Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range
Using micro-Raman spectroscopy, a detailed study of vibrational spectra of L-Asparagine and L-Glutamine amino acids adsorbed on aluminum foils was carried out within the frequency range 80…3500 cm⁻¹ under different excitation wavelengths. Based on a detailed analysis of Raman spectra of the above-me...
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Інститут фізики напівпровідників імені В.Є. Лашкарьова НАН України
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| Zitieren: | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range / B.O. Golichenko, V.M. Naseka, V.V. Strelchuk, O.F. Kolomys // Semiconductor Physics Quantum Electronics & Optoelectronics. — 2017. — Т. 20, № 3. — С. 297-304. — Бібліогр.: 20 назв. — англ. |
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| citation_txt | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range / B.O. Golichenko, V.M. Naseka, V.V. Strelchuk, O.F. Kolomys // Semiconductor Physics Quantum Electronics & Optoelectronics. — 2017. — Т. 20, № 3. — С. 297-304. — Бібліогр.: 20 назв. — англ. |
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| description | Using micro-Raman spectroscopy, a detailed study of vibrational spectra of L-Asparagine and L-Glutamine amino acids adsorbed on aluminum foils was carried out within the frequency range 80…3500 cm⁻¹ under different excitation wavelengths. Based on a detailed analysis of Raman spectra of the above-mentioned amino acids and data of DFT calculations of normal modes and isotopic substitution for these analytes available in the literature, the interpretation of amino acids' vibrational bands was performed. The polarized Raman spectra of the studied amino acids indicate a different ordering of polycrystalline structure in distinct spots on the sample. The most significant variations of ratios between polarized bands are principally observed for deformation vibrations of NH₂, COO⁻ , and CH₂ groups within the entire “fingerprint” range and valence vibrations of CC and CN bonds within the range 1000…1100 cm⁻¹.
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Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
297
PACS 33.20.-t, 78.30.-j
Raman study of L-Asparagine and L-Glutamine molecules
adsorbed on aluminum films in a wide frequency range
B.O. Golichenko1, V.M. Naseka2, V.V. Strelchuk2, O.F. Kolomys2
1Taras Shevchenko National University of Kyiv,
64/13, Volodymyrska str., 01601 Kyiv, Ukraine
Corresponding author: B.O. Golichenko, e-mail: desilor227@gmail.com
2V. Lashkaryov Institute of Semiconductor Physics,
National Academy of Sciences of Ukraine,
41, prospect Nauky, 03680 Kyiv, Ukraine
Abstract. Using micro-Raman spectroscopy, a detailed study of vibrational spectra of
L-Asparagine and L-Glutamine amino acids adsorbed on aluminum foils were carried out
within the frequency range 80…3500 cm–1 under different excitation wavelengths. On
the basis of detailed analysis of Raman spectra of the mentioned above amino acids and
data of DFT-calculations of normal modes and isotopic substitution for these analytes
available in literature, interpretation of amino acids vibrational bands were performed.
The polarized Raman spectra of studied amino acids indicate different ordering of
polycrystalline structure in distinct spots on the sample. The most significant variations
of ratios between polarized bands are principally observed for deformation vibrations of
NH2, COO– and CH2 groups within entire “fingerprint” range and valence vibrations of
CC and CN bonds within the range 1000…1100 cm–1.
Keywords: amino acid, L-Asparagine, L-Glutamine, vibrational spectroscopy.
Manuscript received 27.06.17; revised version received 15.08.17; accepted for
publication 06.09.17; published online 09.10.17.
1. Introduction
In the recent two decades, a quantity of investigations of
basic organics compounds forming proteins, components
of cell cytoplasm and organelles became one of the most
numerical studies in cell biology. It is particularly due to
needs of developing new medical methods (targeted
drug delivery, nanoparticles treatment, installation of
functional implants, etc.). Optical methods for direct
biological molecule detection have been explored, and
vibrational spectroscopies show great promise among
them. The vibrational spectroscopic methods of Raman
and Fourier-transform infrared (FTIR) spectroscopy
applied to study both structural and conformational
information of biological systems, including amino
acids, proteins and lipids. Detailed understanding the
vibrational structure of amino acids is thus helpful in
studying intra- and intermolecular interactions.
However, implementation of these spectroscopic
methods is not an easy task. Recently, Zhu G. et al. [1]
give an overview of measured Raman spectra of solid
amino acids and their aqueous solutions, but today, a
database, which would include the vibrational spectra of
all amino acids, measured under the same, well-defined
and reproducible experimental conditions is not exist.
In the solid state and polar media, both the amino
acids L-Asparagine (L-Asn, 2-amino-4-amidosuccinic
acid (NH3
+–CH(CO2
–)–CH2–CONH2) and L-Glutamine
(L-Gln, 2-Amino-4-carbamoylbutanoic acid (NH3
+–
CH(CO2
–)–(CH2)2–CONH2) form a zwitterionic
structure. These are amino acids with polar neutral
charged groups and are the single ones that contain
carboxamide functional group, which plays a significant
role in formation of Van der Waals interactions and
hydrogen bonds. Asparagine is necessary for functioning
of the brain and plays an important role in formation and
functioning of proteins [2]. Glutamine, unlike to
Asparagine, is conditionally essential amino acid and
plays a more significant role in synthesis of lipids,
regulation of kidneys functionality, purines synthesis
and safe circulation of ammonia in a human circulatory
system [3]. L-Asn and L-Gln side chains can form
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
298
hydrogen bonds with water molecules, peptide
backbones or other functional groups of biological
molecules that plays an important role in formation of
the secondary structure in proteins and conformational
transitions. Spectroscopic study of these amino acids is
an important way to obtain information about molecular
conformations and the nature of hydrogen bonding in
biological substances.
The only few published infrared and Raman studies
of amino acids can be found. In the experiment
performed by M. Wolpert et al. [4], infrared vibrational
spectra and band assignments for all typical amino acids
in aqueous solutions within 500…1800 cm–1 spectral
range were carried out. Implementation of Raman
spectroscopy for the solid Glutamine analysis was
investigated only in few previous studies [3, 5, 6]. А
detailed Raman study of Asparagine monohydrate
crystal was carried out by Moreno et al. in [7, 8]. By use
of IR, Raman, inelastic neutron scattering measurements
and DFT calculation, the normal modes of vibrational
spectra of pure L-glutamine in the solid state were
determined and described [9]. The polarized Raman
spectra from the microcrystalline powder of L-glutamine
were used to general assignment of the vibrational
fundamentals on the basis of the intensity changes
observed in some vibrational bands [3]. There are few
works published in the journals concerning investigation
of fundamental vibrations and molecular structure
properties of Asparagine in the solid state [2, 5, 9-12]. It
should also be noted that the vibrational spectra of
amino acids obtained by different research groups,
perhaps measured under slightly different conditions,
might be quite dissimilar.
In the literature, only the most significant and cha-
racteristic Raman bands are often discussed for each
amino acid. Also, low-frequency vibrations assignments
performed by different authors are distinguishing by
their origin – some of the bands below 200 cm–1 are de-
scribed as lattice vibrations, while the others are attri-
buted to H-bonds formed between the analyte molecules.
Thus, the lack of information and the confusing
aspects of the existing published works indicate that a
comprehensive study of the vibrational spectra of these
molecules as well as their polarized Raman spectra and
low-frequency vibrations is necessary.
Here, we report the polarized Raman spectra of L-
Asparagine and L-Glutamine amino acids in the solid
state within the spectral range 80…3500 cm–1. The
special interest of this research was to perform a
vibrational study of L-Asn and L-Gln within the
frequency range below 400 cm–1. Using the obtained
experimental data, we carried out assignments of the
observed frequencies of Asparagine and Glutamine in
the solid state, which may be used in subsequent works
of the spectroscopic analysis of interactions between
these amino acids and other molecules of biological
importance.
2. Samples and experimental technique
Our samples were prepared by deposition of drops of
analytes diluted in distilled water with the concentration
10–3 Mol/l on thin films of industrial aluminum foil.
Optical images of surface of the films are presented in
Fig. 2. Due to hydrophobness of the surface, drops with
the volume 8 to 10 µl maintained round shape while
evaporating. After it, they formed a concentric spots
with a noticeable rim. After next 3 times of deposition,
solution was spreading faster and might flow out over
the previous rim.
The Raman spectra of amino acids were measured
using Jobin Yvon T64000 spectrometer equipped with a
thermoelectrically-cooled device CCD detector with a
spectral resolution of 0.2 cm–1. 488 and 514.5 nm laser
beams (with the power ~1 mW) were focused down to a
micrometer sized spot on the sample through the
confocal Raman microscope (Olympus BX41 with a 50×
objective) equipped with a piezo-scanner. Polarized
Raman spectra were obtained using 785.0 nm radiation
from a diode-pumped 785 nm near-infrared (NIR) laser
excitation operating at ~15 mW (experimental setup
briefly described in [13]). The acquisition time of
Raman spectra was amounted about 5–10 min for each.
All Raman spectra were subjected to processing
techniques using a commercial baseline correction
process for extracting the data.
a b
Fig. 1. Schemes of molecular structure of L-Asparagine (a) and L-Glutamine (b).
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
299
Fig. 2. Optical image of the Glutamine (top) and Asparagine
(bottom) aggregates on aluminum foil.
3. Results and discussion
The vibrational spectrum of microcrystalline acids
exhibits two distinct ranges: the low frequency range
associated to lattice vibrations; the medium and high
frequency range associated with molecular vibrations.
Despite the similar structure of L-Asparagine and
L-Glutamine molecules, the number of normal modes in
their spectra considerably varies as the additional –CH2–
section of Glutamine provides opportunities for different
skeletal and complex vibrations of the molecule.
Raman spectra of studied amino acids obtained
with VIS and NIR excitation are shown in Figs. 3 to 5.
The analysis of their origin can be performed within the
low-wavenumber range (below 400 cm–1), medium-
wavenumber range (400…1800 cm–1) and high-
wavenumber range (over 2700 cm–1). The polarized
Raman spectra in Fig. 5 were obtained using the
Wollastone polarizing prism. Marking z(x, x)ž(||,||)
denotes that both polarizer and analyzer have the same
direction, whereas z(y, x)ž(⊥,||) indicates that the
analyzer has perpendicular orientation.
Low-wavenumber range 100…400 cm–1
Spectroscopic studies of Raman scattering in the
aforementioned amino acids showed the impact of
rotational spectra of Oxygen and Nitrogen molecules to
the final ones. Thus, comparing the spectra of amino
acids and clear aluminum substrate, we excluded them
as well as edge-filter artifacts.
Raman peaks that appeared within the low-fre-
quency range of amino acids shown in Fig. 3 and enlis-
ted in Table 1 include the bands attributed to lattice vi-
brations and molecular vibrations involving the COO–
torsion, NH3
torsion and CCαN deformation ones [6-
9, 14-16]. They are specific for each type of molecules.
The Raman bands of amino acids within the lattice vi-
brational range below 150 cm–1 arise as a result of the
rotational and translational vibrations of the molecules
or their parts in crystal, however Pawlukojc et al. [9] as-
signed the L-Glutamine bands detected within this range
as torsion vibrations of CO2
– unit and molecule back-
bone. Different types of intermolecular interactions
caused by the forces between hydrocarbon molecules are
generally weak, and therefore do not contribute signi-
ficantly to the spectrum.
100 150 200 250 300 350 400
1
2
L-Asparagine
τ(
N
H
3+ )
δ(
C
O
N
H
2)
δ
(C
C
C
)
δ(
C
C
α
N
)
τ
(C
C
)
τ(
C
O
O
- )
δ(
C
C
)+
τ(
C
C
)
In
te
ns
ity
, a
rb
.u
n.
Raman shift, cm-1
×0.1
3
100 150 200 250 300 350 400
1
2
3
δ
(C
C
C
)
δ(
C
C
α
N
)
τ
(C
C
)
τ(
C
O
O
- )
δ(
C
C
)+
τ(
C
C
)
In
te
ns
ity
, a
rb
.u
n.
Raman shift (cm-1)
L-Glutamine
4
a b
Fig. 3. Low frequency Raman spectra of L-Asparagine (a) and L-Glutamine (b) amino acids adsorbed on aluminum films at T =
293 K and excitation wavelengths: 785 nm (⊥,||) (1) and (||,||) (2), 488 nm (3) and 514 nm (4).
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
300
Table 1. Vibrational frequencies and band assignment for the low-frequency Raman spectra of the L-Asparagine and
L-Glutamine amino acids recorded at different excitation wavelengths and polarization geometry (here and after, we
use the following abbreviations: s = strong, m = medium, w = weak, v = very, sh = shoulder, sc = scissoring, r =
rocking, wag = wagging, tw = twisting, ν = stretching, δ = bending, γ = out-of-plane bending, τ = torsion, s =
symmetric, a = antisymmetric).
L-Asparagine L-Glutamine
488 nm 785 nm (||,||) 785 nm (⊥,||) 488 nm 514 nm 785 nm (||,||) 785 nm (⊥,||)
Assignment
124 m 124 m 126 w 126 w
131 m 131 m 131 m 133 m 135 m 136 w 136 m
Lattice vibrations
147 m 147 m 147 m
162 m 162 m 162 m
δ(CC) + τ(CC)
187 m 189 m 187 m 184 m 185 m 185 m 186 m τ(COO–)
211 m 211 m 213 m 213 m
237 w 237 w 235 w 238 m 239 w 240 m 242 w
292 w 293 w 293 w 291 m 291 w 294 m 294 m
338 m 337 m 337 w
350 m 351 w 350 vw 343 m 343 w 343 m 343 m
δ(CCC);
δ(CCαN);
τ(CC)
392 m 392 w 392 w δ(CONH2)
402 m 403 w 404 w τ(NH3
+)
The bands at 147 and 162 cm–1 of L-Asn were
attributed to bending vibrations of skeletal structure, a
band at 188 cm–1 originates from COO– torsion
vibrations. The mode at 131 cm–1 in Asparagine as well
as modes at 125 and 135 cm–1 in Glutamine can be
assigned to molecular associated vibrations in amino
acid in the solid state [3]. These bands belong to B-
symmetry modes and should be both Raman and IR
active [7]. The medium intensity Raman band of L-Gln
located at about 185 cm–1 was attributed to a COO–
torsion vibration; the medium bands at 212, 240, 293
and 343 cm–1 were attributed to skeletal vibrations of
torsion and bending nature. Similar features are
represented by L-Asn at 236, 293, 337 and 350 cm–1.
Also, bending and torsion vibrations of Asparagine
functional groups were identified at 382 and 403 cm–1.
Medium-wavenumber range 400 – 1700 cm–1
From Figs. 4 and 5 one can see that Raman spectra of L-
Asn and L-Gln within this range have a large number of
bands corresponding to bending vibrations of the NH3
+
and stretching vibrations of the CO2 and C=O groups,
symmetric and antisymmetric vibrations of molecular
backbone and side-groups. Vibrational frequencies
obtained from the measured micro-Raman spectra and
proposed assignments for the observed bands are
summarized in Table 2. Most assignments are based on
the works [3, 7], while other sources were usually
indicated in text.
The spectra of studied amino acids may be conditi-
onally separated into several ranges, where the signi-
ficant types of vibrations are observed. Within the range
450…700 cm–1, one can find the complex vibrations
involving massive parts of the molecule. It should be
noted that symmetrical stretching vibrations of CO2
–
group in both amino acids have a similar value in Raman
spectra (566 cm–1 for Asparagine and 563 cm–1 for
Glutamine). Meanwhile, the NH3
+ torsion mode appears
in Glutamine at 478 cm–1 and at 496 cm–1 in Asparagine.
This assignment was made using the previous reports for
Alanine (NH3
+ torsion mode found at 477 cm–1 in
[17, 18]) and cysteine (at 498 cm–1 in [19]). The NH2
torsion vibration gives rise to the band at 537 cm–1 [11],
as compared to the bands at ∼520 cm–1 (Asparagine) and
∼544 cm–1 (Glutamine) observed in our work (Fig. 4).
The modes that include complex full-molecule
bending were situated between 600 to 670 cm–1 for both
– Asparagine and Glutamine [3, 18]. Further, we see the
100 cm–1 gap in Raman spectra of both analytes, typical
for many organic substances.
From 700 to 1400 cm–1, there is a range of
stretching and bending vibrations of C–C bonds as well
as out-of plane deformation ones related to NH2, CO2
–
and CH2 groups. In between 775…800 cm–1 range, we
observe rocking and twisting vibrations of side parts;
different rocking vibrations of CH2 group were identified
at 886 and 896 cm–1 for Asparagine and Glutamine,
respectively. The remaining asymmetrical bending
vibrations of CH2, NH2, and NH3
+ units are mainly
observed around 1200 cm–1.
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
301
450 600 750 900 1050 1200 1350 1500 1650
L-Asparagine
L-Glutamine
785 nm
448 nm
448 nm
785 nm
In
te
ns
ity
, a
rb
.u
n.
Raman shift, cm-1
2900 3000 3100 3200 3300 3400
νs(NH2)
νa(NH3
+)
νa(NH2)
ν(
O
H
)
ν a(N
H
2)
νa(NH3
+)
νs(CH2)
ν a(C
H
2)
ν s(C
H
)
In
te
ns
ity
, a
rb
.u
n.
Raman shift, cm-1
L-Glutamine
L-Asparagine
ν s(C
H
2)
ν a(C
H
2)
ν s(C
H
)
ν s(C
H
2)
a b
Fig. 4. Raman spectra of L-Glutamine and L-Asparagine in (a) medium-wavenumber (“fingerprint”) and (b) high-wavenumber
range (λexc = 488.0 nm). T = 293 K. Within high-wavenumber range, Raman spectra were normalized on the band at 2934 cm–1.
450 600 750 900 1050 1200 1350 1500 1650
⊥
L-Asparagine
L-Glutamine
In
te
ns
ity
, a
rb
.u
n.
Raman shift, cm-1
|| ||
||
⊥
|| ||
||
Fig. 5. Polarized Raman spectra of L-Asparagine and L-Glutamine within the wavenumber range 200…1700 cm–1 with
z(x, x)ž(||,||) and z(y, x)ž(⊥,||) scattering geometries. λexc = 785 nm. T = 293 K.
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
302
Table 2. Vibrational frequencies and bands assignment for the Raman spectra of L-Asparagine and L-Glutamine
within the medium-wavenumber spectral range.
L-Asparagine L-Glutamine
488 785 (||,||) 785 (⊥,||) 488 514 785(||,||) 785
(⊥,||)
Assignments
430 430 430 430
452 454 455 455
δ(skel)
496 494 496 477 477 480 480 τ(NH3
+)
519 520 520 541 543 545 545 τ(NH2)
567 566 566 561 561 564 564 δs(CO2
–)
603 603 602 623 623 625 625 δ(NH3
+) + δs(CONH2)
663 663 663 654 654 653 653 γ(CCN) [2] / δa(CONH2) [7] / δa(COO–) [14]
798 798 798 777 778 778 778 δr(CH2) [3, 15, 16] / δtw(NH2) [9]
824 824 826 807 807 808 γ(CO2
–)
837 837 836 γ(NH2)
848 849 850 850 ν(CC)
885 886 886 896 896 896 896 δr(CH2)
911 910 910 925 925 925 925 νs(CC)
1006 1004 1004 1001 1001 1001 1001
1052 1053 1051 1051
1086 1087 1089 1088
ν(CC) [3, 9]
1075 1074 1076 1097 1097 1098 1098 ν(CN) [3]
1101 1101 1101 1105 1105 1103 1105 δr(NH2)
1144 1143 1143 1134 1135 1133 1133 δr(NH3
+)
1151 1151 1151 1164 1167 1164 1164 δr(NH3
+) [3] / δr(NH2) [7]
1235 1235 1235 1204 1204 1205 1203 δtw(CH2) [3] / δwag(CH2) [9]
1261 1261 1261 1261 δtw(CH2) [3] / δr(CH2) [9]
1284 1286 1283 1283 δwag(CH2)
1300 1299 1299 1309 1309 1308 1310 δwag(NH2)
1331 1332 1331 1331
1360 1359 1359 1358 1359 1356 1356
δ(CH)
δs(CH)
1399 1398 1397 δs(CH2)
1404 1404 1402 1418 1418 1417 1418 δ(CN) AIII
1426 1426 1425 1428 1428 1425 1425 δs(CO2
–) [3] / δsc(CH2) [9]
1439 1437 1437 1450 1451 1449 1449 δsc(CH2)
1498 1498 1496 1497 δs(NH3) [3, 9]
1552 1552 overtone (2×776) [9]
1591 1588 1588 1588 ν(NH2) AII
1595 1594 1592 1605 1606 1605 1604 δa(NH3) [3, 9] / H2O [7]
1632 1628 1624 1624 1624 1625 νa(CO2
–) [3, 9] / δ(NH2) [7]
1642 1642 1642 1646 1648 1648 1648 ν(C=O) [7] / δa(NH3) [3, 9]
1688 1690 1689 1689 ν(C=O) AI [3, 9]
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
303
The range 1050–1100 cm–1 contains C–C stretching
modes of Glutamine backbone, which are absent in
Asparagine. The characteristic Amide III features of
Asparagine and Glutamine were identified at 1075 and
1098 cm–1, respectively. A much lower frequency
position of this band in the spectrum of Asparagine may
be related to distinctive allocation of functional groups
and, therefore, their different relative dispositions to
backbone.
Within the range after 1400 cm–1, the spectra
mainly contain the asymmetrical stretching and bending
vibrations of functional and sidechain groups. They also
contain scissoring CH2 vibrations (1426 and 1438 cm–1
for Asparagine, and 1427 and 1450 cm–1 for Glutamine).
Unlike to Glutamine, Asparagine spectrum demonstrates
no features between 1450 to 1550 cm–1.
A comparison between vertically and perpen-
dicularly polarized spectrum components that have been
recorded simultaneously was performed for Raman
spectra normalized on bands at 910 cm–1 (for
Asparagine) and 92cm–1 (for Glutamine). There is only
one band in Asparagine spectra at 1628 cm–1 that was
absent at perpendicular polarization. Many other modes
show insignificant deviation in intensity levels between
differently polarized components. The bands at 337, 520,
798, 886, 1073 and 1235 cm–1 are much larger in case of
parallel polarization, while the intensity of bands at
1001, 1143, 1359 and 1426 cm–1 in perpendicularly
polarized spectrum prevail over their equivalents with
lesser values.
For Glutamine, we carried out more detailed
polarization study. The spectra were obtained from 6
different spots and then averaged. It was found for
modes at 480, 925, 1088, 1204, 1261, 1425, 1497,
1605 and 1624 cm–1 minimal difference in their
intensities under different polarizations of incident
laser beam. The intensities of modes at 185, 213, 343,
625, 778, 1001, 1284 and 1404 cm–1 in the averaged
spectra are comparable, but may slightly deviate
within 20…50% range for each separate
measurement. Meanwhile, some bands of Glutamine
spectrum are regularly polarized. The polarization
ratio between the parallel and perpendicular
components of the mode at 545 cm–1, which
corresponds to the torsion bending of NH2 group,
typically is ½. Similar ratios were found for the bands
at 850, 1098, 1331 and 1418 cm–1 (⅓, ⅓, ¼ and ¾,
respectively). The modes at 1098 and 1418 cm–1 are
related to C–N bond stretching and bending vibrations
and, therefore, reveal distinct polarization ratio.
In the spectra of Glutamine, we also observed the
inversely polarized modes. The band at 241 cm–1 related
to complex backbone bending was usually absent in
parallel polarized spectrum. The symmetrical bending
and asymmetrical stretching feature of CO– unit located
at 564 cm–1 were regularly polarized with the ratio 4/3.
The intensity of perpendicularly polarized component of
653 cm–1 mode was very weak or up to 4 times smaller.
Similar behavior was found for the band at 1588 cm–1
that corresponds to Amide II stretching vibration. The
polarization ratio of asymmetrical bending mode at
1449 cm–1 was found to be 2/1. The band corresponding
to the Amide mode at 1689 cm–1 was weakly
distinguishable, so that its polarization ratio remains
uncertain.
The only difference between the experimental
spots, where the Raman spectra were obtained, was
aluminum film morphological variability and random
nature of amino acids recrystallization in solution.
Taking these factors into account, we can state, that the
microaggregates of Asparagine and Glutamine obtained
from water solutions reveal complex disposition on the
surface that affects their optical properties.
High-wavenumber range 2700…3500 cm–1
Within this range, the vibrations associated with C–H,
N–H and O–H stretching modes are expected to appear
(Fig. 4b). Its assignments were identified according to
the references [2, 3, 7-9, 11, 20]. The Raman spectrum of
microcrystalline L-Asp shows three bands occurring at
the highest frequencies, which are associated with the
modes involved in hydrogen bonds formation:
asymmetric amino group stretching vibrations νа(NH2)
(at 3386 cm–1), asymmetric ammonium group stretching
νa(NH3) (at about 3113 cm–1) and the weak band at
3402 cm–1, often associated with OH symmetric
stretching vibration of water νs(OH) [2, 20], however, it
may be related to some OH vibration of the coupled
oxygen and hydrogen from carboxyl and ammonium
group of the same molecule, because of a reasonably
large difference between experimental and DFT-
calculation data [2]. In the Raman spectrum of L-Gln,
we assigned the bands at 3404 and 3175 cm–1 to the
asymmetric and symmetric stretching of the amino
group [3, 9]. The band at 3212 cm–1 corresponds to the
asymmetric stretching mode of NH3
+ unit.
Within the high-wavenumber range, the spectrum
of L-Glutamine reveals more intensive bands of CH
modes that are related with the presence of one
additional –CH2– segment in this amino acid. It is
noticeable not only for the enhanced intensity of CH2
symmetric and asymmetric stretching, but also for
appearance of CH mode at 2991 cm–1. The study of
Dhanelincourt et al. [3] states that the significant
difference in intensity of polarized spectra of Glutamine
is mostly observed for the band at 2950 cm–1 (which we
found at 2952 cm–1) for its in-plane attaching to NH2
group. The significant decrease in the polarized
spectrum intensity was also found for the bands
corresponding to both symmetrical and asymmetrical
stretching modes of amino groups. In the work of
Moreno et al. [7], polarized electric field mostly affects
the intensity of bands at about 3113 and 3402 cm–1,
which is consistent with our assumption about the nature
of L-Asparagine stretching mode at 3402 cm–1.
Semiconductor Physics, Quantum Electronics & Optoelectronics, 2017. V. 20, N 3. P. 297-304.
doi: https://doi.org/10.15407/spqeo20.03.297
© 2017, V. Lashkaryov Institute of Semiconductor Physics, National Academy of Sciences of Ukraine
304
4. Conclusions
This study shows a significant difference in the Raman
spectra of similar polar uncharged amino acids L-
Asparagine and L-Glutamine, molecular structures of
which distinguish only by the backbone methyl group.
This difference influences the CH-vibrations frequency
range as well as the fingerprint range due to spatial
reorganization of functional groups. The dissimilar
crystalline structure also affects their polarization
features, which, we believe, depends on crystallization
of aggregates. It could be seen from experiments that the
intensity levels of vibrational bands, which correspond
to normal modes of functional molecular groups,
discriminate mostly.
The results presented in this report clearly show
that the technique involving a micro-Raman
spectroscopy provides good quality, reproducible
vibrational spectra of amino acids in solid state. The
spectra can be used to identify an amino acid or to
elucidate the nature of adsorption of the molecules upon
metallic surfaces.
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|
| id | nasplib_isofts_kiev_ua-123456789-214955 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1560-8034 |
| language | English |
| last_indexed | 2026-03-21T11:33:50Z |
| publishDate | 2017 |
| publisher | Інститут фізики напівпровідників імені В.Є. Лашкарьова НАН України |
| record_format | dspace |
| spelling | Golichenko, B.O. Naseka, V.M. Strelchuk, V.V. Kolomys, O.F. 2026-03-05T12:05:37Z 2017 Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range / B.O. Golichenko, V.M. Naseka, V.V. Strelchuk, O.F. Kolomys // Semiconductor Physics Quantum Electronics & Optoelectronics. — 2017. — Т. 20, № 3. — С. 297-304. — Бібліогр.: 20 назв. — англ. 1560-8034 PACS: 33.20.-t, 78.30.-j https://nasplib.isofts.kiev.ua/handle/123456789/214955 https://doi.org/10.15407/spqeo20.03.297 Using micro-Raman spectroscopy, a detailed study of vibrational spectra of L-Asparagine and L-Glutamine amino acids adsorbed on aluminum foils was carried out within the frequency range 80…3500 cm⁻¹ under different excitation wavelengths. Based on a detailed analysis of Raman spectra of the above-mentioned amino acids and data of DFT calculations of normal modes and isotopic substitution for these analytes available in the literature, the interpretation of amino acids' vibrational bands was performed. The polarized Raman spectra of the studied amino acids indicate a different ordering of polycrystalline structure in distinct spots on the sample. The most significant variations of ratios between polarized bands are principally observed for deformation vibrations of NH₂, COO⁻ , and CH₂ groups within the entire “fingerprint” range and valence vibrations of CC and CN bonds within the range 1000…1100 cm⁻¹. en Інститут фізики напівпровідників імені В.Є. Лашкарьова НАН України Semiconductor Physics Quantum Electronics & Optoelectronics Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range Article published earlier |
| spellingShingle | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range Golichenko, B.O. Naseka, V.M. Strelchuk, V.V. Kolomys, O.F. |
| title | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range |
| title_full | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range |
| title_fullStr | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range |
| title_full_unstemmed | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range |
| title_short | Raman study of L-Asparagine and L-Glutamine molecules adsorbed on aluminum films in a wide frequency range |
| title_sort | raman study of l-asparagine and l-glutamine molecules adsorbed on aluminum films in a wide frequency range |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/214955 |
| work_keys_str_mv | AT golichenkobo ramanstudyoflasparagineandlglutaminemoleculesadsorbedonaluminumfilmsinawidefrequencyrange AT nasekavm ramanstudyoflasparagineandlglutaminemoleculesadsorbedonaluminumfilmsinawidefrequencyrange AT strelchukvv ramanstudyoflasparagineandlglutaminemoleculesadsorbedonaluminumfilmsinawidefrequencyrange AT kolomysof ramanstudyoflasparagineandlglutaminemoleculesadsorbedonaluminumfilmsinawidefrequencyrange |