Synthesis and characterization of novel azo-morphine derivatives for possible use in abused drugs analysis

Article abstract of DOI:10.1016/j.ejmech.2007.03.034

A new simple and fast spectroscopic method was presented as a new marker for heroin use. Novel azo-morphine derivatives with spectroscopic absorption peaks ranging from 330-470 nm, were synthesized by the coupling of morphine (M) and 6-acetyl morphine (6-

Full text of DOI:10.1016/j.ejmech.2007.03.034

Available online at www.sciencedirect.com
European Journal of Medicinal Chemistry 43 (2008) 357e363
http://www.elsevier.com/locate/ejmech
Original article
Synthesis and characterization of novel azo-morphine derivatives
for possible use in abused drugs analysis
b
Ahmed O. Alnajjar a, , Mohamed E. El-Zaria
*
a Department of Chemistry, College of Science, King Faisal University, P.O. Box 380, 31982 Hofuf, Saudi Arabia
b Department of Chemistry, Faculty of Science, University of Tanta, 31527 Tanta, Egypt
Received 5 February 2007; received in revised form 26 March 2007; accepted 29 March 2007
Available online 14 April 2007
Abstract
A new simple and fast spectroscopic method was presented as a new marker for heroin use. Novel azo-morphine derivatives with spectro-
scopic absorption peaks ranging from 330e470 nm, were synthesized by the coupling of morphine (M) and 6-acetyl morphine (6-AM) with
freshly prepared diazonium salt of aniline hydrochloride at 0 ꢀC. However, no reaction was observed with codeine under the same reaction con-
ditions. Separation of azo dyes was performed by TLC using tetrahydrofuran and dichloromethane in the ratio 1:1. The chemical structure of the
products was established by their microanalysis, NMR, IR, UVevis, and mass spectroscopies. Electronic absorption and excitation spectra of the
dyes were measured in solvents of different polarities. The dyes exhibited positive solvatochromism, i.e., a bathochromic band shift as the sol-
vent polarity is increased. Also, the fluorescence quantum yield was sensitive to the polarity and the pH of the medium. The UVevis spectros-
copy of spiked compounds in human urine samples was also reported. The drugs (M, 6-AM and mixture of both) were coupled with freshly
prepared diazonium salt even at very low concentration of the drugs 10ꢁ9 M.
Ó 2007 Elsevier Masson SAS. All rights reserved.
Keywords: Heroin; Morphine; Abused drugs; Dyes; Azo coupling; Fluorescence; Determination
1. Introduction
Among the most common fluorescent derivatization reac-
tions are those which target reactive species such as acids, alco-
hols, and primary and secondary amines. Unfortunately,
many abused drugs (i.e., heroin, 6-AM and morphine) are ter-
tiary amines (Fig. 1) and are not compatible with the most com-
monly utilized amine reactive fluorescent dyes. These dyes
include compounds such as fluorescein isothiocyanate isomer
I (FITC) [5], 4-(4,6-dichloro-s-triazin-2-ylamino) fluorescein
(DTAF) [6], 4-fluoro-7-nitrobenzofurazan (NBD-F) [7] and 3-
(4-carboxybenzoyl)-2-quinolinecarboxaldehyde (CBQCA) [8].
Other fluorogenic reagents specifically made for derivatization
of the tertiary amine group such as the malonic acid/acetic
anhydride system [9] and the aconitic acid method [10] result
in a deteriorating effect on the fluorescence of the reaction prod-
uct. In addition the products of these reactions are unstable, light
sensitive and give many components that seem to be associated
with the reagent blank.
Heroin, which is obtained synthetically from the acetyla-
tion of morphine, has an analgesic potency two to three times
that of the parent drug and, due to the presence of two acetyl
groups, it has better penetration across the bloodebrain barrier
[1]. Heroin itself is rarely present in detectable quantities in
body fluids. The drug hydrolyses rapidly to 6-acetyl morphine
(6-AM), which in turn hydrolyses to morphine (M). Therefore,
heroin consumption can be confirmed by identifying its two
primary metabolites [2,3]. In addition, heroin is different
from most other opioids in that it has little or no affinity for
opioid receptors in the brain. The analgesic effect of the
drug is attributed to the combined effect of 6-AM and M [4].
* Tel.: þ966553922298; fax: þ96635886437.
E-mail address: anajjar@kfu.edu.sa (A.O. Alnajjar).
0223-5234/$ - see front matter Ó 2007 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2007.03.034

358
A.O. Alnajjar, M.E. El-Zaria / European Journal of Medicinal Chemistry 43 (2008) 357e363
CH3
CH3
N
NaNO2
HCl
+
N
N Cl-
NH2
NH3+Cl-
0oC
N
diazonium salt
CH3
N
H3C
N
H3CCOO
HO
OCOCH3
OCOCH3
O
O
+
N
N Cl-
+
-O
X-O
Heroin
6-AM
O
O
OR
OR
1 (E/Z) R = H, X =H
2 (E/Z) R = H, X = Na
CH3
N
CH3
N
3 (E/Z) R = COCH , X = H
3
4 (E/Z) R = COCH , X = Na
3
Scheme 1. Coupling of M and 6-AM to their azo-derivatives.
CH3O
HO
OH
O
OH
O
eluent. The constitution and purity of each of these compounds
were established by elemental analysis, NMR, UVevis, IR,
and mass spectroscopies (see Section 4 for details). However,
no reaction was found to occur with codeine under the same
reaction conditions.
Codeine
M
Fig. 1. Structures of some drugs of abuse.
Aryl and heteroaryl dyes have been attracting much attention
over the past decade because of their rapidly increasing role in
the design of advanced materials and devices [11e19]. The
use of dyes in chemistry, biology, and medicine is growing
continuously, with many new applications in the diagnosis
and treatment of disease [20e23]. The E/Z-isomerizable
N]N double bond within a conducting chain can work as
a molecular switcher, making the aryl or heteroaryl systems
promising candidates for molecular devices[16].
The NMR spectrum of 1 showed clearly that the compound
was found in two forms Z, 30% and E, 70%, which could not
be separated. On the other hand, the two forms (3Z, Rf ¼ 0.72,
35% and 3E, Rf ¼ 0.85, 42%) of compound 3 could be sepa-
rated on TLC, however, both compounds 2 and 4 were found
only as sodium salt with Rf ¼ 0.52 and 0.55, respectively.
NMR spectrum of 1 showed the resonances of the OH protons
at d ¼ 5.45 ppm which was not observed in the spectrum of
1
compound 2. Also, H and 13C NMR spectra indicated the
The present work describes the main features of the synthe-
sis of fluorescent azo-morphine derivatives, peculiarities of
their structure, and some fundamental properties. The devel-
oped method was used for determination of M and 6-AM in
spiked human urine sample through simple and fast spectro-
scopic method.
2. Results and discussion
2.1. Chemistry
The most important problem for the development of new
morphine marker compound is tedious and time-consuming
reaction steps. To meet the challenge and to introduce such
a promising family of dyes to chemistry, material science, fo-
rensic science and medicine, we have explored simple ap-
proaches to the synthesis of azo-morphine derivatives dyes.
Azo compounds were synthesized by first reacting aniline
hydrochloride with nitrous acid (HNO2) to produce an aryldia-
zonium ion. This ion can couple with a nucleophilic M or
6-AM in basic solution to produce the azo-M (phenyl-2-azo-
morphine 1, 76% and phenyl-2-azo-morphine sodium salt 2,
24%) and azo-6-AM (phenyl-2-azo-6-acetyl morphine 3,
77% and phenyl-2-azo-6-acetyl morphine sodium salt 4,
23%) in reasonable yields (Scheme 1). All compounds were
purified by thin layer chromatography (TLC) using tetrahydro-
furan (THF) and dichloromethane (CH2Cl2) in the ratio 1:1 as
Fig. 2. (a) 1H NMR spectrum of compound 2 in CDCl3 at 20 ꢀC; (b) ESI mass
spectra of compound 2 in MeOH.

A.O. Alnajjar, M.E. El-Zaria / European Journal of Medicinal Chemistry 43 (2008) 357e363
359
Table 1
Yields of azo-M and azo-6-AM dyes (1e4) and their UVevis spectral charac-
teristics in ethanol
appearance of new signals corresponding to the organic moi-
ety of each azo compound (Fig. 2). Mass spectra of all the
compounds showed molecular ion peaks corresponding to
their expected pattern of abundance ranging from 45e98%
(Fig. 2).
Compound Color (yield, %) lmax (nm) lemission (nm) 3 (l/mol cm)
1
Yellow (76)
Yellow (24)
Orange (35)
Orange (42)
Brown (23)
335
350
445
456
470
405
410
515
527
536
24 180
35 720
1707
2
The vibrational frequency of unreactive OH band n(OeH)
of M or the COCH3 band n(C]O) was not found to be
sensitive to the connection of the azo moiety with M or
6-AM. For compound 1 n(OeH) lies in the range of
3374e3376 cmꢁ1, and for compounds 3 and 4 n(C]O)
from 1710 to 1715 cmꢁ1, comparable to the frequencies
of M n(OeH) ¼ 3375e3377 cmꢁ1, and 6-AM n(C]O) ¼
1711e1715 cmꢁ1, indicating that the intra bonding of the
hetero moiety was not perturbed by substitution on the other
phenyl ring. The IR spectra of compounds 1, 3Z, and 3E
showed absorption bands within the 3510e3515 cmꢁ1 region
characteristic of the reactive OH group of the M or 6-AM
moieties. However, the IR spectra of compounds 2 and 4
clearly indicated the lack of OH group absorptions in the
wavenumber range 3500e3515 cmꢁ1 which was also con-
firmed by the disappearance of (CeO) stretching band.
3Z
3E
4
1603
1868
band observed in UV region, in the wavelength range 265e
280 nm was attributed to pep* transition within the furan
heterocyclic moiety of the compounds. The third band
observed in the UV region at 285e295 nm was assigned to
nep* electronic transition of OH groups. This assignment
was supported by the disappearance of this band in acid
medium when excitation of the n-electrons was expected to
be hindered by protonation in acetate buffer. The compounds
displayed two main broad visible bands in ethanol within
the range 330e350 nm (compounds 1 and 2) and 440e
470 nm (compounds 3 and 4). These bands were capable of
being assigned to pep* transition involving the whole elec-
tronic system of the compounds with a considerable charge-
transfer (CT) character. Such a CT originated mainly from
the phenyl moiety (phenyl azo) to the hetero moiety, i.e.,
this band was due to intramolecular CT transition. Further-
more, the CT nature of this band was also substantiated by
the spectral behavior of these compounds in buffer solutions
of varying pHs as given hereafter. The effect of solvent on
the azo-morphine dyes produced a bathochromic shift with
increasing solvent polarity.
2.2. Optical studies
The electronic absorption spectra of the reaction mixture
showed that the reaction was time dependent. In this case,
the absorption intensity of the azo band increased slowly
with time till precipitation after 15 min (Fig. 3). The electronic
absorption spectral characteristics (lmax, 3 values) of the inves-
tigated azo compounds 1e4, in ethanolic solutions are cited in
Table 1. The high 3 values pointed to the extended conjugation
between the organic moieties through azo group. The com-
pounds comprised two to three bands in the UV region and
one band in the visible region (Fig. 4). Band of the shortest
wavelength appearing in the range 205, 240, 250 nm was
best ascribed to pep* transition of the benzenoid system of
the compounds (this band is of higher intensity). The second
The picks of the fluorescence spectra of the compounds 1,
2, 3Z, 3E, and 4 in ethanol appeared at lemission values of
405 nm, 410 nm, 515 nm, 527 nm, and 536 nm, respectively,
(Fig. 5, Table 1). The emission and excitation spectra of the
dyes were measured at a concentration of c ¼ 1 ꢂ l0ꢁ9 mol/dm3
in solvents of different polarities (Fig. 6).
Fig. 3. Visible spectra of the reaction mixture of azo 6-AM in DI water at dif-
ferent reaction times.
Fig. 4. Electronic absorption spectra of 2.5 ꢂ 10ꢁ5 M of azo-morphine deriv-
atives (1e4) in ethanol.

360
A.O. Alnajjar, M.E. El-Zaria / European Journal of Medicinal Chemistry 43 (2008) 357e363
Fig. 5. Fluorescence spectra of 1 ꢂ 10ꢁ9 M of azo compounds 1e4 in ethanol.
Fig. 7. Electronic absorption spectra of 5.2 ꢂ 10ꢁ5 M of 2 in phosphate buffer
containing 4% (v/v) ethanol.
The new fluorescent dyes were also studied in buffer solu-
tions of different pHs (1e12) containing 4% (v/v) ethanol to
affect the solubility of the dyes. The bands due to localized
pep* transitions displayed slight changes, while the azo
band (CT band) showed interesting changes with pH (for ex-
ample see Fig. 7). The spectra of dyes exhibited changes in
the extinction of the azo band only within the whole range
of pH 2e12. For compound 2, the absorbance of the azo
band decreased with rise in pH while a new band was devel-
oped at longer wavelength. Clear isosbestic points were ob-
served denoting the existence of an equilibrium of the acid
base type (Fig. 7). The pH absorbance curves displayed two
clear steps indicating the establishment of two acid base equi-
libria over the pH range 4e9. Change of the pH values had
a remarkable effect on the fluorescence spectra of the dyes.
The intensity of the fluorescence decreased with decreasing
pH value of the solution due to the protonation of the dyes.
The visible band of these azo compounds acquired a blue
shift at very low pH (<4) and a red shift at high pH (>8)
values relative to the types of media. This could be explained
based on the observation, that in highly acidic media, the pro-
tonation occurring on the furan heterocyclic moiety of the
compounds enhanced its electron withdrawing ability and
hence facilitated the CT transition, i.e., low energy was re-
quired for such a transition. On the other hand, the ionization
of the OH group which took place at high pH values (>8)
increased the electron density on the phenyl moiety of M or
6-AM and consequently increased its electron releasing charac-
ter which resulted in an easier CT. This behavior confirmed the
assignment of the visible band of these compounds to an
electronic CT (tautomeric) which took place from the electron
donor phenyl coupled moiety, to the electron acceptor hetero-
aromatic moiety of M or 6-AM in acid media and vice versa.
2.3. Analytical parameters
Under the experimental conditions described (Section 4.2),
standard calibration curves of M and 6-AM were constructed
by plotting absorbance versus concentration. The method
was found to be linear in the range of 1 ꢂ 10ꢁ3e1 ꢂ 10ꢁ5 M
with a weighed regression described in Eqs. (1) and (2) for
M and 6-AM, respectively.
A ¼ 5081:2 C þ 0:0022; r ¼ 0:9998
A ¼ 5267:2 C ꢁ 0:0005; r ¼ 0:9996
ð1Þ
ð2Þ
where C is the concentration in M and r is the correlation co-
efficient. This indicates wide dynamic range within good lin-
earity. The precision and intermediate precision (within
5 days) of the proposed method were examined by running
five experiments. The relative standard deviation (RSD) was
found to be <1.12%.
2.4. Biology
The determination of M in biological samples has become
almost a routine assay in many toxicology laboratories owing
Fig. 6. Absorption spectra of 2.5 ꢂ 10ꢁ5 M of 3Z in different organic solvents.

A.O. Alnajjar, M.E. El-Zaria / European Journal of Medicinal Chemistry 43 (2008) 357e363
361
to the spread of the abuse of heroine, which is mainly biotrans-
formed into M. In this experiment, the coupling reaction was
carried out with drug-free urine sample, and with urine sample
spiked with M and 6-AM. No remarkable change was ob-
served for the drug-free urine sample as indicated by UVe
vis spectroscopy. For urine sample spiked with the M and
6-AM, a brick red color appeared at once which was measured
by UVevis giving two peaks of azo compounds, at lmax
395 nm and 580 nm corresponding to phenyl-azo-M and phe-
nyl-azo-6-AM, respectively, (Fig. 8). The extraction recoveries
were found to be >98.8% and RSD values of the recovery did
not exceed 2.5% indicating good repeatability of the adopted
method.
3. Conclusions
Fig. 8. Absorption spectra of azo reaction mixture of M and 6-AM in human
urine sample.
A novel and rapid spectrophotometric method for the deter-
mination of M and 6-AM is proposed in this paper. A general
efficient approach to the synthesis of novel promising reactive
azo-morphine derivatives has been developed. The approach is
based on the low temperature azo coupling of M or 6-AM with
diazonium salt of aniline hydrochloride in a basic medium. It
was possible to obtain suitable colored M or 6-AM compounds
with maximum absorption peaks ranging from 330 to 470 nm
by choosing the appropriate azo chromophore. All azo com-
pounds under investigation displayed two or three bands in
the UV region in different organic solvents. The first and sec-
ond bands attributed to pep* transition in benzenoid system
and furan heterocyclic moiety, respectively, whereas the third
band in UV region was assigned to nep* electronic transition.
In the visible region, all the compounds displayed two main
broad visible bands, which were attributed to intramolecular
CT transition. The resulting azo compounds were highly fluo-
rescent in most organic solvents and water. Thus, our results
show that this reaction can be considered as a marker of heroin
use.
used for pH adjustments was Thermo Electron Cooperation,
Orion 420Aþ. The UVevis data were measured on Shimadzu
1601 PC instrument. The fluorescence (excitation and emis-
sion) spectra were determined with Shimadzu RF-5301 PC
spectrophotometer: excitation slit width ¼ 10 nm, emission
slit width ¼ 10 nm. Melting points were recorded on Gallen-
kamp apparatus.
4.2. General procedure for the synthesis of
compounds 1e4
A 0 ꢀC solution of aniline hydrochloride (0.5 mmol) and
1 N HCl (1 mL) in deionized (DI) water (5 mL) was treated
with a 0 ꢀC solution of NaNO2 (0.105 g, 1.5 mmol) in DI wa-
ter (5 mL), and the mixture was stirred at 0 ꢀC for 5 min. The
resulting diazonium salt solution was poured into a 0 ꢀC solu-
tion of M or its derivative (0.5 mmol) in NaOH (0.05 g,
1.25 mmol). The mixture was stirred at 0 ꢀC for 15 min. The
precipitate was filtered off, washed with NaCl, DI water and
dried in vacuo. The products were purified by TLC using
THF and CH2Cl2 in ratio 1:1 as eluent.
4. Experimental
4.1. General
Upon storage of the azo coupling products 1e4 at ambient
temperature for several months neither change in their UVe
vis spectra nor appearance of foreign signals in their 1H
NMR spectra were observed, which provides evidence of their
stability.
All chemicals and solvents used in this study were of ana-
lytical grade. M, 6-AM$HCl and codeine were obtained from
Lipomed Inc. (One Broaway, Cambridge, MA, USA). Aniline
hydrochloride (BDH), sodium nitrite and sodium hydroxide
were purchased from chemical sources. Plate chromatography
was conducted on Sigma-Aldrich TLC plates silica gel matrix,
H ꢂ W 20 cm ꢂ 20 cm. Elemental analyses were performed
by a Perkin-Elmer 2400 automatic elemental analyzer. All
compounds gave elemental analysis within ꢃ0.4%. The mea-
surements of NMR (1H and 13C) were carried out on a Bruker
DPX 200 spectrometer. The chemical shifts d are given in ppm
relative to X ¼ 200 MHz for d (1H) (nominally SiMe4), and
X ¼ 50 MHz for d (13C) (nominally SiMe4). IR (cmꢁ1) spectra
were determined as KBr disc on a Shimadzu FTIR 8400
spectrometer. Electron spray ionization (ESI) mass spectra
were recorded on a Bruker Esquire in CH3OH. The pH-meter
4.2.1. Phenyl-2-azo-morphine (1)
Yield (0.19 g, 76%) as a yellow solid; m.p. 73e75 ꢀC;
Rf ¼ 0.75 (THF/CH2Cl2 1:1); IR nmax(KBr disc, cmꢁ1
)
3500s, 3375s (OH), 2503s (NtH), 1635s (C]C, alkene),
1616, 1510s (C]C, aromatic), 1315e1245 (phenolic, CO),
1
1282, 1091s (CeOeC); H NMR (200 MHz; CDCl3; Me4Si)
d (ppm) Z: 8.2, 7.85, 7.51, 7.44, 7.37, (5H, d, aromatic H,
J ¼ 8.9), 7.27, 7.19, 7.06 (3H, t, aromatic H, J ¼ 7.87), 5.6,
(1H, s, OH); E: 8.16, 7.57, 7.49, 7.43, 7.37, (5H, d, aromatic
H, J ¼ 9.3); 13C NMR (50 MHz; CDCl3; Me4Si) d (ppm) Z:
156.36, 155.83, 152.25, 148.75, 147.19, 135.89, 130.69,

362
A.O. Alnajjar, M.E. El-Zaria / European Journal of Medicinal Chemistry 43 (2008) 357e363
127.31, 122.43, 121.14, 117.76, 111.89; E: 154.14, 153.94,
152.12, 148.81, 147.12, 135.68, 130.57, 127.12, 122.16,
121.03, 117.69, 111.45 (aromatic C, CH); (ESI): m/z(%) 389
(90) [Mþ]; anal. calcd. for C23H23N3O3: C, 70.95; H, 7.95;
N, 10.79; found: C, 70.89; H, 7.92; N, 10.68.
(3H, t, aromatic H, J ¼ 9.72); 13C NMR (50 MHz; CDCl3;
Me4Si) d (ppm) 170.34 (C, COCH3), 179.56, 156.17,
153.08, 147.87, 147.41, 133.98, 129.12, 127.04, 121.86,
121.15, 116.42, 111.32 (aromatic C, CH); (ESI): m/z(%) 453
(67) [Mþ]; anal. calcd. for C25H24N3NaO4: C, 66.22; H,
5.29; N, 9.27; found: C, 66.11; H, 5.08; N, 9.14.
4.2.2. Phenyl-2-azo-morphine sodium salt (2)
Yield (0.063 g, 24%) as a yellow solid; m.p. 79e81 ꢀC
Rf ¼ 0.52 (THF/CH2Cl2 1:1); IR nmax(KBr disc, cmꢁ1) 3376s
(OH), 2505s (NtH), 1639s (C]C, alkene), 1620, 1515s
(C]C, aromatic), 1286, 1092s (CeOeC); 1H NMR
(200 MHz; CDCl3; Me4Si) d (ppm) 8.21, 7.97, 7.86, 7.82,
7.78 (5H, d, aromatic H, J ¼ 8.9), 7.72, 7.27, 7.15 (3H, t, aro-
4.3. Biological studies
A 500 mg Bond Elut SPE column was used for the extrac-
tion. The SPE columns were conditioned by the sequential
passage of 2 ꢂ 3 mL of methanol, 2 ꢂ 3 mL of water and
2 ꢂ 5 mL of water adjusted to pH 9.5 with NH4OH. 10 mL
of human urine sample adjusted to pH 9.5 with NH4OH was
vortex, centrifuged and applied to the SPE columns at a rate
of 1.0 mL/min. The columns were washed with 2 ꢂ 5 mL of
distilled water and left to dry for 10 min. The drugs were
eluted with a solution consisting of a single phase mixture
of dichloromethane/acetone (50:50) and collected in glass
tubes. The elution solvent was evaporated to dryness under
a nitrogen stream. The dried residues were then reconstituted
in slightly warm water, and derivatization was carried out
and then the samples were analyzed using UVevis
spectrophotometer.
matic H, J ¼ 7.95), 4.35 (1H, s, OH), 3.38e3.25 (1H, m); 13
C
NMR (50 MHz; CDCl3; Me4Si) d (ppm) 156.16, 154.78,
152.89, 148.62, 147.73, 134.97, 130.87, 126.65, 121.54,
121.35, 116.45, 113.08 (aromatic C, CH); (ESI): m/z(%) 411
(52) [Mþ]; anal. calcd. for C23H22N3NaO3: C, 67.15; H,
5.35; N, 10.21; found: C, 67.02; H, 4.98; N, 9.94.
4.2.3. Z-Phenyl-2-azo-6-acetyl morphine (3Z )
Yield (0.1 g, 35%) as an orange solid; m.p. 71e73 ꢀC;
Rf ¼ 0.72 (THF/CH2Cl2 1:1); IR nmax(KBr disc, cmꢁ1
)
3512s, 3377s (OH), 2495s (NtH), 1746s (C]O), 1642s
(C]C, alkene), 1619, 1510s (C]C, aromatic), 1321e1245s
(phenolic, CO), 1278e1086s (CeOeC); 1H NMR
(200 MHz; CDCl3; Me4Si) d (ppm) 7.88, 7.77, 7.49, 7.38,
7.06 (5H, d, aromatic H, J ¼ 9.0), 6.93, 6.81, 6.36, (3H, t, ar-
omatic H, J ¼ 12.07), 5.52 (H, s, OH); 13C NMR (50 MHz;
CDCl3; Me4Si) d 171.21 (C, COCH3), 157.49, 152.85,
152.24, 148.14, 147.36, 134.62, 129.77, 127.51, 122.23,
121.64, 116.25, 111.47 (aromatic C, CH); (ESI): m/z(%) 431
(10) [Mþ]; anal. calcd. for C25H25N3O4: C, 69.6; H, 5.8; N,
9.74; found: C, 69.23; H, 5.56; N, 9.37.
Acknowledgement
This work was supported by the grants from King Abdula-
ziz City for Science and Technology, Saudi Arabia, award #
MT-1-2. The author is grateful to Prof. Detlef Gabel, and
Dr. Mohamed E. El-Zaria, Department of Chemistry, Univer-
sity of Bremen, Germany, for providing facilities for NMR
and mass spectra measurements. Thanks are also due to the
Department of Chemistry, College of Science, King Faisal
University, Saudi Arabia, where part of this research was
conducted.
4.2.4. E-Phenyl-2-azo-6-acetyl morphine (3E )
Yield (0.12 g, 42%) as an orange solid; Rf ¼ 0.85; m.p. 78e
80 ꢀC; (THF/CH2Cl2 1:1); IR nmax(KBr disc, cmꢁ1) 3515s,
3375s (OH), 2501s (NtH), 1751s (C]O), 1644s (C]C, al-
kene), 1616, 1514s (C]C, aromatic), 1325e1247s (phenolic,
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1
CO), 1280e1089s (CeOeC); H NMR (200 MHz; CDCl3;
Me4Si) d (ppm) 7.85, 7.77, 7.49, 7.39, 7.22 (5H, d, aromatic
H, J ¼ 10.12), 7.18, 7.07, 7.03, (3H, t, aromatic H,
J ¼ 10.12), 5.55 (H, s, OH); 13C NMR (50 MHz; CDCl3;
Me4Si) d (ppm) 170.47 (C, COCH3), 157.89, 153.11,
151.87, 149.21, 148.12, 133.57, 130.12, 127.05, 121.94,
121.32, 116.41, 111.01 (aromatic C, CH); (ESI): m/z(%) 431
(10) [Mþ]; anal. calcd. for C25H25N3O4: C, 69.6; H, 5.8; N,
9.74; found: C, 69.49; H, 5.74; N, 9.62.
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