Physicochemical and Spectroscopic Characterization of Biofield Treated Butylated Hydroxytoluene

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Volume 1 • Issue 1 • 1000101J Food Ind Microbiol, an open access journal
Open Access
Research Article
Trivedi et al., J Food Ind Microbiol 2015, 1:1
Journal of
Food & Industrial Microbiology
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*Corresonding author: Jana S, Trivedi Science Research Laboratory Pvt. Ltd.,
Hall-A, Chinar Mega Mall, Chinar Fortune City, Hoshangabad Rd., Bhopal- 462026,
Madhya Pradesh, India, Tel: +91-755-6660006; E-mail: publication@trivedieffect.com
Received September 23, 2015; Accepted September 29, 2015; Published
October 09, 2015
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Singh R, et al. (2015)
Physicochemical and Spectroscopic Characterization of Bioeld Treated Butylated
Hydroxytoluene. J Food Ind Microbiol 1: 101.
Copyright: © 2015 Trivedi MK, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited.
Abstract
The antioxidants play an important role in the preservation of foods and the management of oxidative stress related
diseases by acting on reactive oxygen species and free radicals. However, their use in high temperature processed food and
pharmaceuticals are limited due to its low thermal stability. The objective of the study was to use the bioeld energy treatment on
butylated hydroxytoluene (BHT) i.e. antioxidant and analyse its impact on the physical, thermal, and spectral properties of BHT.
For the study, the sample was divided into two groups and termed as control and treated. The treated group was subjected to
bioeld energy treatment. The characterization of treated sample was done using X-ray diffraction (XRD), differential scanning
calorimetry (DSC), thermogravimetric analysis (TGA), Fourier transform infrared (FT-IR) and UV-visible (UV-Vis) spectroscopy.
The XRD results showed the alteration in lattice parameters, unit cell volume, and molecular weight along with 14.8% reduction in
the crystallite size of treated sample as compared to the control. The DSC analysis showed an increase in the latent heat of fusion
from 75.94 J/g (control) to 96.23 J/g in the treated BHT sample. The TGA analysis showed an increase in onset temperature of
decomposition (130°C→136°C) and maximum thermal decomposition temperature (152.39°C→158.42°C) in the treated sample
as compared to the control. Besides, the FT-IR analysis reported the shifting of aromatic C-H stretching peak towards higher
frequency (3068→3150 cm
-1
) and C=C stretching towards lower frequency (1603→1575 cm
-1
) as compared to the control sample.
Moreover, the UV spectrum also revealed the shifting of the peak at λ
max
247 nm (control) to 223 nm in the treated sample. The
overall results showed the impact of bioeld energy treatment on physical, thermal and spectral properties of BHT sample.
Physicochemical and Spectroscopic Characterization of Biofield Treated
Butylated Hydroxytoluene
Trivedi MK
1
, Branton A
1
, Trivedi D
1
, Nayak G
1
, Singh R
2
and Jana S
2
*
1
Trivedi Global Inc., 10624 S Eastern Avenue Suite A-969, Henderson, NV 89052, USA
2
Trivedi Science Research Laboratory Pvt. Ltd., Hall-A, Chinar Mega Mall, Chinar Fortune City, Hoshangabad Rd., Bhopal-462026, Madhya Pradesh, India
Keywords: Bioeld energy treatment; Butylated hydroxytoluene;
Reactive oxygen species; Complementary and alternative medicine;
ermogravimetric analysis
Introduction
In recent years, the studies on reactive oxygen species (ROS),
free radicals and antioxidants are generating medical revolution by
promising a good health and disease management [1]. e free radicals
and ROS can be developed inside the human body either through
the normal metabolic process or external sources such as pollutants,
industrial chemicals, and cigarette smoking, etc. [2]. An antioxidant is
a molecule that neutralises these free radicals by donating an electron
to them. ey prevent the oxidative reaction that is responsible for
various chronic degenerative diseases viz. cancer, cardiovascular and
neurodegenerative disorders, etc. [3]. Besides, in the pharmaceutical
industry, the safety, ecacy and stability of drug formulations are
aected by various physical factors like humidity, heat, and light
[4]. ese factors are responsible for several chemical reactions that
cause instability such as oxidation, decarboxylation, hydrolysis, and
photolysis, etc. [5]. e antioxidants are such excipients that can
enhance the shelf-life of the drug by reducing the problem due to
oxidation reactions. Butylated hydroxytoluene (BHT) is a monohydric
phenol derivative (Figure 1) that provides electrons or protons labile
to free radicals and interrupts the chain reaction thereby exerting its
antioxidant action [6]. It is used as antioxidant in foods containing
fats and oils as they are very susceptible to rancidity and oxidation
that destroy the soluble vitamins and fatty acids. BHT prevents this
rancidication by terminating the free radical chain reactions [7,8].
It helps in food preservation by preventing any change in avour and
slowing the rancidity and discoloration processes [9]. Moreover, it is
also added in personal care products to prevent oxidative rancidity
thereby disagreeable smell [10]. e antioxidants are also added in
several manufacturing processes to protect the raw material and end
product from deleterious eects of high temperature and pressure [11].
In food preservation process, the thermal stability of antioxidant is
very crucial. Several edible oils are used at higher temperature; hence
thermal stability of antioxidant is necessary to preserve the unsaturated
fatty acid from degradation [12]. Despite several uses, BHT is sensitive
to heat, light and humidity and is also known to possess some toxic and
ammable properties hence need special precautions during storage,
handling, and transportation [13,14]. Moreover, BHT is reported
to decompose at low temperature due to which it oers very less
protection to heated vegetable oils [15]. us, it is important to search
Figure 1: Chemical structure of butylated hydroxytoluene.
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Singh R, et al. (2015) Physicochemical and Spectroscopic Characterization of Bioeld Treated
Butylated Hydroxytoluene. J Food Ind Microbiol 1: 101.
Page 2 of 7
Volume 1 • Issue 1 • 1000101J Food Ind Microbiol, an open access journal
some alternate strategies, which could improve the stability of BHT by
altering their physical, thermal or structural and bonding properties.
Nowadays, bioeld energy treatment is known to alter
physicochemical properties of various organic and metallic compounds
[16-19]. e bioeld energy healing therapies are considered as
complementary and alternative medicine (CAM) by National Center
for Complementary and Alternative Medicine (NCCAM)/National
Institute of Health (NIH) and are based on alteration in putative
energy elds and consciousness [20,21]. e bioeld energy is related
to the energy associated with the human body that depends upon the
physiological and mental health of the human. is energy can be
exchanged with the environment through natural exchange process
[22]. A human has the ability to harness the energy from environment
or universe and can transmit in any living or non-living object(s)
around the Universe. e objects always receive the energy and
responding in a useful way, this process is known as bioeld energy
treatment. Moreover, the bioeld therapies are reported for the
reduction in pain, anxiety and tension [23]. Mr. Trivedi is well known
to possess the unique bioeld energy treatment (e Trivedi Eect
®
)
which is reported to alter the properties such as growth and yield of
plants in the eld of agriculture [24,25]. e eect was also reported on
phenotypic characters of microorganisms in the eld of microbiology
[26,27]. Hence, the present study was designed to analyse the impact
of bioeld energy treatment on various physicochemical properties of
BHT using XRD, DSC, TGA/DTG, FT-IR and UV-Vis spectroscopic
techniques.
Materials and Methods
Material procurement
Butylated hydroxytoluene (BHT) was procured from S D Fine
Chemicals Pvt. Ltd., India. Aer procurement, the BHT sample was
divided into two parts; coded as control and treated, and stored as per
manufacturer’s guidelines.
Treatment modality
e treated part was subjected to Mr. Trivedi’s bioeld energy
treatment. For this, the treated sample was handed over to Mr. Trivedi
in sealed pack for bioeld energy treatment under standard laboratory
conditions. Mr. Trivedi provided the treatment to the treated group
through his energy transmission process. e bioeld treated sample
was returned in the same sealed condition for further characterization
using XRD, DSC, TGA, FT-IR and UV-Vis spectroscopic techniques.
X-ray diraction (XRD) study
X-ray powder diractogram of control and treated samples were
obtained on Phillips, Holland PW 1710 X-ray diractometer system.
e X-ray generator was equipped with a copper anode with nickel
lter operating at 35 kV and 20 mA. e wavelength of radiation used
by the XRD system was 1.54056 Å. e data were collected from the 2θ
range of 10°-99.99° with a step size of 0.02° and a counting time of 0.5
seconds per step.
e crystallite (G) was calculated from the Scherrer equation with
the method based on the width of the diraction patterns obtained in
the X-ray reected crystalline region.
G = kλ/(bCosθ)
Where, k is the equipment constant (0.94), λ is the X-ray wavelength
(0.154 nm), b in radians is the full-width at half of the peak and θ the
corresponding Bragg angle. Other parameters viz. lattice parameter
and unit cell volume, were calculated using PowderX soware. Further,
these parameters were used to calculate the molecular weight and
density of the control and treated sample.
Percent change in crystallite size was calculated using the following
equation:
Percent change in crystallite size = [(G
t
-G
c
)/G
c
] ×100
Here, G
c
and G
t
denotes the crystallite size of control and treated
powder samples, respectively. Similarly the percent change in lattice
parameter, unit cell volume, molecular weight, and density was
calculated to analyse the impact of bioeld treatment on crystal
parameters of treated sample as compared to the control.
e molecular weight of atom was calculated using following
equation:
Molecular weight = number of electrons × weight of an electron
+ number of neutrons × weight of a neutron + number of protons ×
weight of a proton.
e weight of all atoms in a molecule was multiplied by the
Avogadro number (6.023 × 10
23
) to obtain the molecular weight in g/
Mol.
Dierential scanning calorimetry (DSC) study
DSC analysis of control and treated sample was carried out using
Perkin Elmer/Pyris-1. Each sample was accurately weighed and
hermetically sealed in aluminium pans and heated at a rate of 10°C/
min under air atmosphere (5 mL/min). e thermogram was collected
over the temperature range of 50°C to 250°C. An empty pan sealed with
cover pan was used as a reference sample. From DCS curve, the melting
temperature and latent heat of fusion were obtained.
e percent change in latent heat of fusion was obtained using
following equations to observe the dierence in thermal properties of
treated BHT sample as compared to the control:
Treated Control
Control
H
% change in lat
H
100ent heat of fusion
H
−∆


×
=
Where, ΔH
Control
and ΔH
Treated
denotes the latent heat of fusion of
control and treated samples, respectively.
ermogravimetric analysis/Derivative ermogravimetry
(TGA/DTG)
e eect of temperature on the stability of the control and treated
sample of BHT was analysed using Mettler Toledo simultaneous
thermogravimetric analyser (TGA/DTG). e samples were heated
from room temperature to 350°C with a heating rate of 5°C/min under
air atmosphere. From TGA/DTG curve, the onset temperature T
onset
(temperature at which sample start losing weight) and T
max
(maximum
thermal degradation temperature) were recorded.
Fourier transform-infrared (FT-IR) spectroscopic
characterization
For FT-IR characterization, the treated sample was divided into two
groups named T1 and T2. e samples were crushed into ne powder
for analysis. e powdered sample was mixed in spectroscopic grade
KBr in an agate mortar and pressed into pellets with a hydraulic press.
FT-IR spectra were recorded on Shimadzu’s Fourier transform infrared
spectrometer (Japan). FT-IR spectra are generated by the absorption of
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Singh R, et al. (2015) Physicochemical and Spectroscopic Characterization of Bioeld Treated
Butylated Hydroxytoluene. J Food Ind Microbiol 1: 101.
Page 3 of 7
Volume 1 • Issue 1 • 1000101J Food Ind Microbiol, an open access journal
electromagnetic radiation in the frequency range 4000-400 cm
-1
. With
the help of FT-IR analysis, the impact of bioeld treatment on bond
strength, rigidity and stability of BHT compound can be analysed [28].
UV-Vis spectroscopic analysis
For UV-Vis spectroscopic analysis, the treated sample was divided
into two groups, served as T1 and T2. e UV-Vis spectral analysis
was measured using Shimadzu UV-2400 PC series spectrophotometer.
e spectrum was recorded with 1 cm quartz cell having a slit width
of 2.0 nm over a wavelength range of 200-400 nm. With UV-Vis
spectroscopy, it is possible to investigate electron transfers between
orbitals or bands of atoms, ions and molecules from the ground state to
the rst excited state [29].
Results and Discussion
X-ray diraction (XRD)
e X-ray powder diractograms of control and treated samples
showing Bragg angle (2θ) on x-axis and intensity of the peaks on y-axis
are presented in Figure 2. e XRD diractograms showed a series of
sharp peaks in the regions of 10°<2θ>40°, which depicted that both
samples had high crystallinity and long range order of molecules.
e sharp peaks on the diractograms of the control and treated
sample conrm the crystalline nature of BHT [30]. e XRD pattern
indicated the orthorhombic crystal structure in the control and treated
BHT samples. e crystal structure parameters were computed using
PowderX soware such as lattice parameter, unit cell volume, density,
and molecular weight. e results are presented in Table 1. e data
showed that in the control sample, the lattice parameters were found
as a=15.46, b=10.37, and c=8.93 Å. While, in treated sample, the lattice
parameters were found as a=15.59, b=10.56 and c=8.71 Å. It showed
that the lattice parameters ‘a’ and ‘b’ were increased by 0.84% and
1.83%, respectively. However, the lattice parameter ‘c’ was decreased by
2.46% in the treated sample as compared to the control. Similarly, the
unit cell volume and molecular weight was slightly increased by 0.09%
and 0.08%, respectively; whereas, density was decreased by 0.10% in
the treated sample as compared to the control. e increase in unit
cell volume and change in lattice parameters indicated the presence of
internal strain in the treated BHT powder. Besides, the crystallite size
computed using Scherrer formula was found as 70.16 nm in control
and it was reduced to 59.77 nm in the treated BHT sample. It suggested
that crystallite size of the treated sample was signicantly reduced
by 14.81% as compared to the control. It is reported that the energy
produced by mechanical milling had reduced the crystallite size and
induced lattice strain in the crystal structure [31]. us, it is assumed
that bioeld energy treatment might induce the energy milling in BHT
sample, and that might be responsible for a decrease in crystallite size
of treated sample. Recently, our group reported that bioeld treatment
had reduced the crystallite size in magnesium powder [19]. Moreover,
it was reported that crystallite size and surface area are inversely related
to each other [32]. e BHT had a poor solubility prole in water that
limits its application in pharmaceutical preparations [7]. Hence, the
decreased crystallite size of treated BHT might result in increased
surface area, and that can play an important role in improving
solubility. erefore, the treated BHT sample may be used in food and
pharmaceutical industry with improved solubility prole.
DSC analysis
is technique is based on the principle that as the BHT sample
undergoes any phase transition (e.g. solid to liquid); the alteration was
observed in the amount of heat owing to the sample as compared to
the reference. e thermograms for control and treated sample of BHT
are presented in Figure 3 that showed the phase transition temperature
and the amount of heat involved in that process. e control sample
exhibited a sharp endothermic peak at 71.6°C, whereas the treated
sample showed a sharp peak at 72.25°C. e peaks are due to melting
of control and treated samples, and the sharpness of peaks conrms the
crystalline nature of BHT sample. e result suggests a slight change
in melting temperature of treated sample as compared to the control.
e thermograms also showed that the latent heat of fusion (ΔH) was
increased from 75.94 J/g (control) to 96.23 J/g in treated BHT sample. It
indicated that ΔH was signicantly increased by 26.72% in the treated
sample as compared to the control. It is presumed that bioeld energy
might increase the potential energy stored in the molecules of treated
BHT sample. Hence, the treated sample needs more energy in the
form of ΔH to undergo the process of melting. Previously, our group
reported that bioeld treatment has altered the latent heat of fusion in
indole, thymol and menthol compounds [17,33].
Control
Treated
Figure 2: XRD of control and treated samples of butylated hydroxytoluene.
Parameter Control Treated Percent
change
Lattice parameter
a (Å) 15.46 15.59 0.84
b (Å) 10.37 10.56 1.83
c (Å) 8.93 8.71 -2.46
Unit cell volume (×10
-23
cm
3
) 143.208 143.330 0.090
Density (w/cm
3
) 1.029 1.028 -0.10
Molecular weight (g/mol) 222.752 222.941 0.08
Crystallite size (nm) 70.16 59.77 -14.81
Table 1: X-ray diffraction analysis of control and treated butylated
hydroxytoluene.
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Singh R, et al. (2015) Physicochemical and Spectroscopic Characterization of Bioeld Treated
Butylated Hydroxytoluene. J Food Ind Microbiol 1: 101.
Page 4 of 7
Volume 1 • Issue 1 • 1000101J Food Ind Microbiol, an open access journal
TGA/DTG analysis
e TGA thermograms (Figure 4) of the control and treated samples
of BHT showed single step decomposition within the temperature
range of 100-200°C. From these thermograms, the information related
to degradation temperature and the weight of sample lost in that event
was collected. e control sample started to decompose around 130°C
(onset) and completed around 175°C (end set). However, the treated
sample started losing weight around 136°C (onset) and terminated at
185°C (end set). It indicated that onset temperature of decomposition
was increased in the treated sample as compared to the control. Besides,
DTG thermogram data showed that T
max
was found at 152.39°C in the
control sample, whereas, it was increased to 158.42°C in the treated
BHT sample. It indicated that T
max
was also increased in treated
sample as compared to the control. Furthermore, the increase in onset
temperature of decomposition and T
max
in the treated sample of BHT
with respect to the control sample may be correlated with the increase
in thermal stability of the treated sample aer bioeld treatment. e
data was also supported by DSC studies, which revealed that ΔH of
treated BHT sample increased as compared to the control sample. As
BHT is reported to decompose at higher temperature due to which
it oers very less protection to food ingredients. Moreover, BHT is
considered as a ammable material and increase in thermal stability
may be related to the decreased ammability of compound [13,34].
Hence, it was presumed that the bioeld energy treatment might
enhance the thermal stability of the treated BHT sample. e increase
in thermal stability may enhance its eectiveness in food ingredients at
high temperature as well as might assure its safe handling by decreasing
the ammability as compared to the control sample.
FT-IR spectroscopic analysis
Infrared (IR) spectroscopy is based on the vibrations of the atoms
in a molecule. e FT-IR spectra of control and treated (T1 and T2)
samples of BHT are shown in Figure 5 that has the wavenumber
(frequency) of IR rays on the horizontal axis and percent transmittance
on the vertical axis. e comparative values of IR peaks of the control
sample with treated (T1 and T2) samples are given in Table 2. e
major vibration peaks observed were as follows:
O-H vibrations: e O-H vibrations are sensitive to hydrogen
bonding; however the OH group present in the structure of BHT did
not show intermolecular hydrogen bonding due to steric shielding of
tert-butyl groups present in the structure. e non-bonded hydroxyl
groups in phenols showed absorption in 3700-3584 cm
-1
region [35]. In
the present study, the strong band observed at 3628 cm
-1
in control and
T2 sample and 3626 cm
-1
in T1 sample was assigned to OH stretching
mode of vibration. e strong band observed at 1151 cm
-1
in all three
samples (control, T1, and T2) was assigned to OH in-plane bending
vibration.
Aromatic C-H vibrations: e aromatic CH stretching vibration
was observed as a weak band at 3068 cm
-1
in control and T2 sample
whereas at 3150 cm
-1
in T2 sample. e in-plane CH bending vibrations
was observed at 1230 cm
-1
in all three samples (control, T1 and T2).
Methyl group vibrations: In the present study, the CH
3
asymmetric
vibration was observed at 2956 cm
-1
in the control sample and 2955 cm
-1
in both treated samples (T1 and T2). e CH
3
symmetric stretching
mode was observed at 2872 cm
-1
in all three samples i.e. control, T1,
and T2. Similarly, the bands observed at 1481 and 1431 cm
-1
in control
and T1 and 1481 and 1433 cm
-1
in T2 were assigned to the CH
3
bending
vibrations. e CH
3
rocking vibration was observed at 1026 cm
-1
in all
three samples (control, T1 and T2).
Tert-butyl group vibrations: In the FT-IR spectra, the strong,
sharp bands observed at 1361 and 1396 cm
-1
in the control sample and
1363 and 1396 cm
-1
in treated samples (T1 and T2) were assigned to
tert-butyl bending vibrations.
C-O vibrations: In the present study, the FT-IR band observed at
1213 cm
-1
in control and T1 sample and 1215 cm
-1
in T2 sample was
Control
Treated
Figure 3: DSC thermogram of control and treated samples of butylated
hydroxytoluene.
S. No. Functional group Wavenumber (cm
-1
)
Control T1 T2
1. O-H stretching 3628 3626 3628
2. C-H stretching (aromatic) 3068 3150 3068
3. C-H
3
asymmetric stretching 2956 2955 2955
4. C-H
3
symmetric stretching 2872 2872 2872
5. C=C stretching (aromatic) 1603 1575 1602
6. C-H
3
bending 1481
1431
1481
1431
1481
1431
7. Tert-butyl group 1396
1361
1396
1363
1396
1363
8. C-H bending (in plane) 1230 1230 1230
9. C-O stretching (C-OH) 1213 1213 1215
10. O-H bending (in plane) 1151 1151 1151
11. C-H
3
rocking 1026 1026 1026
12. C-C ring stretching 866
769
866
769
866
769
13. Aromatic ring bending
(out of plane)
580 580 580
T1 and T2 are treated samples
Table 2: Vibration modes observed in butylated hydroxytoluene.
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Singh R, et al. (2015) Physicochemical and Spectroscopic Characterization of Bioeld Treated
Butylated Hydroxytoluene. J Food Ind Microbiol 1: 101.
Page 5 of 7
Volume 1 • Issue 1 • 1000101J Food Ind Microbiol, an open access journal
assigned to C-O stretching mode of C-OH vibration.
C=C vibrations: e ring stretching vibration was observed at
1603 cm
-1
in control sample, whereas at 1575 and 1602 cm
-1
in T1 and
T2 samples, respectively. e bands observed at 769 and 866 cm
-1
were
assigned to C-C ring stretching mode. Moreover, the band at 580 cm
-1
was assigned to ring out-of-plane bending mode in all three samples
(control, T1, and T2).
e FT-IR spectrum of the control sample of BHT was well
supported by literature [35,36]. e FT-IR spectra of treated BHT
samples (T1 and T2) showed the similar pattern of IR absorption
peaks as control sample except C-H aromatic stretching peak and C=C
aromatic stretching peak in T1 sample. e C-H aromatic stretching
peak was shied to higher frequency (3068→3150 cm
-1
); whereas, C=C
aromatic stretching peak was shied to lower frequency (1603→1575
cm
-1
) as compared to the control sample.
When a molecule absorbs
infrared radiation, its chemical bonds vibrate due to which they can
stretch, contract or bend. It was already reported that the peak frequency
Control
Treated
Figure 4: TGA/DTG thermograms of control and treated samples of butylated hydroxytoluene.
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