
H
Proton transfer
OH H2O OH
HO HO
OH
Intramolecular
Fischer esterification
O
O
O
HO Fluorescein
Light in the ultraviolet and visible regions can cause an excitation of electrons within a
molecule. The electrons move from bonding or nonbonding orbitals into higher energy
antibonding orbitals. For many organic compounds, these relatively high energy transitions
correspond to light in the UV region leading to uncolored compounds. In general, increasing the
amount of conjugation in a molecule will cause the energy gap between the highest occupied
bonding orbital and the lowest unoccupied * antibonding orbital to decrease. This results in a
shift in absorbance to lower energies (longer wavelength) with increasing amounts of
conjugation. If a sufficient amount of conjugation is present, absorbance of light will fall in the
visible range of the spectrum leading to colored organic compounds.
The xanthene dyes synthesized in this experiment all are sufficiently conjugated to absorb
in the visible range of the spectrum provided they are in a form where the central carbon between
all three aromatic rings is sp2 hybridized. This allows conjugation between the different aromatic
rings. We can see this if we consider the dye fluorescein. Fluorescein has been shown to have at
least 3 reasonably stable neutral tautomers, each of which have different physical characteristics.
This has been demonstrated by the fact that 3 distinct crystalline forms of fluorescein with
different colors (red, yellow and colorless) can be formed depending on how the crystals were
HO O O HO O OH HO O OH
OH O O
O O O
Fluorescence
At a 90 angle to the light source against a non-reflective surface, like a black lab bench,
the solution will appear to be green. The fluorescence properties of fluorescein can also be seen
if we illuminate a dilute solution of the compound with a black light (long wave UV lamp). If
fluorescein or rhodamine B is synthesized, their fluorescent properties will be observed as part of
the experiment.
1. Synthesis of Fluorescein:
The sand bath temperature for the first step of this reaction should be between 180 and
200C. To a large test tube (15 X 150 mm) add 153 mg of resorcinol and 100 mg of ground
powdered phthalic anhydride. To this mixture of powders add 3 drops of 4N H2SO4 (DO NOT
ADD MORE THAN 3 DROPS). Stir the mixture briefly with a spatula. Place the test tube in
To the test tube add 5 mL of acetone and 1.27 cm X 0.79 cm stir bar. Using a ring stand
and clamp, place the test tube over a magnetic stir plate and stir the solution for 5 to 10 minutes.
The solution should turn yellow as the crude fluorescein dissolves. If the entire product did not
dissolve, repeat the process with an additional 5 mL of acetone until the entire product dissolves
(do not use more than 15 mL total). Combine the acetone layers in a 50 mL round bottom and
remove the solvent on a rotary evaporator. Take this crude residue and dissolve it in 20 mL of
diethyl ether and 1 mL of water. (Note: Even though most of the dye will end up in the organic
layer it will not dissolve unless a small amount of water is present.) Place a stir bar in the
solution and put the round botttom over a magnetic stir plate for several minutes until all the
solids dissolve. Using a small separatory funnel extract the organic layer once with 10 mL
water. Following this, extract the ether layer once with 10 mL of a saturated NaCl (brine)
solution. Dry the organic layer over anhydrous sodium sulfate, filter, and remove the solvent on
a rotary evaporator to yield the product as an orange solid. Weigh the crude product before
proceeding to part 3.
2. Synthesis of Rhodamine B
The sand bath temperature for the first step of this reaction should be between 180 and
210C. To a large test tube (15 X 150 mm) add 228 mg of 3-(diethylamino)-phenol and 100 mg
of ground powdered phthalic anhydride. To this mixture add 3 drops of 4N H2SO4 (DO NOT
ADD MORE THAN 3 DROPS). Briefly stir the mixture with a spatula. Place the test tube in a
preheated sand bath deep enough so that its contents are just slightly (0.5 cm) below the surface
of the sand. The reaction should be run at a temperature between 180 and 210C. Periodically
adjust the heater setting to keep the temperature within this range. (Note: It is extremely
important to monitor the temperature and keep it within this range. Overheating will cause the
product to decompose.) Start timing the reaction once it reaches 180C. The reaction should
run for 1 hour within this temperature range. The reaction should become red-violet in color.
To the test tube add 3 mL of methylene chloride and a 1.27 cm X 0.79 cm magnetic stir
bar. Using a ring stand and clamp, place the test tube over a magnetic stir plate and stir the
solution for 5 to 10 minutes. Remove the dye solution and set it aside. Repeat this process with
additional 3 mL of methylene chloride until the entire product dissolves. This should take about
a total of 12 mL. Using a small separatory funnel, extract the organic layer once with 10 mL
5% sodium bicarbonate solution. Following this, extract the organic layer twice with 10 mL of
water (THINK: Which layer is on top?) Dry the organic layer over anhydrous sodium sulfate,
filter, and remove the solvent on a rotary evaporator to yield the product as a dark magenta
colored solid. The product may be slightly tacky at this point due to residual solvent trapped in
the product. Proceed to part 3.
3. Observation of Fluorescence
Next observe the same solution at a position 90 degrees from the light source. The color
observed from this perspective is primarily due to fluorescence.
Finally, in a darkened room shine a long wave UV lamp at the sample vial. The vials
should visibly glow from the fluoresced light. Record your observations, then proceed to step 4.
4. IR
Take an IR of a portion of your remaining crude sample. Submit the rest to your TA in a labeled
vial.
Post-lab Questions:
1. Give an explanation of why fluorescein is orange in acetone and nearly colorless
in ether.
2. Based on the functional groups observed in the IR, which form of your xanthene
dye predominates in your IR spectrum?