Romanian Biotechnological Letters
Copyright © 2012 University of Bucharest
Vol. 17, No.5, 2012
Printed in Romania. All rights reserved
ORIGINAL PAPER
The Structures of Hydrazine Crystal and Its Antibacterial Activities
Received for publication, January 10, 2012
Accepted, June 11, 2012
FU QIYUN, LU JUNZHONG*, LIN NIN, ZHENG SHAOTONG
Department of Laboratory Medicine, the First Huaian Hospital Affiliated to Nanjing Medical
University, Huaian 223000, China
* Correspondence author.
Abstract
The crystal of 1, 2-bis (ethoxycarbonyl) hydrazine was studied by XRD, FTIR and Raman
spectroscopy. The ground-state geometries were optimized at B3LYP/6-31+G (d, p) level. These
vibrational wavenumbers were calculated at the same level. The antibacterial activities have been
screened for the compound against two bacteria: Staphylococcus aureus and Escherichia coli.
Keywords: Hydrazine; IR; Raman; DFT calculations; antibacterial activity
Introduction
Hydrazine is a chemical raw material with very extensive application. The subhydrazine contains two nucleophilic nitrogen and four replaceable hydrogens, therefore, it can
be synthesized into a number of hydrazine derivatives and important pharmaceutical
intermediate [1-6]. N, N- disubstituted hydrazine compounds is widely used in the chemical
industry, medicine, agriculture, aerospace and utility business and many other areas [7-9]. This
gave a great impetus to research the structure of the disubstituted hydrazine and to find the
potential valuable structure. The structure of 1, 2-bis (ethoxycarbonyl) hydrazine was studied
by XRD, FTIR and Raman spectroscopy. In addition, there is no theoretical study on 1, 2-bis
(ethoxycarbonyl) hydrazine before, in order to learn more about the compound, make full use
of it and even design important pharmaceuticals, a theoretical study of the vibration properties
of the compound by density-functional theory (DFT) was reported. The ground-state
geometries were optimized at B3LYP/6-31G (d, p) and the vibration wave numbers were
calculated at the same level. At the same time, the antibacterial activities have been screened
for the compound against two bacteria: Staphylococcus aureus and Escherichia coli.
Experiment
The structure of the compound was studied by X-ray single crystal diffraction by the
Siemens Smart 1000 instrument using Mo Kα as excitation source. Melting point was
measured in WRS-1B digital melting point instrument. Infrared spectra of the compound
dispersed in KBr pellet in the range of 400-4000cm-1 were recorded on an IR-Spectra One
(PE). The Raman spectra of powder sample were acquired on with Renishaw RM1000
spectrometer using Nd:YAG laser as excitation source (785 nm) with an output of about
200mW. The spectral regions were 400-3500cm-1.
Antibacterial activity: The bacteriostatic effect of the compound was measured using
single filter paper method against S. aureus (Staphylococcus aureus) and E. coli (Escherichia
coli). Protein in the beef extract agar medium, laboratory containers and direct diameter of
5.5mm filter paper were sterilized at 121°C for30 min. The medium was dished into the
containers as a flat after cooling to about 50°C, then were inoculated with the abovementioned two kinds of bacteria. The sterile filter paper was tiled in the bacterial culture
medium respectively, and was marked in a certain concentration of test fluid, cultured at 37°C
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LU JUNZHONG, FU QIYUN, LIN NIN, ZHENG SHAOTONG
for 24h, measure the inhibition zone diameters.
Calculations
The DFT calculations were employed with the Becke–Lee–Yang–Parr’s threeparameter hybrid functional (B3LYP) [10] method in this study. The basic set 6-31+G (d, p) [10]
was employed in the B3LYP calculations and the geometry optimizations were performed
without symmetry constrain. All calculations were performed with the Gaussian 03W
program suit [12]. The assignment of the calculated wave numbers is aided by the animation
option of Gauss View 3.0 [13] graphical interface for Gaussian programs, which gives a visual
presentation of the shape of the vibration modes. Note that the calculated spectra are all scaled
by 0.96, which is close to the recommended value by the NIST database (0.9614) [14] and is
comparable to other literature values [15].
Results and discussion
The cell diagram of 1, 2-bis (ethoxycarbonyl) hydrazine is shown in Fig. 1, it
crystallizes in monoclinic, space group C-2 with a=132.830(18)nm, b=0.87383(12)nm ,
c=0.76729(10)nm, α=90°, β=97.39°, γ=90°, Dc=1.325g·cm-3, Z=4, F(000)=376,
V=0.8832(2)nm3. The crystallographic data of 1, 2-bis (ethoxycarbonyl) hydrazine was
shown in Table 1.
Fig. 1. The cell diagram of 1,2-bis(ethoxycarbonyl)hydrazine.
Table 1. Crystallographic data of 1, 2-bis (ethoxycarbonyl) hydrazine.
Empirical formula
Crystal size/mm3
Formula weight
Temperature/K
Wavelength/nm
Crystal system
Space group
a/nm
b/nm
c/nm
α/(°)
β/(°)
γ/(°)
V/nm3
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C6 H12 N2 O4
0.30×0.20×0.10
176.18
292(2)
0.071073
Monoclinic
C2/c
1.32830(18)
0.87383(12)
0.76729(10)
90
97.39
90
0.8832(2)
Romanian Biotechnological Letters, Vol. 17, No. 5, 2012
The Structures of Hydrazine Crystal and Its Antibacterial Activities
Z
Dc/(g·cm-3)
F(000)
θ range for data collection
Index range
Reflections collected/unique
R(int)
Max. and min. transmission
R1,wR2[I>2σ(I)]
R1,wR2(all data)
GOF
Largest diff. Peak and
hole(e·nm-3)
4
1.325
376
2.80 to 27.98
-17<=h<=17, -11<=k<=11, 9<=l<=10
3727
0.0213
0.9890 and 0.9674
R1 = 0.0395, wR2 = 0.1103
R1 = 0.0434, wR2 = 0.1129
1.096
0.150 and -0.160
The schematic depiction of 1, 2-bis (ethoxycarbonyl) hydrazine and optimized
structure are shown in Fig. 2, and the optimized bond lengths, bond angles and dihedral
angles are shown in Table 2. From the table it can see that the calculated data are in
agreement with those determined by XRD. The bond length of N-N is 1.382 and 1.380 Å
respectively by calculation and XRD. The N-C bond is 1.385Å by calculation and 0.034Å
longer than that of by XRD. The C=O and C-O bond lengths are 1.217 and 1.346Å by
calculation, and they are 1.209 and 1.334Å by XRD.
Fig. 2. The optimized structure of 1,2-bis(ethoxycarbonyl)hydrazine.
Table 2. Selected bond lengths (Å), bond angles (◦), dihedral angles (◦) of 1, 2-bis (ethoxycarbonyl) hydrazine.
Parameters
B3LYP/6-31+G(d, p)
XRD
R(1,3)
1.382
1.380(18)
R(1,5)
1.385
1.351 (16)
R(5,7)
1.217
1.209 (14)
R (5,10)
1.346
1.334(13)
R(10,18)
1.453
1.454(15)
R(18,21)
1.520
1.496(19)
A(1,3,6)
123.5
120.0(9)
A(1,3, 4)
116.3
117.3(10)
A(3,6,8)
A(8,5,9)
A(6,9,11)
122.3
126.1
116.4
125.7(11)
125.4(11)
116.4(9)
A(9,11,14)
111.3
106.8(11)
D(1,5,10,18)
-177.6
-174.3(10)
Romanian Biotechnological Letters, Vol. 17, No. 5, 2012
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LU JUNZHONG, FU QIYUN, LIN NIN, ZHENG SHAOTONG
Table 3. Thermodynamic parameters of 1, 2-bis (ethoxycarbonyl) hydrazine. a Unscaled for 298K and 1 atm.
B3LYP/6-31+G(d,
Thermodynamic parametersa
p)
Zero-point vibrational energy (kJ·mol−1)
515.9
Sum of electronic and zero-point energy
-646.1
(kJ·mol−1)
551.9
Thermal correction to energy (kJ·mol−1)
510.7
Entropy (J·mol−1·K−1)
CV (J·mol−1·K−1)
197.4
-0.277
EHOMO (eV)
-0.001
ELUMO (eV)
Dipole moment (D)
1.067
The values of some thermodynamic parameters, of 1, 2-bis (ethoxycarbonyl)
hydrazine are listed in Table 3. The zero-point vibrational energy, the sum of electronic and
zero-point energy, the thermal correction to energy are 515.9, -646.1, 551.9 kJ·mol−1
respectively. The entropy, CV, EHOMO, ELUMO and the dipole moment are 510.7 J·mol−1·K−1,
197.4 J·mol−1·K−1, -0.277eV, -0.001eV, and 1.067D respectively. The FTIR and Raman
spectra of 1, 2-bis (ethoxycarbonyl) hydrazine were fully interpreted on the basis of the IR
and Raman spectra calculated at B3LYP/6-31+G(d, p) level, and the wave numbers are scaled
by the factor of 0.96. The intensity and assignments are listed in Table 4.
Table 4. The calculated wavenumbers (cm-1), FTIR and Raman intensity and assignments of the compound.
Abbreviation: ν– stretching, δ– deformation, ρw– wagging, ρb – bending, ρt – twisting, ρr – rocking, ρs –
scissoring, γ - torsion, oop – out of plane deformation, ip– in plane vibrations. as, asymmetric; s, symmetric. a
Scaling factor: B3LYP/6–31+G(d,p),0.96. b The absolute IR intensity in KM (mol)−1; Raman scattering activity
in °A4(amu)−1 from B3LYP/6–31+G(d,p) calculation.
Descriptions
νas(N–H)
νas(C-H)
νas(C-H)
νs(C-H)
νs (C-H)CH3
νas (C5–O7,C6–
ρ sC–H)
ρras (N-H)
ρrs (N-H)
ρbas (C–H)
ρbs (C–H)
γ(C–H)
ρr( C5-O7,C6νs(N–N)
ρt (C-H)
ρt (C-H)
νs(C18-O10,C11ρt (C-H)
γ(C–H)
7560
B3LYP/631+G(d,p)
3630
3151
3129
3087
3048
1793
1511
1485
1450
1436
1405
1332
1318
1251
1196
1117
1077
882
804
Calculated
B3LYP/631+G(d,p)a
3485
3025
3004
2964
2926
1721
1451
1426
1392
1379
1349
1279
1265
1201
1148
1072
1034
847
772
Observed
IR
intensityb
68
36.9
5.9
38.1
0.1
804.5
22.5
0.9
28.4
103.3
59
9.9
577
62.5
2.4
40
61.9
5
4.4
Raman
intensityb
43.22
12.9
78.5
166.5
295.26
1.58
3.5
10.19
0.73
1
0.19
10.91
1.38
0.35
1.22
0.61
4.34
1.75
0.79
IR
3415
3043
2988
2940
2912
1698
1448
1425
1388
1374
1336
1254
1240
1150
1067
1025
891(
780(
Raman
2980(s
2934(
1708(
1448(
1401(
1334(
1294(
1197(
1151(
1070(
1035(
871(vs
815(w
Romanian Biotechnological Letters, Vol. 17, No. 5, 2012
The Structures of Hydrazine Crystal and Its Antibacterial Activities
γ(C–H)
ρw(N-H)
ρrske
δske
ρsske
795
779
761
464
424
763
662
617
445
407
3.1
54.7
9.2
0.4
1.6
6.89
2.55
1.04
2.93
1.1
757(
671(
595(
453(
416(
795(w
670(w
598(m
466(w
431(w
Fig. 3 shows the observed FTIR and calculated IR spectra of 1, 2-bis (ethoxycarbonyl)
hydrazine at B3LYP/6-31+G (d, p) level. Fig. 4 shows the observed and calculated Raman
spectra of the compound at that level. These figures showed that the agreement between the
experimental and scaled theoretical frequencies is quite good in general. The weak band at
around 3415cm-1 is associated with the asymmetric stretching vibrations of N-H, and the
calculated band is at 3485cm-1. The bands near 3000cm-1 can be attributed to the C-H
stretching vibrations, and these can be seen from the theoretical results. The asymmetric
stretching vibration of C-H are at 3043, 2988cm-1 in FTIR spectra with media strong bands
and 2980cm-1 in Raman spectra with a strong band, while they are at 3025 and 3004 in
calculated spectra. The stretching vibration of C-H are at 2940, 2912cm-1 in FTIR spectra with
media strong and weak bands and 2934cm-1 in Raman spectra with a weak band, while they
are at 2964 and 2962cm-1 in calculated spectra. The very strong band at 1698cm-1 in FTIR
spectra and media strong band at 1708cm-1 in Raman spectra is attributed to the C5-O7 and
C6-C8 stretching vibration, and the calculated band is at 1721cm-1.
Fig. 3. The FTIR and unscaled IR spectra of 1,2-bis(ethoxycarbonyl)hydrazine at B3LYP/6-31+G(d,p) level.
Romanian Biotechnological Letters, Vol. 17, No. 5, 2012
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LU JUNZHONG, FU QIYUN, LIN NIN, ZHENG SHAOTONG
Fig. 4. The Raman and unscaled Raman spectra of 1,2-bis(ethoxycarbonyl)hydrazine at B3LYP/6-31+G(d,p)
level.
The media strong bands both at 1448cm-1 in FTIR spectra and Raman spectra are
attributed to the scissors vibration of C-H which was found at 1451cm-1 in calculated spectra.
The bands of asymmetric and symmetric rocking of N-H are at 1485 and 1450cm-1
respectively of the calculation, and these bands are observed at 1425 and 1388cm-1 in FTIR
spectra and 1401cm-1 in Raman spectra as weak bands. The weak band at 1374cm-1 in FTIR
spectra is attributed to the asymmetric bending of C-H, and the band of C-H symmetric
bending is observed at 1336cm-1 in FTIR spectra and 1334cm-1 in Raman spectra. The very
strong band at 1254cm-1 in FTIR spectra and weak band at 1294cm-1 in Raman spectra are
corresponding to the torsion of C-H, and the mode is computed at 1279cm-1. The very strong
band at 1240cm-1 in FTIR spectra and 1265cm-1 in the calculation spectra are attributed to the
rocking of C5-C7 and C6-C8. The weak band at 1197cm-1 in Raman spectra are attributed to
the N-N stretching, and it is found at 1201cm-1 in the calculation spectra. The torsion of C-H
is observed at 1150, 1607, 891cm-1 in FTIR spectra and 1150, 1070, 871cm-1 in Raman
spectra, and it is computed at 1148, 1072, 847cm-1. The bands at 1607 and 847cm-1 are very
strong. The media strong band at 1025cm-1 in FTIR spectra and 1035cm-1 in Raman spectra is
attributed to the symmetric stretching of C18-O10 and C11-O9 which calculated at 1034cm-1.
The media strong bands at 780, 757cm-1 in FTIR spectra and weak bands at 815, 795cm-1 in
Raman spectra are corresponding to the torsion of C-H, which are calculated at 772 and
763cm-1. The out of plane wagging of N-H is computed at 662cm-1 and is observed at 671cm-1
with a media strong band in FTIR spectra and 670cm-1 with a weak band in Raman spectra.
The media strong bands at 595cm-1 in FTIR spectra and 598cm-1 in Raman spectra are
attributed to the rocking of the skeleton, and it is calculated at 761cm-1. The weak bands at
453cm-1 in FTIR spectra and 466cm-1 in Raman spectra are attributed to the deformation of
the skeleton, and it is calculated at 445cm-1. The scissoring of the skeleton is calculated at
407cm-1 and is observed at 416cm-1 in FTIR spectra and 431cm-1 in Raman spectra.
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Romanian Biotechnological Letters, Vol. 17, No. 5, 2012
The Structures of Hydrazine Crystal and Its Antibacterial Activities
The results of antibacterial activity were shown in Table 5. It shows that the compound
has certain antibacterial activity against the two kinds of strains. The antibacterial activity of S.
aureus and E. coli increased as the concentration increasing. The diameter inhibition zone of E.
coli at the concentration of 1.5 mmol·L-1 is 10.1 mm, and it is 0.8 mm bigger than that of S.
aureus. So the antibacterial activity of the compound against E. coli is higher than S. aureus.
Table 5. Antibacterial activities of 1, 2-bis (ethoxycarbonyl) hydrazine.
Concentration(mmol·L
-1
)
0.5
1.0
1.5
Diameter inhibition
zone/mm
S. aureus
E. coli
8.1
8.3
8.8
9.4
9.3
10.1
Conclusions
The structure of 1, 2-bis (ethoxycarbonyl) hydrazine crystal was verified by XRD,
FTIR and Raman spectroscopy. The ground-state geometries were optimized at B3LYP/6-31G
(d, p) level. The vibrational wave numbers were calculated at the same level. It was found that
the calculated spectra are in agreement with the experimental data. The antibacterial activities
have been screened for the compound against two bacteria: Staphylococcus aureus and
Escherichia coli, resulting that the antibacterial activity of the compound against E. coli is
higher than that against S. aureus.
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