Vibrational Spectroscopy 98 (2018) 128–133

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Vibrational Spectroscopy
journal homepage: www.elsevier.com/locate/vibspec

Vibrational and structural properties of L-Alanyl-L-Phenylalanine dipeptide T

by Raman spectroscopy, infrared and DFT calculations
⁎
C.B. Silvaa, , J.G. da Silva Filhoa, G.S. Pinheirob, A.M.R. Teixeirac, P.T.C. Freirea
a
Departamento de Física, Universidade Federal do Ceará, C.P. 6030, Campus do Pici, 60.455-760, Fortaleza, CE, Brazil
b
Departamento de Física, Universidade Federal do Piaui, campus Ministro Petrônio Portella, 64.049-550, Teresina, PI, Brazil
c
Departamento de Física, Universidade Regional do Cariri, 63.010-970, Juazeiro do Norte, CE, Brazil

A R T I C LE I N FO A B S T R A C T

Keywords: A study of vibrational and structural properties of L-Alanyl-L-Phenylalanine hydrophobic dipeptide (molecular
Dipeptide formula C12H16N2O3) is reported. The L-alanyl-L-phenylalanine is formed by two hydrophobic amino acids, L-
Raman spectroscopy alanine and L-phenylalanine, through a peptide bond. Individually these compounds have many important
FT-IR properties, including increasing immunity and providing energy to muscle tissue, brain and central nervous
DFT
system (L-alanine). We have performed measurements of Raman and Fourier transform infrared spectroscopy,
between 3500-50 cm−1 and 4000-130 cm−1, respectively, at ambient conditions. To support the experimental
results calculations using the density functional theory (DFT) with B3LYP functional, 6–31 G ++ (d, p) basis
sets and the polarizable continuum model of solvatation in an isolated molecule in the zwitterionic form were
done. The assignment of the normal modes was described by the potential energy distribution analysis. In this
way, it was possible to analyze the vibrational and structural properties of L-alanyl-L-phenylalanine, allowing
the assignment of each vibrational normal mode of the molecular structure, associating them with those results
obtained experimentally.

1. Introduction
Short peptides have arisen the interest of the scientific community
around the world. Although the peptides have less complex structures
than the proteins, they exhibit important biological activities.
Hydrogels formed by self-assembly of peptides have been used in 3D
cell culture as constituents of compounds with regenerative properties
[1,2], which act in the regeneration of several tissues with low re-
generative capacity, such as bone tissue, dental tissue, skeletal tissue
and cartilage [3–6]. Some peptides have antioxidant properties and act
as pH regulator in muscle cells [7], avoiding the oxidation of organic
molecules and metals, helping in glycemic control [8] and the treat-
ment of type 2 diabetes [9], besides being used in the diet of slimming
[10].
The L-alanyl-L-phenylalanine (L-Ala-L-Phe) is a dipeptide formed by
two hydrophobic amino acid residues. The hydrophobic dipeptides
constitute a very diversified group of crystalline structure and have
been a source of stable microporous materials [11,12], which are ori-
ginated from molecular self-assembling dictated by the formation of
hydrogen bonds and by the aggregation of hydrophobic entities in the
side chains [13].
A series of works, using the most varied solvents, shows these di-
peptides tend to incorporate molecules of organic solvents into their
structure, which then act as acceptors for one of the three H atoms of
the N-terminal amino group NH3+ [14–18]. The change of solvent can
result in the formation of different structures, for example, nanotubes,
nanofilms, nanospheres, nanofibers, and materials with large channels
filled with solvent [19–21]. These structures have been used in different
areas, such as in the construction of biosensors [21], nanocarriers for
drug and gene delivery [22] and natural encapsulates [23,24]. In ad-
dition, the use of dipeptides has been tested in the fight against diseases
such as zyka virus [25], cancer [26], diabetic osteoporosis [27], HIV
[28], malaria [29], anti-tumor [30,31], and antibacterial and antifungal
agents [32].
In previously published works, crystalline structures with L-Ala-L-
Phe dipeptide was obtained with 2-propanol solvent [33], ammonium
[34], hydrochloride dehydrate [35], as well as structures interacting
with copper [26] and gold [36], although the crystalline structure of
pure L-Ala-L-Phe dipeptide is still unknown. Similar structures con-
taining the phenylalanine showed to be an effective hydrogelator, when
used with the specific solvent. In this manner the L-Ala-L-Phe is a good
candidate to gel-forming [37–40].

⁎
Corresponding author.
E-mail address: cristiano.balbino@fisica.ufc.br (C.B. Silva).

https://doi.org/10.1016/j.vibspec.2018.08.001
Received 26 February 2018; Received in revised form 25 July 2018; Accepted 4 August 2018
Available online 09 August 2018
0924-2031/ © 2018 Elsevier B.V. All rights reserved.
C.B. Silva et al.
Structural analysis and vibrational properties of dipeptides using the
density functional theory (DFT) [41,42] have been reported in the lit-
erature, being important to understanding the behavior of aggregation
of amino acid in complex structures as peptides and proteins [43–49].
The present work reports a detailed study about the structural and vi-
brational properties of L-Ala-L-Phe dipeptide (molecular formula
C12H16N2O3) including data about important torsion angles of the
molecular structure and the assignment of vibrational modes.
2. Experimental
L-Alanyl-L-Phenylalanine (L-Ala-L-Phe) dipeptide, which consists of
a white polycrystalline powder, was used as provided by Sigma-Aldrich.
The Raman spectra of L-Ala-L-Phe were performed using a triple spec-
trometer (Jobin-Yvon T64000) equipped with an N2-cooled charge-
coupled device detector (CCD). The 532 nm line of a semiconductor
laser was used as the exciting source, with an output power of 70 mW.
The spectral resolution was ∼ 2 cm−1. An Olympus microscope lens
with focal distance 20.5 mm (N.A. = 0.35) was used to focus the laser
beam on the sample at room temperature. The spectral region analyzed
covered the region from 3500 to 40 cm−1.
The Fourier Transform infrared (FT-IR) measurement was obtained
through a Bruker Vertex 70v spectrometer. The infrared spectrum was
recorded from 4000 to 130 cm−1, using the Attenuated Total
Reflectance (ATR) technique with vacuum by absorbance of radiation
to eliminate CO2 residues and humidity with a spectral resolution of
2 cm−1 and accumulating 128 scans per spectrum.
3. Calculations
Calculation and normal mode assignment of IR and Raman spectra
is widely utilized to understand organic materials like amino acids,
peptides and proteins [50–59]. The vibrational modes of L-Ala-L-Phe
dipeptide were calculated using density functional theory (DFT). For
this purpose, we used the Gaussian 09 package [60], with the B3LYP
functional [61] and 6-31++G(d,p) basis set. Solvatation was also in-
corporated for assignment by using the polarizable continuum model
(PCM). Our calculations were established for an isolated molecule in
the zwitterionic form. In general, zwitterionic states are predominantly
observed in the crystalline medium for a variety of amino acids and
dipeptides [62].
The initial molecular structure of L-Ala-L-Phe dipeptide used was
that reported by Görbitz [33], and then optimized for a minimum of
energy. The optimized molecular structure was subjected to investigate
the conformational structure by calculation of energy surface potential
(scan) of torsion dipeptide angle and, finally, to calculate the frequency
of vibrational modes for this molecule (3N-6).
Using VEDA program [63] for molecular visualization and potential
energy distribution (PED) calculations, the vibrational assignments of
the normal modes were obtained. Here we consider only values with
contributions of at least 10%. For a better correspondence between the
theoretical and experimental spectra, we made use of the scale factors
suggested by the reference [64] of 0.977 for vibrations modes under
1800 cm−1, and 0.955 for vibrational modes above 1800 cm−1.
The calculations were made in the conformer of the lowest energy.
Although the calculations had been performed in a molecule, the vi-
brations of both single molecule and molecules in the crystal are si-
milar, exceptions occur with vibrations related to part of the molecule
involved in hydrogen bond, for example, and vibrations related to the
lattice modes that obviously do not appear in the calculation of the
molecule. It is important mentioning that the possible coincidence of
the molecular conformation in the solid state with the molecular con-
formation in the gas phase or in solution has supporters and opponents;
this question remains unresolved. As can be seen in Table 2, there is a
good agreement between the calculated wavenumbers and the experi-
mental wavenumbers of the Raman and infrared bands to the internal
129
Vibrational Spectroscopy 98 (2018) 128–133
Fig. 1. Representation of molecular structure of L-Ala-L-Phe in zwitterionic
form.
modes of the Ala-Phe molecule.
4. Results and discussions
The molecular structure of L-Ala-L-Phe (illustrated in Fig. 1) has 33
atoms, molecular weight of 236.27 g/mol and molecular area of ap-
proximately 95.5 Å2. The data of positions of atomic coordinates and
the geometric parameters (bond lengths, bond angles and dihedral
angles) of the optimized molecular structure of L-Ala-L-Phe dipeptide
are listed in Tables S1 and S2 of Supplementary Material, respectively.
4.1. Potential energy surface (PES)
An important torsion angle in dipeptides deserves attention, due to
it considerably influences the conformation of peptides and proteins.
This torsion angle is formed by the alpha carbon (αC) and the beta
carbon (βC) of the first amino acid residue with the alpha carbon and
the beta carbon of the second amino acid, interchanged by amide plane
(OCNH), θ = βC1- αC1•••αC2- βC2, which defines the relative positions of
the two sides chain in the structure of dipeptide [19].
This dihedral angle is the simplest way to describe the conforma-
tional configuration of the molecule, characterizing the influence of the
inter- and intra-molecular interactions in the crystalline medium. In
order to investigate the energetic stability of different conformational
configurations of L-Ala-L-Phe, the potential energy surface (PES) cal-
culation was carried out, varying the dihedral angle θ from -180° to
+180° degrees in steps of 10°. The relative energy with respect to the
global minimum (ΔE) was plotted against the dihedral angle, as shown
in Fig. 2. In this figure, we can see a global minimum at -110° and local
minima around -150°, +20° and +150°. At room temperature the
thermal energy (kT) is approximately 2.45 kJ/mol, which corresponds
to the dashed red line in the plot of Fig. 2. We can observe two regions
above the thermal energy at room temperature, the first region between
-85° to -115° and the second region between -177° to -130° separated by
a barrier of 1.96 kJ/mol.
In the polypeptide chain the definition of protein folding depends on
the torsion angles prior to alpha carbon, called phi angle (ϕ), and on the
torsion angle after the alpha carbon, called the psi angle (ψ) [65]. The
analysis of rotation of these angles led to the identification of permitted
regions where there is no clash between atoms, and non-permitted re-
gions, where there is clash between the atoms. In Fig. 2 we also show
the correspondence between each value of ϕ and ψ with dihedral angle
θ. In this case, while the θ angle vary from −180° to + 180°, the ϕ
angle vary between −40° and 25° and the ψ angle vary between 80°
and 110°, respectively.
C.B. Silva et al. Vibrational Spectroscopy 98 (2018) 128–133

Table 2
Calculated and scaled wavenumbers and experimental Raman and IR wave-
numbers (in units by centimeter) and classification of the vibrational modes of
L-Ala-L-Phe dipeptide with PED.
ωcalc ωscale ωRam ωIR Assignment of the molecular vibrations with PED
[%]

3638 3474 ν(N2H)[100]

3546 3386 3381 νas(N1H3+)[100]
3479 3322 3255 3256 νs(N1H3+)[98]
3205 3061 3062 3063 νs(CH-ring)[93]
3202 3058 3058 3058 νs(N1H3+)[96]
3193 3049 νas(CH-ring)[93]
3186 3043 3047 νas(CH-ring)[99]
3175 3032 3035 νas(CH-ring)[90]
3170 3027 3026 3025 νas(CH-ring)[95]
3151 3009 νs(C2H3)[99]
3145 3003 3003 3006 νas(C2H3)[96]
3133 2992 2981 νas(C5H2)[98]
3109 2969 2974 2971 ν(C1H)[95]
Fig. 2. Variation of energy and dihedral angle (ψ, ϕ) with the change in di-
3084 2945 2946 2955 νs(C5H2)[95]
hedral angle θ of L-Ala-L-Phe dipeptide. 3068 2929 2933 2829 ν(C4H)[94]
3066 2928 2920 2922 νs(C2H3)[100]
1709 1670 1673 1678 ν(C = O)[63] + ν(N2C3)[19]
Depending on the molecular interactions in the crystal structure, we
1670 1632 1631 δ(N1H3+)[85]
can observe different conformations at ambient temperature. This wide 1647 1609 1603 ν(C10=C11-ring)[30]
angular range can leave the side chains practically on opposite sides. 1637 1599 δ(N1H3+)[91]
This allows the formation of crystalline microporous structures in hy- 1625 1588 ν(C = C-ring)[41]
drophobic dipeptides, with hydrophilic internal superficies and hy- 1624 1587 1583 1565 νas(CO2−)[78]
1582 1546 1555 δ(C3N1H)[55]+ ν(N2C3)[21]
drophobic external superficies [13,66]. The majority of the known di- 1528 1493 1497 1497 δ(CCH-ring)[67]
peptides has the torsion dipeptide angle θ (in absolute values) higher 1505 1470 δ(C2H3)[89]
than 135° and other considerable portion between 90° and 135°, as 1492 1458 1457 1453 δ(C2H3)[77]
shown by Gorbitz [19]. The two ranges showed in PES of torsion di- 1490 1456 sc(C5H2)[53]
1481 1447 1444 1443 sc(C5H2)[27] + δ(R)(CCH-ring)[18]
peptide angle in L-Ala-L-Phe dipeptide are in accordance with the re-
1470 1436 wagg(N1H3+)[63] + wagg(C2H3)[13]
ported analysis. 1441 1409 1398 1389 wagg(C2H3)[81]
Another important effect on dipeptide structures is the solvent used 1398 1366 1369 γ(HC1C3N2)[62] + δ(C1C2H)[14]
in the synthesis process or the crystal growth. Many of the structures 1388 1356 1357 1356 δ (N2C4H)[24] + γ(HC4C12O3)[23] +
are established by a particular solvent; sometimes the change of it leads νs(CO2−)[11]
1364 1333 1349 νs(CO2−)[51]
obtaining different forms of the same dipeptide: nanotubes, hydrogels, 1359 1328 δ(CCH-ring)[37] + δ(C5N2H)[10]
nanolayers, vesicles, thin films, among others. For this reason, we 1353 1322 ν(CC-ring)[50] + δ(CCH-ring)[20]
submitted the molecular structure of L-Ala-L-Phe to optimize changing 1346 1315 1317 1318 δ(C2C1H)[35]
the solvent in solvatation. We chose some solvents available in Gaussian 1332 1301 1290 1289 wagg(C5H2)[33]
1297 1267 1250 1264 ν(N2C3)[26] + δ(C3N2H)[20] + δ(C1C2H)[18]
code that generally are used in the growth of some crystal structure of
1261 1232 1248 1231 γ(HN2C1C2)[45] + δ(HC4N2)[26]
dipeptides: 1-propanol, 2-propanol, methanol, ethanol, 2,2,2-tri- 1223 1195 1202 ν(C5C6)[33] + δ(HC7N8)[10]
fluoroethanol and water. 1199 1171 1182 1181 δ(HCC-ring)[39] + τ(C2H2)[12]
Table 1 shows the dielectric constant of each solvent and the cor- 1196 1168 1169 1167 δ(HCC-ring)[24] + τ(C2H2)[10]
respondent torsion dipeptide angle of the most stable conformation. 1176 1149 1155 δ(HCC-ring)[78]
1145 1119 1120 1124 r(C2H3)[17] + γ(HN1C1C2)[10] +
The two conformers of lowest energy were obtained using the methanol
γout(C2N1C3C1)[11]
and water as solvents. We can still see that torsion dipeptide angle 1105 1080 1081 1088 δ(HCC-ring)[32] + ν(CC)[25]
presents a small variation with respect to the change of dielectric 1087 1062 1047 1050 ν(C4C5)[22] + ν(N2C4)[20]
constant of solvent, increasing slightly as shown in Fig. 3. 1048 1024 1029 1028 ν(C8C9)[47] + δ(C9C10C11)[13] + δ(HC7C8)
[11]
1022 998 1012 r(C2H2)[39]
1013 990 r(N1H3+)[21] + γ(HC1C2N1)[18] + r(C2H2)[15]
4.2. Vibrational analysis
1012 989 989 δ(CCC)[62] + ν[C9C10)[12]
1004 981 γout(HCCC-ring)[67] + γout (C8C9C10C11)[16]
For the study of the vibrational properties (IR and Raman) the op- 1001 978 980 978 ν(C1C2)[23] + r(N1H3+)[17] + r(C2H2)[12]
timized structure was submitted to the frequency calculation, and the 987 964 γout(HCCC-ring)[78]
theoretical Raman spectrum was converted from Raman activity to 959 937 948 942 ν(C12C4)[19]+ γout(C12C5N2C4)[13]
932 911 918 917 γout(HCCC-ring)[65]
907 886 880 876 ν(N1C1)[20] + ν(C1C3)[16]
Table 1 868 848 854 853 ν(N2C4)[17] + ν(C4C5)[10]
Solvent effect in dihedral angle θ of L-Ala-L-Phe peptide. 859 839 844 844 γout(HCCC-ring)[96]
854 834 827 827 ν(N1C1)[26] + ν(C1C2)[15] + δ(O1C16N2)[13]
Solvent Dielectric constant (ε) Dihedral angle ΔE (kj/mol)
825 806 819 811 δ(C8C9C10)[24] + ν(C5C6)[16] + δ(CO2−)[13]
C2-C1—C4-C5 (°)
775 757 761 761 δ(CO2−)[23] + γout(O1C1N2C3)[15] +
γout(O2C4O3C12)[10]
2-propanol 19.264 −133.594 6.934
760 743 743 742 γout(HCCC-ring)[48] + γout(C6C7C8C9)[10]
1-propanol 20.524 −137.392 9.998
746 729 721 718 γout(O1C1N2C3)[30]+ δ(CO2−)[12]
2,2,2-trifluoroethanol 26.726 −138.191 10.859
+γout(O2C4O3C12)[11]
Ethanol 24.852 −138.199 7.701
710 694 698 696 γout(CCCC)[53] + γout(HCCC-ring)[41]
Methanol 32.613 −138.365 0
651 636 622 621 δ(CO2−)[14] + γout(O2C4O3C12)[14]
Acetonitrile 35.688 −138.389 4.365
632 617 δ(C9C10C11-ring)[75]
Water 78.355 −147.456 0.287
(continued on next page)

130
C.B. Silva et al. Vibrational Spectroscopy 98 (2018) 128–133

Table 2 (continued)

ωcalc ωscale ωRam ωIR Assignment of the molecular vibrations with PED
[%]

615 601 ν(C1C3)[20] + δ(O1C3N2)[12] + γout(O1C1N2C3)

[12]
581 568 566 564 δ(C6C7C8)[15] + δ(C4C12N2)[15] + δ(O3C4C12)
[11]
552 539 547 548 δ(O3C4C12)[27] + δ(C1C3N2)[10]
539 527 γ(HN2C3C1)[78]
489 478 493 471 γout(C2N1C3C1)[24] + γ(C7C8C9C10)[15]
444 434 δ(C1C2N1)[26] + δ(N2C4C5)[15] +
γout(N1C1C2C3)[12]
436 426 422 δ(N2C4C5)[28] + δ(O3C12C4)[18]
416 406 407 γout(CCCC-ring)[53]
387 378 398 395 γ(N1H3+)[37] + δ(O1C3N2)[17]
367 359 353 350 δ(C1C2N1)[11] + δ(N1C1C3)[10] +
γ(N1H3+)[10]
348 340 340 γ(CO2−)[13] + δ(C5C6C7)[11]
250 244 241 270 γ(C2H3)[33]
237 232 229 228 δ(C4C5C6)[14] + γ(C10C9C8C7)[12]
225 220 210 γ(C2H3)[18] + γout(C2N1C3C1)[16] +
γout(C12C9C8C7)[11]
202 197 γ(N1H3+)[66]
170 166 168 δ(C12C4N2)[20] + δ(C1C3N2)[14] + δ(N2C4C5)
[11]
138 135 133 δ(C3N2C4)[44] + δ(C1C3N2)[10]
102 100 γ(C3N2C4C5)[56]
83 81 85 δ(C4C5C6)[34] + γout(C5C7C11C6)[33]
66 64 γ(N1C1C3N2)[47]
61 60 γ(C1C3N2C4)[42] + γ(N1C1C3N2)[18] +
γ(N2C4C5C6)[11]
34 33 γ(C4C5C6C7)[54] + γ(C1C3N2C5)[16] +
γ(O3C12C4C5)[11]
33 32 γ(C4C5C6C7)[46] + γ(C1C3N2C5)[19]
22 21 γ(N2C4C5C6)[65] + γ(C1C3N2C5)[10]

Nomenclature: γ=torsion, γout = torsion out of plane, wagg = wagging,

δ=deformation, ν=stretching, νs=symmetric stretching, νas=anti-symmetric
stretching, r = rocking, τ=twisting.

Fig. 4. Experimental and calculated (scaled) infrared spectra above and ex-
perimental and calculated (scaled) Raman spectra below of L-Ala-L-Phe di-
peptide.

where calculated bands are not present in the experimental spectrum.

This is due the small difference in the frequency of the modes between
the calculated and the experimental values; however, these few cases do
not detract the quality of the assignment as a whole.
Table 2 presents a detailed description of the assignments of all
vibrational modes of the molecule of L-Ala-L-Phe dipeptide. In addition,
the table shows the calculated (ωcalc) and scaled (ωscale) wavenumbers,
as well as the IR and Raman experimental bands associated with the
relative assignment. Using the scaled wavenumber as a reference, we
can assign molecular vibrations for each normal modes observed in the
experimental measurements of Raman and IR spectroscopy. To facil-
itate the discussion, we separated the spectral ranges in four subsec-
tions.

Fig. 3. Evolution of torsion dipeptide angle of L-Ala-L-Phe with different sol-

vents.
Raman intensity using the procedure adopted in a previous work [67].
The Fig. 4 illustrates the comparison between theoretical and experi-
mental Raman and infrared spectra. The Raman effect is due the in-
elastic scattering of light, a process that involves the molecular polar-
izability, while infrared spectroscopy is based on effects of absorption
of radiation through the electric dipole. Eventually, because the phy-
sical effects are distinct, some modes in the Raman spectrum are not
present in the IR spectrum, and some modes in the IR spectrum are not
observed in the Raman spectra. Additionally, when comparing the
theoretical and experimental spectra one notes there are some few cases
4.2.1. Spectral region from 3500 to 2800 cm−1
The vibrations in this region are characteristic of stretching vibra-
tions of the functional groups NH, NH3+, CH, CH2 and CH3. Potential
energy distribution presented a high contribution of these portions of
the molecule in the normal modes (between 90–100%). A band ob-
served in the IR spectrum at 3386 cm−1 is attributed to an anti-sym-
metric stretching of NH3+ group. Symmetric stretching of NH3+ was
situated at 3322 and 3058 cm−1 and was observed in both spectra (IR
and Raman). At 3061 cm−1, we have the symmetric stretching vibra-
tion of CH belonging to the ring of the structure, while the anti-sym-
metric stretching can be observed between 3049 and 3027 cm−1. Anti-
symmetric stretching of the CH3 group are observed at 3009 and
3003 cm−1, and the symmetric stretching of the same group is assigned

131
C.B. Silva et al.
at 3009 to 2928 cm−1. Symmetric stretching of the CH2 group is at-
tributed to the band at 2992 cm−1, while the CH2 anti-symmetric
stretching corresponds to the band at 2945 cm−1. The bands at 2969
and 2929 cm-1 are due to the CH stretching vibration. Additionally, the
CH/OH stretching region in the IR experimental spectrum contains a
couple of features (in particular the one at 3200 cm−1) which can be
understood as due the presence of water.
4.2.2. Spectral region from 1800 to 1200 cm−1
In this region, the mixed modes begin to emerge due to vibrations of
different parts/groups of the molecule. At 1670 cm−1 we observed a
band attributed to C]O stretching plus N2C3 stretching, this band is
denominated amide I and appears as the most intense in the IR spec-
trum [68], where the latter corresponds to the peptide bond. Anti-
symmetric stretching of CO2- group occurs at 1587 cm−1. At 1546 cm
−1
appears the band corresponding to the amide II, being a combina-
tion of CN and NH vibrations [68].
Symmetric stretching is observed at 1357 and 1349 cm−1.
Deformation of NH3+ group is observed between 1632 and 1599 cm−1.
The C]C stretching vibrations in the ring of the structure is observed
from 1609 to 1588 cm−1, and CCH deformations, between 1493 and
1322 cm−1. The band situated at 1546 cm-1 is related to the C3N1H
deformation mixed with N2C3 stretching; these atoms correspond to
the amide plane, being observed a very intense band in the IR spectrum,
whereas in the Raman spectrum it appears with low intensity.
Deformations of CH3 group occur between 1470 and 1447 cm−1. The
CH3 wagging vibrations are observed from 1436 to 1409 cm−1, where
we can still observe an NH3+ wagging vibration. Wagging vibration of
CH2 occurs at 1301 cm−1 and the scissoring vibrational modes of the
CH2 group are observed at 1456 and 1447 cm−1. In-plane torsions of
HCCN and HCCO and in-plane deformations of CCH and CNH are
commons in this spectral region. We also observed NC and CC
stretching vibrations between 1267 and 1195 cm−1.
4.2.3. Spectral region from 1200 to 600 cm−1
In this spectral region predominates deformation and torsional vi-
brations, and NC and CC stretchings are also observed. Deformations of
HCC belonging to the ring occur from 1171 to 1080 cm−1. The CH3
rocking vibration gives rise to the band at 1119 cm−1, while the CH2
rocking is observed at 998 and 990 cm−1, where we also observed the
NH3+ rocking vibration. Out-of-plane torsions of HCCC belonging to
the ring are observed from 981 to 694 cm−1, where also occurs out-of-
plane torsion of CCCC, CCNC, OCNC, OCOC groups from the skeleton of
the molecular structure. Deformation of CO2- group contributes to the
bands between 806 and 622 cm−1. Out-of-plane deformation of HCCC
of the ring presents contributions in the bands from 981 to 698 cm−1.
Twisting vibration modes of the CH2 group contributes in the bands at
1171 and 1168 cm−1. Rocking vibration modes (CH2, CH3 and NH3+)
are present in the bands between 1119 and 990 cm−1. The CC
stretching has contribution in bands of the 1062 - 601 cm−1 region, and
the NC stretching presents contributions in bands located at 1047, 886,
848 and 827 cm−1.
4.2.4. Spectral region from 600 to 150 cm−1
In this region predominates deformation and torsional vibrations of
the molecular skeletal of L-Ala-L-Phe dipeptide. The NH3+ torsional
vibration is observed between 378 and 359 cm−1, the CO2- torsion
occurs at 340 cm−1 and the CH3 torsion at 220 cm−1. The low wave-
number region (lower than 150 cm−1) is associated with lattice modes,
so the calculation in a single molecule is not precise for this region. In
Table 2 we furnish a tentative assignment of the modes with wave-
number lower than 150 cm−1 as obtained from the DFT calculation for
one molecule. This indicates that although most of the bands in this
region are associated with lattice modes, possibly they are coupled to
some internal modes. However, the most important point is the possi-
bility to use the bands appearing in this region to define or not the
132
Vibrational Spectroscopy 98 (2018) 128–133
occurrence of phase transitions either under pressure [53] or under
temperature [54] variations as previously reported. Admittedly, the
understanding of this spectral region helps us to realize the stability of
the structure under diverse thermodynamics parameters.
Finally, between 3200 and 1800 cm−1, the Raman and FT-IR
spectra showed the absence of bands. This is characteristic of the ma-
jority of amino acids since the units that form the molecular structure
do not have vibrations with energy in this region. So, it is expected that
similar behavior occurs in dipeptides. This absence was reproduced by
DFT calculations, showing a good agreement with experimental results.
Furthermore, assignments for other dipeptides discussed in previous
works indicate a good agreement with the present work [43,44,69,70].
As a conclusion of this section, we have furnished a complete descrip-
tion of vibrational modes of the L-Ala-L-Phe dipeptide, giving in-
formation about the normal modes of the material and linking theore-
tical calculations with experimental spectra.
5. Conclusions
In this work, the structural and vibrational properties of the mole-
cular structure of L-Ala-L-Phe dipeptide in the zwitterionic form were
investigated using the density functional theory and vibrational spec-
troscopy. Analysis of important torsion angle in the molecular structure
of dipeptides (θ=βC1αC1•••αC2- βC2) showed two regions susceptive to
low energy conformation of L-Ala-L-Phe at room temperature. The first
region, found between 85° < |θ| < 115° and the second region, found
between 177 < |θ| < 130°, are separated by a barrier of 1.96 kJ/mol.
In these angular ranges, the side chains can carry practically on oppo-
site sides, which may facilitate the formation of structures with hy-
drophilic and hydrophobic properties as occurs with some dipeptides.
Conformers of lower energy were obtained for different solvents using
the CPMC method. The calculations showed that there is a small var-
iation of energy and the torsion dipeptide angle increases subtly with
the dielectric constant of the solvent. For L-Ala-L-Phe, conformations
with lower energy were obtained using methanol and water as solvent.
Regarding the vibrational properties of the material, the calculated
wavenumbers reproduced the experimental results with good agree-
ment, allowing us to assign all normal modes of vibrations of L-Ala-L-
Phe with the respective potential energy distribution.
Acknowledgments
The authors would like to acknowledge the financial support from
the Brazilian agencies CAPES and CNPq. We also thank the CENAPAD-
SP for the use of the GAUSSIAN 09 software package and the compu-
tational facilities through the project of reference proj373. PTCF ac-
knowledges support from FUNCAP-CNPq PRONEX PR2-011-00006.01-
00/15. J.G.S.F acknowledges the Brazilian agency CAPES for the
postdoctoral program fellowship (process 1746352).
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.vibspec.2018.08.001.
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