Lecture 7a

Synthesis of Coumarins
R

R' O O
 Introduction I
• Coumarin (2H-chromen-2-one) was first isolated from the tonka bean
(coumarou) or sweet clover in 1820 by A. Vogel. It has a sweet odor
that is recognized as the scent of a new-mown hay and a bitter taste. O O

• Since 1882, it has been used in perfumes and as aroma enhancer in

pipe tobacco and some alcoholic drinks (i.e., mulled wines).
However, it is banned as food additive in the United States due to
the concerns of its hepatotoxicity (=chemical-driven liver damage). OH HO

• Many coumarin derivatives like dicumarol, warfarin and

acenocoumarol act as anticoagulant and vitamin K antagonist. O
O O
O

• Umbelliferone (7-hydroxyl-coumarin), found in carrots, coriander

OH
and garden angelica.
• Hymecromone (7-hydroxy-4-methylcoumarin, 4-methyl-
umbelliferone), the compound synthesized in the lab, is used in bile
O O

therapy. It is also used in the fluorometric determination of enzyme

activity. HO O O
 Introduction II
• Coumarin itself has been used as an edema modifier. Like other
benzopyrones, it is known to stimulate macrophages to degrade extracellular
albumin, allowing faster resorption of edematous fluids.
• Umbelliferone absorbs ultraviolet light strongly at several wavelengths,
which led to its use as a sunscreen and an optical brightener for textiles
as well as gain medium for dye laser. It can also be used as a fluorescence
indicator for metal ions such as copper and calcium.
• W. H. Perkin, the discoverer of the aniline dyes, first synthesized coumarin
in 1868 by the reaction of salicylic aldehyde and acetic acid anhydride in
the presence of a base.
CHO O O
NaCH3COO
+
O
OH O O
 Mechanism I
• In 1884, H. v. Pechmann developed a condensation reaction using a phenol and
a carboxylic acid or ester containing a b-carbonyl group to afford a coumarin.
• The reaction can be regarded as an esterification or transesterification followed
by the attack of the activated carbonyl group in ortho position to the oxygen atom
attached to the ring.
• The final step is a dehydration that leads to the formation of an additional double
bond in the newly formed six-membered ring.
OH O O O O O O O O
+H+ -H+
+
-EtOH
EtO R O OH OH

R' R' R' R'

transesterification R R
enolization R

+H+
R’=m-OH
electrophilic attack
dehydration
O O O OH O OH O OH
-H+ +H+ -H+
- H 2O

R' R' R' R' H

R R OH2+ R OH R OH

• Alternatively, a mechanism that starts with an attack of the ring to the keto group, which
is followed by a transesterification to form the six-membered ring has been proposed.
 Mechanism II
• The reaction is usually catalyzed by sulfuric acid, phosphoric acid,
aluminum trichloride or trifluoroacetic acid. Unfortunately, many of
these reactions require large quantities of the catalyst and elevated
temperatures.
• More recently, bismuth chloride (5 % mol, 75 oC), iron(III) chloride
in ionic liquids, montmorillonite K-10 or iron(III) fluoride under CH3

solvent-free conditions have been used successfully. In addition,

Keggin-type heteropolycompounds (i.e., H3PW12O40) have been
used under solvent-free conditions. R' N
H
O

• The reaction works better for donor-substituted benzene ring 4-methyl-2-hydroxy-quinolines

(i.e., polyhydroxy, amino, etc.). In some cases, a polycondensation

is observed particularly if an excess of ethyl acetoacetate was used. O

• If amines are reacted with ethyl acetoacetate, 4-methyl-2-hydroxyl-

quinolines are formed via acetoacetanilides (using p-TSA/MW) or R' N CH3
2-methyl-4-hydroxyquinolines via ethyl b-anilinocrotonates. H

2-methyl-4-hydroxy-quinolines
 Experimental I
• In a 50 mL Erlenmeyer flask, • Why is a small container used
resorcinol, ethyl acetoacetate and here?
Amberlyst A15 are mixed thoroughly The reaction is run in micro-scale,
which means that only little liquid
is formed upon heating
• The flask is covered with a watch
glass and is then heated in the
microwave • The reaction
Which settingshould be runhere?
is used at 30 %
power, which is equal to ~210 W
• After removing the reaction mixture,
it is allowed to cool slowly for 30
minutes • The
How canis the
flask cooling
isolated process
with paper towels
slowed
and down?
aluminum foil
• A minimal amount of hot solvent
(methanol:water=5:1)
is added The compound dissolves well in hot
• methanol but mixture
Why is the less it the cold
usedsolvent
here?
mixture
 Experimental II
• The catalyst is removed from the • How is this accomplished?
hot solution The bulk of the solution can be decanted
• Upon cooling, the product and the rest be transferred with a pipette
precipitates as a white solid
• The crystals are isolated by • What is used to rinse the crystals?
vacuum filtration in a Hirsch
First, the ice-cold solvent mixture is used
funnel and rinsed
and then some ice-cold methanol
• After sucking air through the solid
for 5-10 minutes, the compound is
dried under the heating lamp (or in
an open container until next lab
meeting).
• A sample for GC/MS analysis is • What is the proper concentration
submitted here? 2 mg/mL in ethyl acetate
 Characterization I
• Fluorescence
• Stage 1: Excitation
• A photon of energy hνEX is supplied by an external source such as an incandescent lamp
or a laser and absorbed by the fluorophore, creating
an excited electronic singlet state (S 1‘).
• Stage 2: Excited-State Lifetime
• The excited state exists for a finite time (typically 1–10 ns). During this
time, the fluorophore undergoes conformational changes and is also subject to a
multitude of possible interactions with its molecular environment.
• The energy of S1‘ is partially dissipated, yielding a relaxed singlet excited state (S 1)
from which fluorescence emission originates.
• Not all the molecules initially excited by absorption return to the ground state (S 0) by
fluorescence emission. Other processes may also depopulate S 1.
 Characterization II
• Stage 3: Fluorescence Emission
• A photon of energy hνEM is emitted, returning the fluorophore to its ground state S 0. Due to energy dissipation during
the excited-state lifetime, the energy of this photon is lower, and therefore of longer wavelength, than the excitation
photon hνEX.
• The difference in energy or wavelength represented by (hνEX – hνEM) is called the Stokes shift (dashed line: excitation,
solid line: emission). The Stokes shift is fundamental to the sensitivity of fluorescence techniques
because it allows emission photons to be detected against a low background, isolated from excitation photons.
• In contrast, absorption spectrophotometry requires measurement of transmitted light relative to high incident light
levels at the same wavelength.

• The beverage contains quinine, which absorbs at l=350 nm and emits at l=460 nm (bright blue/cyan hue)
350000 Fluorescence Spectrum of Hymechromone in MeOH

300000

250000

200000

150000

100000

50000

0
300 320 340 360 380 400 420 440 460 480 500
 Characterization III
• Fluorescence Test
• Some of the product is dissolved in 95 % ethanol
• Tube A: 1 mL of 1 M sodium hydroxide solution is added
• Tube B: 1 mL of 2 M HCl is added
• Tube C: add nothing to this test tube
• How do the solutions look like at this point?
• Place all three tubes in the UV-light (long wavelength,
365 nm). How the solutions look like now?
• Warning: Do not look straight into the UV-lamp!
 Characterization IV
• Crystal Structure (7-hydroxy-4-methylcoumarin)
• 7-hydroxy-4-methylcoumarin is almost planar but the hydroxyl
derivative displays a more than 100 oC higher melting point than
coumarin.
• The higher melting point is due to the formation of hydrogen bonds
between the phenolic hydroxyl group and the oxygen atom of the
carbonyl group of a neighboring molecule (d(O-H…O=C=189 pm),
which is similar in length compared to water.
 Characterization V
• Infrared Spectrum (7-hydroxy-4-methylcoumarin)
• n(OH)=2400-3400 cm-1
• n(C=O)=1673 cm-1
• n(C=C)=1596 cm-1
• n(CO)=1067, 1134 and 1267 cm-1
• d(oop, tri-subst.alkene)=838 cm-1

n(OH)

d(oop)

n(C=O) n(CO)
n(C=C)
 Characterization VI
• 1H-NMR Spectrum (in DMSO-d6)
• d=10.5 ppm (OH, H1)
• d=6.8 ppm (d, H2) and 7.5 ppm (d, H3) (aromatic protons)
• d=6.7 ppm (s, H6) and 6.1 ppm (s, H5) (aromatic and alkene proton)
• d=2.3 ppm (CH3, H4)

3 4
H CH3

H H
2 5

H2O in
HO O O
1
H3 H2 DMSO H4
H
6 H6 H5
H1 DMSO
 Characterization VII
• 13C{1H}-Spectrum
• Ten signals total
• Five quaternary carbons
• One carbonyl function (C9, 161.6)
• Three ipso-carbons (C1 (160.7),
C4 (112.5), C5 (154.0))
• One disubstituted alkene (C7, 155.3)
• Four methine carbons H 10 CH3

• Three CH (C2 (113.3), C3 (127.1), H 2

3

4
7

8
H

C6 (102.6)) HO 1 5 O 9 O
6
• One CH (C8, 110.7) H

• One methyl carbon (C10, 18.6)

 Characterization VIII
• Mass Spectrum
• m/z=176 ([M]+)
• m/z=148 ([M-28]+): loss of one carbon monoxide molecule
leading to a benzofuran
• m/z=147 ([M-28-1]+): loss of one carbon monoxide molecule O
HO
and a proton
• m/z=120 ([M-28-28]+): loss of two carbon monoxide molecules