Content uploaded by Mahendra Kumar Trivedi
Author content
All content in this area was uploaded by Mahendra Kumar Trivedi on Aug 10, 2016
Content may be subject to copyright.
Science Journal of Analytical Chemistry
2016; 4(4): 42-51
http://www.sciencepublishinggroup.com/j/sjac
doi: 10.11648/j.sjac.20160404.11
ISSN: 2376-8045 (Print); ISSN: 2376-8053 (Online)
Determination of Isotopic Abundance of 13C/12C or 2H/1H
and 18O/16O in Biofield Energy Treated
1-Chloro-3-Nitrobenzene (3-CNB) Using Gas
Chromatography-Mass Spectrometry
Mahendra Kumar Trivedi
1
, Alice Branton
1
, Dahryn Trivedi
1
, Gopal Nayak
1
, Parthasarathi Panda
2
,
Snehasis Jana
2, *
1
Trivedi Global Inc., Henderson, Nevada, USA
2
Trivedi Science Research Laboratory Pvt. Ltd.,
Bhopal, Madhya Pradesh, India
Email address:
publication@trivedisrl.com (S. Jana)
*
Corresponding author
To cite this article:
Mahendra Kumar Trivedi, Alice Branton, Dahryn Trivedi, Gopal Nayak, Parthasarathi Panda, Snehasis Jana. Determination of Isotopic
Abundance of
13
C/
12
C or
2
H/
1
H and
18
O/
16
O in Biofield Energy Treated 1-Chloro-3-Nitrobenzene (3-CNB) Using Gas Chromatography-Mass
Spectrometry. Science Journal of Analytical Chemistry. Vol. 4, No. 4, 2016, pp. 42-51. doi: 10.11648/j.sjac.20160404.11
Received: May 10, 2016; Accepted: June 25, 2016; Published: July 23, 2016
Abstract:
1-Chloro-3-nitrobenzene (3-CNB) is an aromatic halo-amine compound used as chemical intermediate for the
production of several fine chemicals like pharmaceuticals, dyes, agricultural chemicals, etc. The stable isotope ratio analysis has
drawn attention in numerous fields such as agricultural, food authenticity, biochemistry, etc. The objective of the current research
was to investigate the impact of the biofield energy treatment on the isotopic abundance ratios of P
M+1
/P
M
, P
M+2
/P
M
and P
M+3
/P
M
in
3-CNB using gas chromatography - mass spectrometry (GC-MS). The sample, 3-CNB was divided into two parts - one part was
denoted as control and another part was referred as biofield energy treated sample that was treated with biofield energy (The
Trivedi Effect
®
). T1, T2, T3, and T4 were represented to different time interval analysis of the biofield treated 3-CNB. The GC-
MS spectra of the both control and biofield treated 3-CNB indicated the presence of molecular ion peak [M
+
] at m/z 157
(calculated 156.99 for C
6
H
4
ClNO
2
) along with same pattern of fragmentation. The relative intensities of the parent molecule and
other fragmented ions of the biofield treated 3-CNB were improved as compared to the control 3-CNB. The percentage change of
the isotopic abundance ratio of P
M+1
/P
M
was significantly increased in the biofield treated 3-CNB at T1, T2 and T3 by 11.62,
18.50, and 29.82%, respectively with respect to the control 3-CNB. Accordingly, the isotopic abundance ratio of P
M+2
/P
M
in the
biofield treated 3-CNB at T2 and T3 was significantly improved by 15.22 and 35.09%, respectively as compared to the control
sample. The isotopic abundance ratios of P
M+1
/P
M
and P
M+2
/P
M
in the biofield treated 3-CNB at T1 and T4 were changed as
compared to the control sample. The percentage change of the isotopic abundance ratio of P
M+3
/P
M
was enhanced in the biofield
treated 3-CNB at T1, T2, T3, and T4 by 4.67, 18.69, 31.31 and 6.08%, respectively as compared to the control 3-CNB. The
isotopic abundance ratios of P
M+1
/P
M
, P
M+2
/P
M
and P
M+3
/P
M
in the biofield treated 3-CNB changed with the time. So, the biofield
energy treated 3-CNB might exhibit the altered isotope effects such as altered physicochemical and thermal properties, binding
energy, and the rate of the chemical reaction as compared to the control sample. The biofield energy treated 3-CNB might assist in
designing for the synthesis of pharmaceuticals, agricultural chemicals, dyes, corrosion inhibitors and other several useful
industrial chemicals.
Keywords:
Biofield Energy Treatment, The Trivedi Effect
®
, 1-Chloro-3-Nitrobenzene,
Gas Chromatography - Mass Spectrometry, Isotopic Abundance Ratio, Isotope Effects, Kinetic Isotope Effect
43 Mahendra Kumar Trivedi et al.: Determination of Isotopic Abundance of
13
C/
12
C or
2
H/
1
H and
18
O/
16
O in Biofield
Energy Treated 1-Chloro-3-Nitrobenzene (3-CNB) Using Gas Chromatography-Mass Spectrometry
1. Introduction
Chloronitrobenzenes (CNBs) are aromatic halo-amines
and basically derivatives of monochlorobenzenes containing
nitro group in different positions with respect to the chloro
group. 1-Chloro-3-nitrobenzene or also commonly known as
3-chloronitrobenzene (3-CNB) as shown in the Figure 1 is
one of the isomeric forms of chloronitrobenzene. It is a pale
yellow crystalline solid having a molecular formula
C
6
H
4
ClNO
2
and molecular weight of 157.55.
Chloronitrobenzenes are widely used in the pharmaceutical
and chemical industries as an intermediates for the
production of pharmaceuticals, corrosion inhibitors, azo and
sulfur dyes, herbicides, pigments, agricultural chemicals,
rubber chemicals, photo chemicals, insecticides, and gasoline
additives [1-6]. 3-Chloroaniline (Orange GC Base), a dye
intermediate can be produced by reduction of 3-CNB.
Pentachloronitrobenzene which is used as fungicide can be
prepared by the exhaustive chlorination of 3-CNB [4-6].
Figure 1. Structure of 1-chloro-3-nitrobenzene (3-CNB).
Analysis of natural abundance variations in the stable
isotopes include
2
H,
13
C,
15
N,
18
O,
34
S,
37
Cl, etc. is a potential
method for the measurement of the flow of materials and
energy both within and among organisms. This is known as
Stable Isotope Ratio Analysis (SIRA). This method is
universally applied in agricultural, food authenticity,
biochemistry, metabolism, medical research, environmental
pollution, archaeology, etc. [7-10]. Isotope effects i.e. tiny
differences in physical and chemical properties of the
molecule are the resultant for the variation in isotopic
abundance ratio between isotopic forms of the molecule.
Isotope effects have an important role in thermal motion,
molecular spectra, chemical reactions (reaction rate and bond
strength), physicochemical properties, chemical equilibria,
etc. [11-15]. SIRA can also be used for the determination of
the pharmacokinetic profile or mode of action of a drug
substance, bioavailability of the drug products, the release
profile for the drug delivery systems and also used for the
assessment in relation to patient-specific drug treatment [8].
Among of the other technique like infrared spectroscopy,
nuclear magnetic resonance spectroscopy, and neutron
activation analysis, mass spectrometry (MS) technique such
as GC-MS is widely used for isotope ratio measurement at
low micromolar concentration levels with sufficient
precision. But when the molecules have molar isotope
enrichments at below 0.1%, specialized instruments, such as
isotope ratio mass spectrometer (IRMS), multiple-collector
inductively coupled plasma mass spectrometry are usually
used for the measurement of the ratio of natural isotopic
abundances in the molecule [8, 9, 14, 16]. Literature reported
that the peak height (i.e. relative intensity) in the mass
spectra is directly proportional to the relative isotopic
abundance of the sample [17-21].
Biofield is a dynamic electromagnetic field existing in
surrounds of the human body that carries information for
regulating the organism. Literature demonstrated that healing
practitioner has the capability to harness the energy from the
earth or environment, the “universal energy field” and can be
transmitted the biofield energy into any living or non-living
object (s) around the Globe in a useful way. This process is
known as biofield energy treatment [22, 23]. Mr. Trivedi is
one of the distinguished healing practitioners and has the
astonishingly ability to transform the characteristic properties
of several organic compounds [24-26], pharmaceuticals [27,
28], nutraceuticals [29], metals and ceramic in materials
science [30, 31], culture medium [32, 33] and improve the
overall productivity of crops [34, 35] as well as to modulate
the efficacy of the various living cells [36-38]. Literature
demonstrated that biofield energy treatment has the
remarkable capability for alteration of the isotopic abundance
ratio in the organic compounds [39-42]. Spectroscopic and
thermal analysis of 3-CNB inferred that the physicochemical,
structural and thermal properties of 3-CNB, such as
crystallite size, vaporization temperature and thermal
stability were significantly changed due to the biofield
energy treatment. Finally, it was suggested that these altered
properties might affect the reaction kinetics when it is used as
intermediate [6]. Hence, it is hypothesized that alteration of
the physicochemical, structural and thermal properties of
biofield treated 3-CNB might have a correlation with the
changes on the isotopic abundance ratio in biofield treated 3-
CNB. So, isotopic abundance ratio analysis of the both
control and biofield treated 3-CNB using GC-MS was
performed to investigate the influence of the biofield energy
treatment on the isotopic abundance ratios of P
M+1
/P
M
,
P
M+2
/P
M
and P
M+3
/P
M
in 3-CNB.
2. Materials and Methods
2.1. Chemicals and Reagents
3-CNB was procured from Loba Chemie Pvt. Ltd., India.
All the other chemicals used in this experiment were
analytical grade purchased from local vendors.
2.2. Biofield Energy Treatment
The sample 3-CNB was divided into two parts: one was
referred as control where no treatment was provided. The
other part of the sample which denoted as biofield energy
treated sample was handed over to Mr. Trivedi for the
biofield energy treatment in a sealed condition. The biofield
energy treatment was provided by Mr. Trivedi (also known as
The Trivedi Effect
®
) through his unique energy transmission
process to the test product in a sealed pack under laboratory
conditions for 5 minutes without touching the sample. After
Science Journal of Analytical Chemistry 2016; 4(4): 42-51 44
treatment, control and the biofield treated samples were
preserved at standard laboratory condition and analyzed by
GC-MS. The biofield treated 3-CNB was characterized in
different time intervals denoted as T1, T2, T3, and T4 in
order to understand the impact of the biofield energy
treatment on isotopic abundance ratio with respect to the
time.
2.3. Gas Chromatograph - Mass Spectrometry (GC-MS)
GC-MS analysis was performed on Perkin Elmer/Auto
system XL with Turbo mass, USA. The GC-MS was
conducted on a silica capillary column furnished with a
quadrupole detector with pre-filter. The mass spectrometer
was worked in an electron ionization (EI) positive/negative,
and chemical ionization mode at the electron ionization
energy of 70 eV. Mass range: 10-650 Daltons (amu),
stability: ± 0.1 m/z mass accuracy over 48 hours. The
analytes were identified by retention time and by a
comparison of the mass spectra of identified substances with
references [42].
2.4. Method for the Calculation of Isotopic Abundance
Ratio from the GC-MS Spectra
The isotopic abundances of the elements are basically
categorized into three types: A elements having only one
natural isotope in appreciable abundance; A + 1 elements
(For e.g. C, N and H) containing two isotopes – one isotope
is one nominal mass unit heavier than the most abundant
isotope, and A + 2 elements (For e.g. O, Cl, S, Si, and Br)
having an isotope that has two mass unit heavier than the
most abundant isotope [12, 20, 43]. The natural abundance of
each isotope can be predicted from the comparison of the
height of the isotope peak with respect to the base peak, i.e.
relative intensity in the mass spectra. The values of the
natural isotopic abundance of some elements are obtained
from several kind of literature and presented in the Table 1
[8, 12, 13, 43, 44].
Based on the findings from the literature [12, 13, 18-21],
the following method was used for calculating the isotopic
abundance ratio in the current study:
Table 1. The isotopic composition (i.e. the natural isotopic abundance) of the elements.
Element Symbol Mass % Natural Abundance A+1 Factor A+2 Factor
Hydrogen
1
H 1 99.9885
2
H
2 0.0115 0.015n
H
Carbon
12
C 12 98.892
13
C
13 1.108 1.1n
C
Oxygen
16
O 16 99.762
17
O 17 0.038 0.04n
O
18
O 18 0.200 0.20n
O
Nitrogen
14
N 14 99.60
15
N 15 0.40 0.40n
N
Chlorine
35
Cl 35 75.78
37
Cl 37 24.22 32.50n
Cl
A represents element, n represents the number of the element (i.e. C, H. O, N, etc.)
P
M
stands for the relative peak intensity of the parent
molecular ion [M
+
] expressed in percentage. In other way, it
indicates the probability to have A elements (for e.g.
12
C,
1
H,
16
O,
14
N, etc.) contributions to the mass of the parent
molecular ion [M
+
].
P
M+1
represents the relative peak intensity of the isotopic
molecular ion [(M+1)
+
] expressed in percentage
= (no. of
13
C x 1.1%) + (no. of
15
N x 0.40%) + (no. of
2
H x
0.015%) + (no. of
17
O x 0.04%)
i.e. the probability to have A + 1 elements (for e.g.
13
C,
2
H,
15
N, etc.) contributions to the mass of the isotopic molecular
ion [(M+1)
+
]
P
M+2
represents the relative peak intensity of the isotopic
molecular ion [(M+2)
+
] expressed in the percentage
= (no. of
18
O x 0.20%) + (no. of
37
Cl x 32.50%)
i.e. the probability to have A + 2 elements (for e.g.
18
O,
37
Cl,
34
S, etc.) contributions to the mass of isotopic molecular
ion [(M+2)
+
]
P
M+3
represents the relative peak intensity of the isotopic
molecular ion [(M+3)
+
] expressed in the percentage
i.e. the probability to have the different possible
combinations of
18
O and
37
Cl with
13
C,
2
H and
15
N
contributions to the mass of isotopic molecular ion [(M+3)
+
]
Isotopic abundance ratio for A + 1 elements = P
M + 1
/P
M
Similarly, isotopic abundance ratio for A + 2 elements =
P
M+2
/P
M
Percentage (%) change in isotopic abundance ratio =
[(IAR
Treated
– IAR
Control
)/ IAR
Control
) x 100],
Where, IAR
Treated
= isotopic abundance ratio in the treated
sample and IAR
Control
= isotopic abundance ratio in the
control sample.
3. Results and Discussion
3.1. GC-MS Analysis
The GC-MS spectra of the control and biofield treated 3-
CNB are presented in the Figures 2-4. The GC-MS spectrum
of the control 3-CNB (Figure 2) exhibited the presence of
molecular ion peak [M
+
] at m/z 157 (calculated 156.99 for
C
6
H
4
ClNO
2
) along with four major fragmented peaks in
lower m/z region at the retention time (R
t
) of 11.61 min. This
fragmentation pattern of CNB was well matched with the
literature [45]. The fragmented peaks at m/z 111, 99, 75 and
50 might be due to C
6
H
4
Cl
+
, C
6
H
11
O, C
6
H
3+
, and C
4
H
2
4+
ions,
respectively as shown in Figure 2.
45 Mahendra Kumar Trivedi et al.: Determination of Isotopic Abundance of
13
C/
12
C or
2
H/
1
H and
18
O/
16
O in Biofield
Energy Treated 1-Chloro-3-Nitrobenzene (3-CNB) Using Gas Chromatography-Mass Spectrometry
Figure 2. GC-MS spectrum and possible fragmentation of the control sample of 1-chloro-3-nitrobenzene (3-CNB).
The GC-MS spectra of the biofield treated 3-CNB at T1,
T2, T3, and T4 as shown in Figures 3 and 4 exhibited
molecular ion peak [M
+
] at m/z 157 at R
t
of 11.58, 11.64,
11.62, and 11.62 min, respectively. The biofield treated 3-
CNB showed similar R
t
and the same pattern of
fragmentation as observed in the control sample. The relative
peak intensities of the parent molecule and its major
fragmented ions of the control and biofield treated 3-CNB
are presented in the Table 2. It clearly indicated that the
fragmented ion peak at m/z 111 was due to chlorobenzenes
ion (C
6
H
4
Cl)
+
, which showed 100% relative intensity (base
peak). Table 2 also displayed that the relative intensities of
the parent molecule at m/z 157 and other fragmented ions at
m/z 99, 75, and 50 of the biofield treated 3-CNB were
significantly changed as compared with the control 3-CNB.
Figure 3. GC-MS spectra of the biofield energy treated 1-chloro-3-nitrobenzene (3-CNB) at T1 and T2.
Science Journal of Analytical Chemistry 2016; 4(4): 42-51 46
Table 2. Relative intensities of the corresponding m/z of the parent molecule (3-CNB) and its fragmented ions.
m/z
Relative intensity of the peak (%)
Control 3- CNB Biofield Energy Treated 3-CNB
T1 T2 T3 T4
157 46.63 41.93 87.48 93.79 49.89
111 100 100 100 100 100
99 24.88 24.38 60.73 51.90 25.38
75 87.91 99.15 90.48 92.01 82.49
50 32.32 43.55 70.22 65.89 35.31
T1, T2, T3, and T4: Biofield energy treated sample analyzed at different time intervals.
Figure 4. GC-MS spectra of the biofield energy treated samples of 1-chloro-3-nitrobenzene (3-CNB) at T3 and T4.
3.2. Analysis of Isotopic Abundance Ratio
3-CNB has the molecular formula of C
6
H
4
ClNO
2
and the
molecular ion [M
+
] peak for the control 3-CNB showed
46.63% relative intensity. P
M+1
and P
M+2
can be calculated
theoretically according to the method described in the
materials and method (section 2.4). The theoretical
calculation for P
M+1
is provided as follows:
P (
13
C) = [(6 x 1.1%) x 46.63% (the actual size of the M
+
peak)] / 100% = 3.08%
P (
2
H) = [(4 x 0.015%) x 46.63%] / 100%= 0.03%
P (
15
N) = [(1 x 0.40%) x 46.63%] / 100%= 0.19%
P (
17
O) = [(2 x 0.04%) x 46.63%] / 100% = 0.04%
P
M+1
i.e.
13
C,
2
H,
15
N, and
17
O contributions from
(C
6
H
4
ClNO
2
)
+
to m/z 158 = 3.34%
From the above calculation, it has been found that
13
C has
major contribution to m/z 169.
In the similar approach, P
M+2
can be calculated as follow:
P (
18
O) = [(2 x 0.20%) x 46.63%] / 100% = 0.19%
P (
37
Cl) = [(1 x 32.50%) x 46.63%] / 100% = 15.15%
So, P
M+2
i.e.
18
O and
37
Cl contributions from
(C
6
H
4
ClNO
2
)
+
to m/z 159 = 15.34%.
P
M
, P
M+1
, P
M+2
for the control and biofield energy treated
3-CNB at m/z 157, 158 and 159, respectively were
accomplished from the observed relative peak intensities of
[M
+
], [(M+1)
+
], and [(M+2)
+
] peaks in the GC-MS spectra,
respectively and are presented in the Table 3.
47 Mahendra Kumar Trivedi et al.: Determination of Isotopic Abundance of
13
C/
12
C or
2
H/
1
H and
18
O/
16
O in Biofield
Energy Treated 1-Chloro-3-Nitrobenzene (3-CNB) Using Gas Chromatography-Mass Spectrometry
Table 3. Isotopic abundance analysis result of the control and biofield energy treated 1-chloro-3-nitrobenzene (3-CNB).
Parameter Control 3-CNB
Biofield Energy Treated 3-CNB
T1 T2 T3 T4
P
M
at m/z 157 (%) 46.63 41.93 87.48 93.79 49.89
P
M+1
at m/z 158 (%) 3.05 3.06 6.78 7.96 3.41
P
M+1
/P
M
0.0654 0.0730 0.0775 0.0849 0.0684
% Change of isotopic abundance ratio (P
M+1
/P
M
) 11.62 18.50 29.82 4.59
P
M+2
at m/z 159 (%) 15.11 13.62 32.66 41.05 16.42
P
M+2
/P
M
0.3240 0.3248 0.3733 0.4377 0.3291
% Change of isotopic abundance ratio (P
M+2
/P
M
) 0.25 15.22 35.09 1.57
P
M+3
at m/z 160 (%) 1.00 0.94 2.22 2.64 1.13
P
M+3
/P
M
0.0214 0.0224 0.0254 0.0281 0.0227
% Change of isotopic abundance ratio (P
M+3
/P
M
) 4.67 18.69 31.31 6.08
T1, T2, T3, and T4: Biofield energy treated sample analyzed at different time intervals; P
M
= the relative peak intensity of the parent molecular ion [M
+
]; P
M + 1
= the relative peak intensity of the isotopic molecular ion [(M+1)
+
]; P
M + 2
= the relative peak intensity of the isotopic molecular ion [(M+2)
+
]; P
M + 3
= the
relative peak intensity of the isotopic molecular ion [(M+3)
+
]; and M = mass of the parent molecule.
The experimental values as shown in the Table 3 are well
accorded with the calculated theoretical values and it
indicated that
13
C and
37
Cl might have major contributions
from (C
6
H
4
ClNO
2
)
+
to m/z 158 and 159, respectively. Beside
these, an intense peak at m/z 160 [(M+3)
+
] that can be
denoted as P
M+3
was found in the GC-MS spectra of the both
control and biofield treated 3-CNB. It is assumed that P
M+3
might be resultant of the different possible combinations of
18
O and
37
Cl with
13
C,
2
H and
15
N for e.g. P (
37
Cl
13
C),
P(
15
N
18
O), P(
18
O
13
C), etc. The percentage change of the
isotopic abundance ratios (P
M+1
/PM, P
M+2
/PM and P
M+3
/P
M
)
in the biofield treated 3-CNB with respect to the control 3-
CNB is shown in Table 3 and Figure 5. The isotopic
abundance ratio of P
M+1
/PM in the biofield treated 3-CNB at
T1, T2, T3 and T4 was increased by 11.62, 18.50, 29.82, and
4.59%, respectively with respect to the control 3-CNB.
Consequently, the percentage change of the isotopic
abundance ratio of P
M+2
/P
M
was enhanced in the biofield
treated 3-CNB at T1, T2, T3, and T4 by 0.25, 15.22, 35.09,
and 1.57%, respectively with respect to the control sample.
Similarly, the isotopic abundance ratio of P
M+3
/P
M
was
improved in the biofield treated 3-CNB at T1, T2, T3, and T4
by 4.67, 18.69, 31.31 and 6.08%, respectively as compared to
the control 3-CNB. Thus,
13
C,
2
H,
15
N, and
17
O contributions
from (C
6
H
4
ClNO
2
)
+
to m/z 158,
37
Cl and
18
O contributions
from (C
6
H
4
ClNO
2
)
+
to m/z 159 and the different possible
combinations of
18
O and
37
Cl with
13
C,
2
H and
15
N
contributions from (C
6
H
4
ClNO
2
)
+
to m/z 160 in the biofield
treated 3-CNB were significantly increased gradually with
respect to the time (T1 to T3) and was found to be highest at
T3 as shown in the Figure 5. Amazingly, when biofield
treated 3-CNB was kept for long time in the laboratory
condition i.e. T4, the isotopic abundance ratio in biofield
treated 3-CNB was decreased from T3. So, the biofield
energy treatment exhibited time dependent effect on the
isotopic abundance ratio in 3-CNB.
Figure 5. Percent change of the isotopic abundance ratios of P
M+1
/P
M
,
P
M+2
/P
M
and P
M+3
/P
M
in the biofield energy treated 1-chloro-3-nitrobenzene
(3-CNB) as compared to the control sample.
Neutrinos are the most possible carrier of the hidden mass
in the nature. These electrically neutral particles that are part
of all living systems blast through the space and can pass
through large distances in the matter without being affected
the electromagnetic force. Literature suggested that the
neutrinos coming from the Sun have a potential effect on the
isotopic composition of the materials through inducing the
fission reactions within a heavy nuclei (i.e. the
nucleosynthesis of various elements) [46-48]. Trillions of
neutrinos are at any time passing through the human body
without affecting it. The biofield energy can freely flow
between human and environment that leads to the endless
movement or matter of energy [49-51]. It has been reported
that biofield energy might have effect on the variations of
isotopic composition in water molecule [23]. It can be
postulated that Mr. Trivedi’s unique biofield energy treatment
might have the capability for introduction of the neutrino
fluence into the both of the living and nonliving substances
that might responsible for modifying the behavior at atomic
and molecular level. The neutrinos have the ability to interact
Science Journal of Analytical Chemistry 2016; 4(4): 42-51 48
with protons and neutrons in the nucleus that might be
responsible for the alteration of the neutron to proton ratio in
the nucleus. Based on this hypothesis, it is assumed that the
possible reason for the alteration of the isotopic abundance
ratios (P
M+1
/P
M,
P
M+2
/P
M
and P
M+3
/P
M
) in the biofield treated
3-CNB might be due to the intervention of a neutrino flux
through biofield energy treatment.
The energy of a compound comprises of the amount of the
electronic, vibration, rotational and translation energies. The
alteration of the isotopic abundance ratio i.e. isotopic
composition of the molecule does not disturb electronic,
translational, and rotational energies of the molecule, but
significantly changes the vibrational energy [14, 15]. The
relation between the vibrational energy and the reduced mass
(µ) for a diatomic molecule is expressed as below [14, 15]:
=ℎ
4
Where E
0
= the vibrational energy of a harmonic oscillator
at absolute zero or zero point energy
f = force constant
µ = reduced mass =
,
Where m
a
and m
b
are the masses of the constituent atoms.
The possible isotopic bond formation in the CNB molecule
and their effect on the vibrational energy of 3-CNB are
shown in the Table 4. From the Table 4, it has been found
that the alteration of the isotopic abundance ratio of
13
C/
12
C
for C-C, C-Cl, and C-N bonds has much more effect on the
vibrational energy of the molecule than changes in the
isotopic abundance ratio of
37
Cl/
35
Cl and
15
N/
14
N. Similarly,
the changes of the isotopic abundance ratios of
2
H/
1
H for C-
H and
15
N/
14
N, and for
18
O/
16
O for N-O bond have much
more effect on the vibrational energy of the molecule. The
isotope effect is principally due to the ground state
vibrational energies as shown in the Table 4.
Table 4. Possible isotopic bond and their effect in the vibrational energy in 1-chloro-3-nitrobenzene (3-CNB) molecule.
Entry No. Probable isotopic bond Isotope type Reduced mass (µ) Zero point vibrational energy (E
0
)
1
12
C-
12
C Lighter 6.00 Higher
2
13
C-
12
C Heavier 6.26 Smaller
3
1
H-
12
C Lighter 0.92 Higher
4
1
H-
13
C Heavier 0.93 Smaller
5
2
H-
12
C Heavier 1.04 Smaller
6
12
C-
35
Cl Lighter 8.94 Higher
7
13
C-
35
Cl
Heavier 9.48 Smaller
8
12
C-
37
Cl
Heavier 9.06 Smaller
9
12
C-
14
N Lighter 6.46 Higher
10
12
C-
15
N Heavier 6.67 Smaller
11
13
C-
14
N Heavier 6.74 Smaller
12
14
N-
16
O
Lighter 7.47 Higher
13
15
N-
16
O
Heavier 7.74 Smaller
14
14
N-
17
O
Heavier 7.68 Smaller
15
14
N-
18
O
Heavier 7.88 Smaller
The isotopic abundance ratio analysis in 3-CNB clearly
revealed that the isotopic abundance ratios of P
M+1
/P
M
,
P
M+2
/P
M
and P
M+3
/P
M
in the biofield treated 3-CNB were
higher than the control 3-CNB. Hence, biofield treated 3-
CNB might exhibit altered isotope effects such as lower
diffusion velocity, mobility, evaporation and reaction rate,
higher binding energy [11] with respect to the control
sample. Isotope effects have a positive role in the thermal
decomposition of the molecules [52-54]. So, the alteration
in the isotopic abundance ratio in the molecule might have
an effect on the thermal properties of the molecule. Thus,
biofield treated 3-CNB might have altered
physicochemical and thermal properties as well as
different reaction kinetic than control 3-CNB. Hence, the
current results concluded that the increased isotopic
abundance ratio in biofield energy treated 3-CNB might
be responsible for alteration in vaporization rate and
thermal stability of the biofield treated 3-CNB that was
well supported with our previous findings [15]. The
alteration in the isotopic abundance ratio of one of the
atoms in the reactants causes changes in the rate of a
chemical reaction that is known as kinetic isotope effect
(KIE). KIE is a very powerful technique to study the
reaction mechanism, to stabilize the transition state of the
rate-determining step of the reaction and for
understanding the enzymatic transition state and all
aspects of enzyme mechanisms that is helpful for
designing enzyme inhibitors [14, 15, 55, 56]. Thus,
biofield treated 3-CNB might have altered
physicochemical and thermal properties, different rate of
the reaction, selectivity and binding energy.
49 Mahendra Kumar Trivedi et al.: Determination of Isotopic Abundance of
13
C/
12
C or
2
H/
1
H and
18
O/
16
O in Biofield
Energy Treated 1-Chloro-3-Nitrobenzene (3-CNB) Using Gas Chromatography-Mass Spectrometry
4. Conclusions
The present study concluded that biofield energy treatment
had potential impact on the isotopic abundance ratios of
P
M+1
/P
M
, P
M+2
/P
M
, and P
M+3
/P
M
in 3-CNB that might lead to
alteration of the physicochemical and thermal properties. The
GC-MS spectra of the both control and biofield treated 3-
CNB specified the presence of molecular ion peak [M
+
] at
m/z 157 (calculated 156.99 for C
6
H
4
ClNO
2
) along with
nearly similar fragmentation pattern. In addition, the relative
intensities of the parent molecule and other fragmented ions
of the biofield treated 3-CNB were altered with respect to the
control 3-CNB. The isotopic abundance ratio of P
M+1
/P
M
in
the biofield treated 3-CNB at T1, T2, T3 and T4 was
increased by 11.62, 18.50, 29.82, and 4.59%, respectively
with respect to the control 3-CNB. Consequently, the
percentage change of the isotopic abundance ratio of P
M+2
/P
M
was increased in the biofield treated 3-CNB at T1, T2, T3,
and T4 by 0.25, 15.22, 35.09, and 1.57%, respectively with
respect to the control sample. Similarly, the percentage of the
isotopic abundance ratio of P
M+3
/P
M
was improved in the
biofield treated 3-CNB at T1, T2, T3, and T4 by 4.67, 18.69,
31.31 and 6.08%, respectively with respect to the control 3-
CNB. In brief,
13
C,
2
H,
15
N, and
17
O contributions from
(C
6
H
4
ClNO
2
)
+
to m/z 158,
37
Cl and
18
O contributions from
(C
6
H
4
ClNO
2
)
+
to m/z 159 and the different possible
combinations of
18
O and
37
Cl with
13
C,
2
H and
15
N
contributions from (C
6
H
4
ClNO
2
)
+
to m/z 160 in the biofield
treated 3-CNB were significantly increased particularly at T2
and T3 and was found that biofield energy treatment has time
dependent effect on it. The biofield energy treated 3-CNB
might display the different isotope effects due to the
increased isotopic abundance ratio with respect to the control
sample. Hence, the biofield treated 3-CNB might have the
altered physicochemical and thermal properties and the rate
of the chemical reaction as compared to the control sample.
The biofield energy treated 3-CNB might play an important
role in designing the synthesis of pharmaceuticals,
agricultural chemicals, dyes, corrosion inhibitors and other
several useful industrial chemicals.
Abbreviations
A: Element; 3-CNB: 1-Chloro-3-nitrobenzene; GC-MS:
Gas chromatography-mass spectrometry; KIE: Kinetic
isotope effect; M: Mass of the parent molecule; m/z: Mass-to-
charge ratio; n: Number of the element; P
M
: The relative peak
intensity of the parent molecular ion [M
+
]; P
M+1
: The relative
peak intensity of isotopic molecular ion [(M+1)
+
]); P
M+2
: The
relative peak intensity of isotopic molecular ion [(M+2)
+
]);
P
M+3
: The relative peak intensity of isotopic molecular ion
[(M+3)
+
]); R
t
: Retention time.
Acknowledgements
The authors would like to acknowledge the Sophisticated
Instrumentation Centre for Applied Research and Testing -
SICART, Gujarat, India for providing the instrumental
facility. We are very grateful for the support from Trivedi
Science, Trivedi Master Wellness and Trivedi Testimonials in
this research work.
References
[1] Campos CH, Jofre M, Torres CC, Pawelec B, Fierro JLG,
Reyes P (2014) Chemoselective hydrogenation of o-, p- and
m-chloronitrobenzene at ambient temperature on Au/Fe
2
O
3
catalysts Appl Catal A- Gen 482: 127-136.
[2] http://www.sbioinformatics.com/design_thesis/Nitrochloro_B
enzene/NCB_Properties&uses.pdf
[3] Shen JM, Chen ZL, Xu ZZ, Li XY, Xu BB, Qi F (2008)
Kinetics and mechanism of degradation of p-chloronitrobenzene
in water by ozonation. J Hazard Mater 152: 1325-1331.
[4] http://monographs.iarc.fr/ENG/Monographs/vol65/mono65-
7.pdf
[5] Elvers B (2014) Nitro compounds, aromatic. Ullmann's Fine
Chemicals. Wiley-VCH, Weinheim.
[6] Trivedi MK, Branton A, Trivedi D, Nayak G, Singh R, Jana S
(2015) Characterization of biofield energy treated 3-
chloronitrobenzene: Physical, thermal, and spectroscopic
studies. J Waste Resources 5: 183.
[7] Gannes LZ, Martinez del Rio C, Koch P (1998) Natural
abundance variations in stable isotopes and their potential uses
in animal physiological ecology. Comp Biochem Physiol A
Mol Intergr Physiol 119: 725-737.
[8] Schellekens RC, Stellaard F, Woerdenbag HJ, Frijlink HW,
Kosterink JG (2011) Applications of stable isotopes in clinical
pharmacology. Br J Clin Pharmacol 72: 879-897.
[9] Muccio Z, Jackson GP (2009) Isotope ratio mass
spectrometry. Analyst 134: 213-222.
[10] http://www.eolss.net/sample-chapters/c06/e6-104-01-00.pdf
[11] www-
naweb.iaea.org/napc/ih/documents/global_cycle/.../cht_i_03.pd
[12] Smith RM (2004) Understanding Mass Spectra: A Basic
Approach, Second Edition, John Wiley & Sons, Inc, ISBN 0-
471-42949-X.
[13] Jürgen H. Gross Mass Spectrometry: A Textbook; Springer:
Berlin, (2ndedn), 2004.
[14] Vanhaecke F, Kyser K (2012) Isotopic composition of the
elements In Isotopic Analysis: Fundamentals and applications
using ICP-MS, 1
st
Edn., Edited by Vanhaecke F, Degryse P,
Wiley-VCH GmbH & Co. KGaA, Weinheim.
[15] Asperger S (2003) Chemical Kinetics and Inorganic Reaction
Mechanisms Springer science + Business media, New York.
[16] Meier-Augenstein W (1999) Applied gas chromatography
coupled to isotope ratio mass spectrometry. J Chromatogr A
842: 351-371.
[17] http://chemwiki.ucdavis.edu/Core/Analytical_Chemistry/Instr
umental_Analysis/Mass_Spectrometry/Mass_Spectrometry%3
A_Isotope_Effects
Science Journal of Analytical Chemistry 2016; 4(4): 42-51 50
[18] Raymond E. March, John F. J. Todd Practical Aspects of
Trapped Ion Mass Spectrometry, Volume IV: Theory and
instrumentation (2010) CRC press, Taylor and Francis Group,
Boca Raton.
[19] Skoog DA, Holler FJ, Crouch SR (2007) Principles of
instrument analysis, 6
th
Edn, Thomson Brooks/Cole, Australia.
[20] http://www.sepscience.com/Information/Archive/MS-
Solutions/254-/MS-Solutions-5-The-Role-of-Isotope-Peak-
Intensities-Obtained-Using-Mass-Spectrometry-in-
Determining-an-Elemental-Composition-Part-1
[21] https://www.unido.org/fileadmin/user_media/Services/Environ
mental_Management/Stockholm_Convention/POPs/DLWKSP
_Part2.pdf
[22] Rubik B (2002) The biofield hypothesis: Its biophysical basis
and role in medicine. J Altern Complement Med 8: 703-717.
[23] Klupenger MR (2015) An investigation into the effect of
human intention on the weather, Energy Medicine University,
Sausalito, California, USA.
[24] Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, Jana
S (2015) Impact of biofield treatment on spectroscopic and
physicochemical properties of p-nitroaniline. Insights in
Analytical Electrochemistry 1: 1-8.
[25] Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, Jana
S (2015) Physical, thermal and spectroscopical
characterization of biofield treated triphenylmethane: An
impact of biofield treatment. J Chromatogr Sep Tech 6: 292.
[26] Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, Jana
S (2015) Physicochemical and spectroscopic characteristics of
biofield treated p-chlorobenzophenone. American Journal of
Physical Chemistry 4: 48-57.
[27] Trivedi MK, Patil S, Shettigar H, Singh R, Jana S (2015) An
impact of biofield treatment on spectroscopic characterization
of pharmaceutical compounds. Mod Chem Appl 3: 159.
[28] Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, Jana
S (2015) Spectroscopic characterization of disulfiram and
nicotinic acid after biofield treatment. J Anal Bioanal Tech 6:
265.
[29] Trivedi MK, Tallapragada RM, Branton A, Trivedi D, Nayak
G, Latiyal O, Jana S (2015) Physical, atomic and thermal
properties of biofield treated lithium powder. J Adv Chem Eng
5: 136.
[30] Trivedi MK, Nayak G, Patil S, Tallapragada RM, Latiyal O
(2015) Impact of biofield treatment on physical, structural and
spectral properties of antimony sulfide. Ind Eng Manage 4:
165.
[31] Trivedi MK, Nayak G, Patil S, Tallapragada RM, Latiyal O
(2015) Studies of the atomic and crystalline characteristics of
ceramic oxide nano powders after bio field treatment. Ind Eng
Manage 4: 161.
[32] Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, Jana
S (2015) Effect of biofield treatment on physical, thermal, and
spectral properties of SFRE 199-1 mammalian cell culture
medium. Advances in Biochemistry 3: 77-85.
[33] Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, Jana
S (2015) Physicochemical and spectroscopic properties of
biofield energy treated protose. American Journal of
Biomedical and Life Sciences 3: 104-110.
[34] Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC,
Jana S (2015) Morphological characterization, quality, yield
and DNA fingerprinting of biofield energy treated alphonso
mango (Mangifera indica L.). Journal of Food and Nutrition
Sciences 3: 245-250.
[35] Trivedi MK, Branton A, Trivedi D, Nayak G, Gangwar M,
Jana S (2015) Agronomic characteristics, growth analysis, and
yield response of biofield treated mustard, cowpea, horse
gram, and groundnuts. International Journal of Genetics and
Genomics 3: 74-80.
[36] Trivedi MK, Patil S, Shettigar H, Gangwar M, Jana S (2015)
In Vitro evaluation of biofield treatment on cancer biomarkers
involved in endometrial and prostate cancer cell lines. J
Cancer Sci Ther 7: 253-257.
[37] Trivedi MK, Patil S, Shettigar H, Mondal SC, Jana S (2015)
An impact of biofield treatment: Antimycobacterial
susceptibility potential using BACTEC 460/MGIT-TB
system. Mycobact Dis 5: 189.
[38] Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC,
Jana S (2015) Antibiogram of biofield-treated Shigella boydii:
Global burden of infections. Science Journal of Clinical
Medicine 4: 121-126.
[39] Trivedi MK, Branton A, Trivedi D, Nayak G, Saikia G, Jana S
(2015) Quantitative determination of isotopic abundance ratio
of
13
C,
2
H, and
18
O in biofield energy treated ortho and meta
toluic acid isomers. American Journal of Applied Chemistry
3: 217-223.
[40] Trivedi MK, Branton A, Trivedi D, Nayak G, Saikia G, Jana S
(2015) Characterization of physico-chemical and
spectroscopic properties of biofield energy treated 4-
bromoacetophenone. American Journal of Physical Chemistry
4: 30-37.
[41] Trivedi MK, Branton A, Trivedi D, Nayak G, Saikia G, Jana S
(2015) Thermal, spectroscopic and chromatographic
characterization of biofield energy treated benzophenone.
Science Journal of Analytical Chemistry 3: 109-114.
[42] Trivedi MK, Branton A, Trivedi D, Nayak G, Saikia G, Jana
S (2015) Investigation of isotopic abundance ratio of
biofield treated phenol derivatives using gas
chromatography-mass spectrometry. J Chromatograph
Separat Techniq S6: 003.
[43] http://www.ionsource.com/Card/Mass/mass.htm
[44] Meija J, Coplen TB, Berglund M, Brand WA, De Bievre P,
Groning M, Holden NE, Irrgeher J, Loss RD, Walczyk T,
Prohaska T (2016) Isotopic compositions of the elements 2013
(IUPAC technical Report). Pure Appl Chem 88: 293-306.
[45] http://webbook.nist.gov/cgi/cbook.cgi?ID=C121733&Mask=2
00#Mass-Spec
[46] Clayton D (2003) Handbook of Isotopes in the Cosmos:
Hydrogen to Gallium, Cambridge University Press, New
York.
[47] Balantekin AB (2013) Neutrinos and rare isotopes Journal of
Physics: Conference Series 445 012022.
[48] Domogatskii GV, Nadezhin DK (1978) Neutrino production
of bypassed isotopes, and the possible role of neutrinos in
nucleosynthesis. Astron Zh 55: 516-530.
51 Mahendra Kumar Trivedi et al.: Determination of Isotopic Abundance of
13
C/
12
C or
2
H/
1
H and
18
O/
16
O in Biofield
Energy Treated 1-Chloro-3-Nitrobenzene (3-CNB) Using Gas Chromatography-Mass Spectrometry
[49] Rogers, M (1986) "Science of Unitary Human Beings." In V.
M. Malinski (ed.) Explorations of Martha Rogers' Science of
Unitary Human Beings. Norwark: Appleton Century Crofts.
[50] Jenkins L (2011) Healing in the present moment. ISBN 978-1-
257-77329-9.
[51] de Climont J (2016) The worldwide list of dissident scientists,
ISBN 978-2-9024-2517-4.
[52] Carr Jr. RW, Walters WD (1966) The hydrogen isotope effect
in the thermal decomposition of cyclobutane. J Am Chem Soc
88: 884-887.
[53] Lomas JS, Thorne MP (1982) Structure and isotope effects
upon the thermal decomposition of carbamates of highly
congested tertiary alcohols. J Chem Soc, Perkin Trans 2 221-
226.
[54] Makhatadze GI, Clore GM, Gronenborn AM (1995) Solvent
isotope effect and protein stability. Nat Struct Biol 2: 852 -
855.
[55] Schramm VL (1998) Enzymatic transition states and transition
state analog design. Annu Rev Biochem 67: 693-720.
[56] Cleland WW (2003) The use of isotope effects to determine
enzyme mechanisms. J Biol Chem 278: 51975-51984.
