A rapid synthesis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid: Experimental, DFT study and DNA cleavage activity

Article abstract of DOI:10.1016/j.molstruc.2018.06.032

The newly synthesized 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid has been prepared by diazotization of anthranilic acid and coupling with 2-amino-4,6-dimethylpyrimidine. Its structure has been characterized by spectroscopic measurements (¹H NMR spectra, FT-IR spectra, mass spectra and UV–visible spectra) and thermal analysis technique. The DNA cleavage activity of compound is evaluated by agarose gel electrophoresis with a series of concentrations. Our measurements show that neither a disruptive effect created by 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid on pBR 322 DNA are observed, nor the dependence of the concentration on the activity of newly synthetized chemical on pBR 322 plasmid DNA such as cleavage or break DNA double helix structure. Moreover, computational chemistry method based on Density Functional Theory (DFT) employing B3LYP level with 6-31G(d) basis set has been used to study geometry and spectroscopic properties such as FT-IR and UV–vis spectra of the titled compound considered in this work. The computations of the chemical shifts for ¹H NMR of the title compound have been carried out via Gauge-Invariant Atomic Orbital (GIAO) method utilizing the same basis set. It is observed that DFT results are compatible with the experimental results.

Full text of DOI:10.1016/j.molstruc.2018.06.032

Accepted Manuscript
A rapid synthesis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid:
Experimental, DFT study and DNA cleavage activity
Çiğdem Karabacak Atay, Fatih Duman, Merve Gökalp, Tahir Tiliki, Sevgi Ozdemir
Kart
PII:
S0022-2860(18)30733-6
10.1016/j.molstruc.2018.06.032
MOLSTR 25319
DOI:
Reference:
To appear in: Journal of Molecular Structure
Received Date: 14 March 2018
Revised Date: 8 June 2018
Accepted Date: 8 June 2018
Please cite this article as: Çğ. Karabacak Atay, F. Duman, M. Gökalp, T. Tiliki, S. Ozdemir Kart, A rapid
synthesis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid: Experimental, DFT study and
DNA cleavage activity, Journal of Molecular Structure (2018), doi: 10.1016/j.molstruc.2018.06.032.
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ACCEPTED MANUSCRIPT
A rapid synthesis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid:
Experimental, DFT study and DNA cleavage activity
Çiğdem KARABACAK ATAY1, Fatih DUMAN2, Merve GÖKALP3, Tahir TİLKİ3*, Sevgi OZDEMIR KART4
1Mehmet Akif Ersoy University, Education Faculty, Elementary Education Department, 15030, Burdur, Turkey
2Erciyes University, Faculty of Science, Biology Department, 38039, Kayseri, Turkey
3Süleyman Demirel University, Faculty of Science & Arts, Chemistry Department, 32260, Isparta, Turkey
4Pamukkale University, Art and Science Faculty, Department of Physics, 20020, Denizli, Turkey
tahirtilki@sdu.edu.tr
Abstract
The newly synthesized 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid has
been prepared by diazotization of anthranilic acid and coupling with 2-amino-4,6-
dimethylpyrimidine. Its structure has been characterized by spectroscopic measurements (1H-
NMR spectra, FT-IR spectra, mass spectra and UV-visible spectra) and thermal analysis
technique. The DNA cleavage activity of compound is evaluated by agarose gel electrophoresis
with a series of concentrations. Our measurements show that neither a disruptive effect created
by 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid on pBR 322 DNA are
observed, nor the dependence of the concentration on the activity of newly synthetized
chemical on pBR 322 plasmid DNA such as cleavage or break DNA double helix structure.
Moreover, computational chemistry method based on Density Functional Theory (DFT)
employing B3LYP level with 6-31G(d) basis set has been used to study geometry and
spectroscopic properties such as FT-IR and UV-vis spectra of the titled compound considered
in this work. The computations of the chemical shifts for 1H-NMR of the title compound have
been carried out via Gauge-Invariant Atomic Orbital (GIAO) method utilizing the same basis
set. It is observed that DFT results are compatible with the experimental results.
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Keywords: Anthranilic acid, Heterocyclic dye, Spectroscopic property, pBR322 DNA
cleavage, Density Functional Theory.
1. Introduction
Anthranilic acid contains carboxyl (-COOH) and amino (-NH2) groups which is a precursor to
the amino acid tryptophan. Because of its medicinal and biological properties, researchers have
motivated to study anthranilic acid and its derivatives [1-4]. Moreover, biological–medical
studies of azo dyes such as anticancer [5], antitumor [6], antifungal [7], antioxidant [8] and
antibacterial [9] has been extensively studied. Therefore, the synthesis of azo dyes with
anthranilic acid takes important notice of not only scientifically but also technologically.
High yielding and clean synthesis of azo dyes make them to be significant compounds.
Although, several properties of azo dyes have been widely investigated, there is limited study
about DNA cleavage properties of azo dyes. S.M. Pradeepa et. al. [10] have synthesized Cu(II)
and Co(II) complexes of azo-containing Schiff base and investigated their DNA photo cleavage
abilities by agarose gel electrophoresis. The presence of azo and carboxylic acid group
complexes in the compound reveals the efficient DNA photo cleavage activity. Carla T. Mapp
et al. [11] have synthesized the symmetrical carbocyanine dyes and evaluated their DNA photo
cleavage activities. They have reported that the irradiation at 575, 588, 623, or 700 nm
produces good photo cleavage of plasmid DNA.
Because of the biological importance of azo dyes and anthranilic acid, we have carried out the
synthesis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid with anthranilic
acid and 2-amino-4,6-dimethylpyrimidine and evaluated their DNA cleavage activity for a
series of concentrations. The results obtained from this study may be useful for the usage of
azo dyes and further cancer studies.
The computational chemistry methods based on DFT are useful tools to determine some
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characteristic properties of the chemical and biological molecules [12-14]. Computational
chemistry methods can be used to investigate the molecular structure, thermodynamic
properties, frontier molecular orbitals, molecular electrostatic potential, non-linear optical
properties, fundamental vibrational modes and NMR spectra for small and large sized chemical
molecules [15-29]. They are used to investigate structural, vibrational and NMR properties of
the dithiophosphonates [28,29], calix[4]arenes [15,30], conduction polymer [31] and azo dyes
[32-34].
Experimental method for the synthesis and characterization of the molecule being taken into
account in this work are presented in Section 2. The computational procedure followed in this
work is given in Section 3. Both experimental and theoretical results for the structural and
vibrational properties of the compound considered in this study are given in Section 4. The
results obtained from DFT method are compared with the experimental data in the same
section. The summary and conclusion arising from this work are given in the last section.
2. Experimental
2.1. Synthesis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid
20 mmol of anthranilic acid is dissolved in hydrochloric acid:water (1:1). The solution is then
cooled to 0-5oC by stirring. While stirring, sodium nitrite (2 mmol) in water (10 ml) is
gradually added to this solution. The reaction mixture is stirred for 2h at 0-5 oC. The resulting
diazonium salt solution is then added dropwise to a cooled and stirred solution of 2-amino-4,6-
dimethylpyrimidine (20 mmol) in sodium acetate (4 g) dissolved in 20 mL ethanol:water (1:1).
Stirring is continued for 4h at 0-5 oC. The precipitated products diluted with cold water, filtered
off, washed with water several times, and dried. The obtained product is recrystalized from
DMF-H2O mixture (Orange crystal, melting point: 240 oC decomposition).
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The general route for the synthesis of 2-((2-amino-4,6-dimethylpyrimidine-
5yl)diazenyl)benzoic acid is shown in Figure 1.
2.2. DNA cleavage experiments
pBR322 plasmid DNA is purchased from commercially (Thermo Fischer Scientific, SD0041).
Cleavage effect of newly synthetized chemical 2-((2-amino-4,6-dimethylpyrimidine-
5yl)diazenyl)benzoic acid on pBR 322 plasmid DNA is monitored performing gel
electrophoresis experiment. Cleavage studies are conducted by following the procedure given
in the study of Duman et al. [35] with some modifications. Briefly, 3 ꢀL of plasmid DNA (0.5
ꢀg/mL) are incubated with 30 ꢀL aliquots of decreasing concentrations of 2-((2-amino-4,6-
dimethylpyrimidine-5yl)diazenyl)benzoic acid from 100 µM to 20 µM at 37 °C during
overnight in a buffer solution containing DMSO. After that, 10 ꢀL aliquots of chemical/DNA
mixtures are loaded onto the 1% agarose gel with loading buffer (2 µL bromophenol blue dye).
The gel is run at 80 V for 4 h in a TAE buffer (40 mM Tris acetate/1 mM EDTA, pH 8). After
electrophoresis, the gel is subsequently stained by ethidium bromide (0.5ꢀg/mL). At last, the
bands observed are visualized under UV light and photographed with a video camera.
3. Computational Method
DFT calculation utilizing B3LYP level with 6-31G(d) basis set has been carried out by using
Gaussian 09W [36] program to predict the molecular structure and vibrational frequencies of
the title compound. B3LYP level is the three-parameter hybrid method, which is the Lee–
Yang–Parr gradient-corrected correlation functional (LYP) [37] and coupled with Becke's
three-parameter gradient exchange correlation functional (B3) [38]. The three dimensional
optimized structure of the title compound obtained from GaussView program [39] is shown in
Figure 2. The equilibrium structural parameters of the title molecule are used to calculate the
vibrational frequencies, the chemical shifts and UV-vis spectrum. The vibrational
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wavenumbers of the molecule have been assigned by combining the results of the GaussView
5.0.8 program and the potential energy distribution (PED) obtained from VEDA4 program
1
[40]. Theoretical H and 13C NMR isotropic shielding calculations of title molecule are
1
13
obtained via the Gauge-Invariant Atomic Orbital (GIAO) method [41]. The H and C NMR
chemical shift calculations are investigated by considering Tetramethylsilane (TMS) as a
reference. UV-vis spectra of the title compound are also studied via Time Dependent Density
Functional Theory (TD-DFT) utilizing B3LYP level with the 6-31G(d) basis set in the different
solvents.
4. Results and Discussion
4.1. Molecular Geometry
The geometric structure of compound, synthesized in this work, predicted from DFT/B3LYP/6-
31G(d) level are presented in Figure 2 along with the atom numbering scheme. The geometry
of the compound possesses C1 point group symmetry. This compound has 33 atoms and has got
93 fundamental vibrational modes.
4.2. Structural Properties
Some important structural parameters such as bond lengths, bond angles and dihedral angles,
obtained by using the DFT/B3LYP/6-31G(d) method, are given in Table 1.The bond lengths of
O20-H33, N17-H31, O19-C18 and N8-N7 for the compound are calculated as 0.9757
1.0079 , 1.2139 and 1.2661 , respectively. The bond angles of C10-N11-C12 and N8-N7-
C4 for the title compound are found as 117.28 and 112.95 . The dihedral angles of N8-N7-C4-
C5 and N13-C14-C9-N8 are predicted as -30.04 and -180.89 . All data describing the
Å,
Å
Å
Å
º
º
º
º
compound studied in this work are given in Table S1 as a supplementary data in Appendix A.
33 bond lengths, 53 bond angles and 67 dihedral angles are necessary in order to define the
molecular structure of the compound. These bond lengths, bond angles and dihedral angles are
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given in the Table S1 as shown in Figure 2. To the best of our knowledge, there is no
experimental data on the geometric structure of the compound in the literature.
4.3. FT-IR spectra, 1H-NMR spectra, mass spectra and UV-Vis Spectra
The FT-IR spectra of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid shows
aromatic (Ar–H) band at 3200 cm−1, aliphatic (Alip–H) band at 2640 cm−1 and azo (N=N) band
at 1640 cm−1. FT-IR spectra of compound and transmittance versus wavenumber of compound
computed from DFT/B3LYP method with 6-31G(d) basis set are illustrated in Figure 3. It is
observed that the title molecule has 93 normal modes; 32, 31 and 30 modes of vibrational
modes are stretching vibrations, bending vibrations and 30 torsional vibrations, respectively.
This molecule has also 30 modes of CH vibrations. The vibrational frequencies computed from
DFT/B3LYP/6-31G(d) level are multiplied by a scale factor of 0.9613 to compare the
experimental frequencies, because the DFT method tends to overestimate the vibrational modes
[42]. Deficiencies of DFT method are due to the insufficiencies of the theoretical
approximations used in the calculations. Vibrational frequencies evaluated from DFT/B3LYP
level with 6-31G(d) basis set are given in Table 2 along with the experimental data. All of the
wavenumbers of the title compound are calculated to be positive and this case supports that
title molecule is a true minimum on the potential surface. The theoretical data are in agreement
with the experimental data except for some wavenumbers. GaussView 5.0.8 and VEDA4
programs are used to assign the vibrational normal modes of the molecule synthesized in this
work. Figure 4 shows the linear regression between theoretical and experimental data. Linear
equation of ꢀ = ꢁꢂ + ꢃ is used to perform the linear regression. Here, ꢁ and ꢃ in the equation
are fit constants. The linear regression equality of ꢀ = 0.9792ꢂ − 16.221, (ꢄ =0.988), is
obtained. It is shown that the theoretical frequencies computed from DFT/B3LYP level with 6-
31G(d) basis set are consistent with those of the experiment, since the slope of the linear
regression value (ꢄ =0.988) goes to unity as shown in Figure 4.
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1H-NMR spectra of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid shows
broad peaks at 8.01 ppm (NH2, pyrimidine) and 11.97 ppm (-COOH) of anthranilic acid. The
other δ values of 7.92–7.05 ppm (aromatic H), 2.47 ppm (CH3) of pyrimidine are recorded. 1H-
NMR spectra of compound is shown in Figure 5. 1H-NMR values calculated by
DFT/B3LYP/6-31G(d) for the compound show broad peaks at 4.25 and 4.28 ppm (NH2,
pyrimidine) and 5.54 ppm (-COOH) of anthranilic acid. The other δ values of 7.44–7.17 ppm
1
(aromatic H), 2.62-1.62 ppm (CH3) of pyrimidine are predicted. H-NMR experimental data
and theoretical values calculated from DFT/B3LYP/6-31G(d) method are given in the Table 3.
As shown in the Table 3, except for X-H, the experimental data of aromatic and aliphatic
protons are consistent with the results of the theoretical results. Different results seen in X-H
can be explained as tautomerization and resonate of X-H protons at different regions.
Theoretical 13C-NMR resonances at 141.74, 117.83, 115.93, 114.81 and 108.42 ppm are
obtained and assigned to phenyl carbons (C4, C2 and C6, C3, C1 and C5). Moreover, the
resonances of 161.49, 146.45, 146.26 and 124.78 ppm are computed and defined to pyrimidine
carbons (C14, C10, C12 and C9). 155.79 ppm is also identified to carboxylic acid carbon (C18)
and 20.42 and 15.36 ppm are dedicated to methyl carbons (C16 and C15). The 13C-NMR
values calculated from DFT/B3LYP/6-31G(d) method are given in Table 4.
Mass spectra are recorded as MS: (m/z, 100 eV): 272 [M+1]+. A mass spectrum of compound
is illustrated in Figure 6.
UV-spectra analysis of 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid have
been investigated in eight different solvents of dimethyl formamide, methanol, acetic acid,
chloroform, acetonitrile, tetrahydrofurane, dichloromethane and dimethyl sulphoxide.
Absorption spectra are determined for a serious of concentrations (10−6–10−8 M) between 300-
700 nm. A single maximum with a shoulder is observed in all solvents except for acetic acid
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and acetonitrile. The UV–vis spectra show that the bathochromic shift of λmax values is greater
for chloroform than that for the other solvents. The shoulders values predicted in all the
solvents are very similar. Experimental absorption spectra of the compound are compared with
the corresponding DFT Uv-vis results in the Figure 7. Upon examined the UV results obtained
from DFT method, λmax values of compound do not show remarkable change for all solvents
used. The solvent effects on λmax (nm) obtained by both the experimental measurement and
theoretical methods are given in Table 5. Experimental and theoretical λmax (nm) values for the
different solvents are compatible with each other, as shown in Table 5.
To analyze the thermal stability of the compound, thermogravimetric analysis (TGA) is
performed. Thermal stability of compound is illustrated in Figure 8. The absence of weight
o
loss up to 100 C indicates that the solids have humidity and water molecule. The curve of
o
TGA analysis shows approximately 24% weight loss in the temperature range of 100-316 C,
which represents the presence of adsorptive solvent molecules and the initial decomposition
temperatures (Td) is 316oC. Consequently, we can say that the compound exhibits good thermal
stabilities up to initial decomposition temperature Td of 316 oC.
4.4. Interaction with pBR322 plasmid DNA and DNA cleavage
DNA cleavage studies have been conducted extensively to examine the potential effect of
newly synthesized compound [43-45]. When the electrophoresis behavior given in Figure 9 is
examined, two different bands can be seen clearly. First band, namely supercoiled Form I, has
faster migration rate than the second band called as open circular Form II. That is, bands can be
divided by each other under electrical charge. Each column in Figure 9 refers the behavior of
electrophoresis as a function of concentration of 2-((2-amino-4,6-dimethylpyrimidine-
5yl)diazenyl)benzoic acid matter. Any change in the electrophoresis has not observed as the
concentration increases. On the other hand, it has been reported in the work conducted by Redy
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et. al. [46] that some compounds of benzocoumarin-pyrimidine hybrids show DNA cleavage
activity and they can be used as antibacterial agent. In this study, we have applied 20 µM, 40
µM, 60 µM, 80 µM and 100 µM concentrations but we do not observe any significant effect on
pBR 322 DNA even if we have applied highest concentration (100 µM). In toxicological
studies, it is not possible to observe any change without reaching a certain threshold. So, no
change was observed as we were not able to reach the threshold value for the conditions we
have set for our work.
5. Conclusion
We have synthesized 2-((2-amino-4,6-dimethylpyrimidine-5yl)diazenyl)benzoic acid in this
1
work. It is characterized by some spectroscopic studies such as H-NMR spectra, FT-IR
spectra, mass spectra, UV-visible spectra and thermal analysis technique. DNA cleavage
activity of compound is also evaluated by means of agarose gel electrophoresis technique for a
series of concentrations. Our results show that the 2-((2-amino-4,6-dimethylpyrimidine-
5yl)diazenyl)benzoic acid interaction with DNA is weakly for the concentrations applied.
When using this synthesized compound for health applications, this trait should be taken into
consideration. Additionally, DFT calculations are used to prove the molecular structure and
1
some spectroscopic properties of the title compound. The chemical shift calculations of H-
NMR and 13C-NMR spectra are performed via Gauge-Invariant Atomic Orbital (GIAO)
method. The positions of hydrogen and carbon atoms of molecule are verified by means of the
13
1H and C-NMR chemical shifts computed. UV-vis spectrum analysis of the title compound
has been investigated by both experimental technique and TD-DFT method. To our knowledge,
spectroscopic and structural investigations of the title compound are performed experimentally
as well as theoretically for the first time in this work. It is concluded that the theoretical results
are in good agreement with the experimental data when the comparison between the observed
and corresponding the calculated values. Hence, the theoretical data have approved that the
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DFT theory with B3LYP/6-31G(d) level calculations is powerful tool to investigate the
structural and vibrational properties of title compound.
Acknowledgments
The authors are grateful to SDU-BAP (Project No. 4575-YL2-16) for their financial support.
Appendix A. Supplementary material
Supplementary data associated with this article can be found, in the online version, at http://
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Figure Captions
Figure 1. General route for the synthesis of compound.
Figure 2. The structure of compound optimized via DFT/B3LYP level with the basis set of 6-
31G(d).
Figure 3. a) Measurement of FT-IR spectra of compound. b) Transmittance versus
wavenumber of compound computed from DFT/B3LYP method with 6-31G(d) basis set.
Figure 4. The linear regression between the experimental and theoretical frequencies of
compound.
Figure 5. 1H-NMR spectra of compound.
Figure 6. Mass spectra of compound (experimentally).
Figure 7. Absorption spectra of compound a) Experimental, b) Theoretical computed from TD-
DFT/B3LYP level with 6-31G(d) basis set.
Figure 8. Thermal analysis of compound (experimentally).
Figure 9. Cleavage of pBR322 DNA by different concentrations of 2-((2-amino-4,6-
dimethylpyrimidine-5yl)diazenyl)benzoic acid matter. K. Control, 1. 20 µM, 2. 40 µM, 3. 60
µM, 4. 80 µM and 5. 100 µM.
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Figure 1.
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Figure 2.
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Figure 3.
a)
b)
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Figure 4.
3500
3000
2500
2000
1500
1000
500
y = 0.9792x - 16.221
R² = 0.988
0
0
500
1000
1500
2000
2500
3000
3500
Experimental (cm-1)
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Figure 5.
8.2 8.0 7.8 7.6 7.4 7.2 7.0 6.8 6.6
f1 (ppm)
14.5
13.5
12.5
11.5
10.5
9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
f1 (ppm)
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Figure 6.
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Figure 7.
a)
0.4
0.2
0
Acetic Acid
DMF
DMSO
Kloroform
Asetonitril
THF
Dichloro
methane
300
400
500
600
700
Wavelength (nm)
b)
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Figure 8.
Temperature (oC)
24

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Figure 9.
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Table Captions
Table 1. The structural parameters computed from DFT/B3LYP/6-31G(d) level in the
optimized ground state structure of compound; Bond length (Å), bond angle (o) and dihedral
angle (o).
Table 2. FT-IR experimental data and theoretical values obtained from DFT/B3LYP/6-31G(d)
method.
1
Table 3. H-NMR experimental data and theoretical values calculated from DFT/B3LYP/6-
31G(d) method.
Table 4. Theoretical 13C-NMR values calculated from DFT/B3LYP/6-31G(d) method.
Table 5. The solvent effects on λmax (nm) obtained by both experimental measurement and
theoretical methods.
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Table 1.
Parameters via Gaussian
Bond Length (Å)
DFT/ B3LYP/6-
31G(d)
0.9757
R (20,33)
R (17,31)
R (17,32)
R (19,18)
R (8,7)
O20-H33
N17-H31
N17-H32
O19-C18
N8-N7
1.0079
1.0079
1.2139
1.2661
R (10,11)
R (13,14)
R (11,12)
R (13,12)
R (17,12)
R (8,9)
N11-C10
N13-C14
N11-C12
N13-C12
N17-C12
N8-C9
1.3334
1.3306
1.3479
1.3482
1.3560
1.3930
R (20,18)
R (7,4)
O20-C18
N7-C4
1.3572
1.4136
Bond Angles (º)
C10-N11-C12
H33-O20-C18
H31-N17-C12
H32-N17-H31
019-C18-O20
N8-N7-C4
N13-C14-C9
N11-C12-N13
C12-N13-C14
N17-C12-N11
Dihedral Angles (º)
N8-N7-C4-C5
N13-C14-C9-N8
N11-C12-N13-C14
C14-C9-N8-N7
O20-C18-C5-C4
A(10,11,12)
A(33,20,18)
A(31,17,12)
A(32,17,31)
A(19,18,20)
A(8,7,4)
A(13,14,9)
A(11,12,13)
A(12,13,14)
A(17,12,11)
117.28
105.52
118.75
120.13
122.30
112.95
121.75
127.07
116.25
116.41
D(8,7,4,5)
D(13,14,9,8)
D(11,12,13,14)
D(14,9,8,7)
-30.04
-180.89
-0.60
-192.38
-51.40
D(20,18,5,4)
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Table 2.
Vibrational frequencies (cm-1)
DFT/B3LYP*
Experimental
νAlip-H
νAr-H
3200
νN=N
νAr-H
νAlip-H
νN=N
2640
1640
3080 (C6-H24)
3069 (C1-H21)
3069 (C2-H22)
3080 (C3-H23)
2946 (C15-H25,26,27)
2949 (C16-H28,29,30)
1442
*basis set 6-31G(d), scale factor 0.9613 [42]
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Table 3.
Experimental
Alip-H
DFT/B3LYP
Aro-H
X-H
Aro-H
Alip-H
X-H
7.05-7.92 (m. 4H) 2.47 (pyrimidine,CH3) 8.01 (pyrimidine, NH2)
11.97 (-COOH)
7.17-7.44 (m. 4H)
2.62 (H27)
2.47 (H29)
2.19 (H28)
2.11 (H25)
1.79 (H30)
1.62 (H26)
4.25 (N17-H31)
4.28 ( N17-H32)
5.54 (H33)
29