Evaluation of the Impact of the Trivedi Effect® -Energy of Consciousness on the Structure and Isotopic Abundance Ratio of Magnesium Gluconate Using LC-MS and NMR Spectroscopy

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American Journal of Biomedical and Life Sciences
2017; 5(1): 6-15
http://www.sciencepublishinggroup.com/j/ajbls
doi: 10.11648/j.ajbls.20170501.12
ISSN: 2330-8818 (Print); ISSN: 2330-880X (Online)
Evaluation of the Impact of the Trivedi Effect
®
-Energy of
Consciousness on the Structure and Isotopic Abundance
Ratio of Magnesium Gluconate Using LC-MS and NMR
Spectroscopy
Mahendra Kumar Trivedi
1
, Alice Branton
1
, Dahryn Trivedi
1
, Gopal Nayak
1
, Michael Peter Ellis
1
,
James Jeffery Peoples
1
, James Joseph Meuer
1
, Johanne Dodon
1
, John Lawrence Griffin
1
,
John Suzuki
1
, Joseph Michael Foty
1
, Judy Weber
1
, Julia Grace McCammon
1
, Karen Brynes Allen
1
,
Kathryn Regina Sweas
1
, Lezley Jo-Anne Wright
1
, Lisa A. Knoll
1
, Madeline E. Michaels
1
,
Margaret Kweya Wahl
1
, Mark E. Stutheit
1
, Michelle Barnard
1
, Muriel Mae Ranger
1
,
Paromvong Sinbandhit
1
, V. J. Kris Elig
1
, Kalyan Kumar Sethi
2
, 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@trivedieffect.com (S. Jana)
*
Corresponding author
To cite this article:
Mahendra Kumar Trivedi, Alice Branton, Dahryn Trivedi, Gopal Nayak, Michael Peter Ellis, James Jeffery Peoples, James Joseph Meuer,
Johanne Dodon, John Lawrence Griffin, John Suzuki, Joseph Michael Foty, Judy Weber, Julia Grace McCammon, Karen Brynes Allen,
Kathryn Regina Sweas, Lezley Jo-Anne Wright, Lisa A. Knoll, Madeline E. Michaels, Margaret Kweya Wahl, Mark E. Stutheit, Michelle
Barnard, Muriel Mae Ranger, Paromvong Sinbandhit, V. J. Kris Elig, Kalyan Kumar Sethi, Parthasarathi Panda, and Snehasis Jana.
Evaluation of the Impact of the Trivedi Effect
®
-Energy of Consciousness on the Structure and Isotopic Abundance Ratio of Magnesium
Gluconate Using LC-MS and NMR Spectroscopy. American Journal of Biomedical and Life Sciences. Vol. 5, No. 1, 2017, pp. 6-15.
doi: 10.11648/j.ajbls.20170501.12
Received: February 7, 2017; Accepted: February 18, 2017; Published: March 9, 2017
Abstract:
Magnesium gluconate is a classical pharmaceutical/nutraceutical compound used as a magnesium ion source for
the prevention and treatment of hypomagnesemia. The present study was aimed to investigate the effect of The Trivedi Effect
®
- Energy of Consciousness Healing Treatment (Biofield Energy Healing Treatment) on magnesium gluconate for the change in
the structural properties and isotopic abundance ratio (P
M+1
/P
M
and P
M+2
/P
M
) using LC-MS and NMR spectroscopy.
Magnesium gluconate was divided into two parts – one part was control, and another part was treated with The Trivedi Effect
®
- Biofield Energy Healing Treatment remotely by twenty renowned Biofield Energy Healers and defined as The Trivedi
Effect
®
Treated sample. The LC-MS analysis of both the control and treated samples indicated the presence of mass of the
protonated magnesium gluconate at m/z 415 at the retention time of 1.52 min and fragmentation pattern of the both sample
were almost similar. The relative peak intensities of the fragment ions were significantly changed in the treated sample
compared with the control sample. The proton and carbon signals for CH, CH
2
and CO groups in the proton and carbon NMR
spectra were observed almost similar for the control and the treated samples. The percentage change in the isotopic abundance
ratio of P
M+1
/P
M
(
2
H/
1
H or
13
C/
12
C or
17
O/
16
O or
25
Mg/
24
Mg) was significantly decreased in the treated sample by 17.51%
compared with the control sample. Consequently, the isotopic abundance ratio of P
M+2
/P
M
(
18
O/
16
O or
26
Mg/
24
Mg) in the treated
sample was significantly increased by 79.44% compared to the control sample. Briefly,
13
C,
2
H,
17
O, and
25
Mg contributions
from (C
12
H
23
MgO
14
)
+
to m/z 416;
18
O and
26
Mg contributions from (C
12
H
23
MgO
14
)
+
to m/z 417 in treated sample were
significantly altered compared with the control sample. Thus, The Trivedi Effect
®
Treated magnesium gluconate might be
supportive to design the novel potent enzyme inhibitors using its kinetic isotope effects. Consequently, The Trivedi Effect
®
Treated magnesium gluconate would be valuable for designing better pharmaceutical and/or nutraceutical formulations through
American Journal of Biomedical and Life Sciences 2017; 5(1): 6-15 7
its changed physicochemical and thermal properties, which might be providing better therapeutic response against various
diseases such as diabetes mellitus, allergy, aging, inflammatory diseases, immunological disorders, and other chronic
infections.
Keywords:
Magnesium Gluconate, Biofield Energy Healing Treatment, Consciousness Energy Healing Treatment,
Biofield Energy Healers, The Trivedi Effect
®
, LC-MS, NMR, Isotopic Abundance Ratio, Isotope Effect
1. Introduction
Magnesium gluconate is a classical pharmaceutical/
nutraceutical compound used for the source of magnesium
ion, a major and essential mineral in the human body [1].
Magnesium is a vital element for more than 300 enzymes,
DNA and RNA synthesis, reproduction and protein synthesis
as well as a vital coherent controller of glycolysis and the
Krebs cycle [2, 3]. Hypomagnesemia may cause several
diseases and disorders [4-7]. Magnesium gluconate is found
to be the most physiologically acceptable antioxidant and
showed the highest level of bioavailability of magnesium
among the commercially available magnesium salts such as
chloride, sulfate, citrate, lactate, aspartate, etc. [8-10]. Thus,
magnesium gluconate has the important application for the
prevention and treatment of diabetes mellitus, allergies,
cardiovascular diseases, septic shock, inflammatory diseases,
immunological disorders, arrhythmias, acute myocardial
infarction, gestational hypertension, preeclampsia, eclampsia,
Alzheimer's disease, cancer, and oxidative stress induced
ischemia/reperfusion injury [4-7, 9-11]. It can be used as
neuroprotective [12], an oral tocolytic agent [13], and also in
a skin-tightening cosmetic composition [14]. Subsequently,
magnesium gluconate was considered as one of the
components in a novel proprietary herbomineral formulation,
which is designed as nutraceutical for the source of
magnesium ion.
Since ancient times, many different cultures, religions and
systems of belief have recognized a living force that preserves
and inhabits every living organism. This force is the source of
‘life’ and has been called various names, such as prana by the
Hindus, qi or chi by the Chinese, and ki by the Japanese. This
is believed to co-relate with the soul, spirit and mind. This
hypothetical vital force has been scientifically evaluated and is
now considered the Bioenergetics Field. The Biofield Energy
is a dynamic electromagnetic field surrounding the human
body, resulting from the continuous emission of low-level
light, heat, and acoustical energy from the body. Biofield
Energy is infinite, para-dimensional and can freely flow
between the human and environment [15]. F. Sances et al.
reported that Biofield Energy can be transmitted into any
living organism (s) or nonliving object (s) around the globe
with scientifically measurable effect through the intentional
mental energies by specific energy healers [16]. The object or
recipient always receives the energy from the ionosphere of the
earth, the “universal energy field” and responds in a useful
way. This process is known as The Trivedi Effect
®
- Biofield
Energy Healing Treatment [17, 18]. Biofield (Putative Energy
Field) based Energy Therapies are used worldwide to promote
health and healing. The National Center of Complementary
and Integrative Health (NCCIH) has recognized and accepted
Biofield Energy Healing as a Complementary and Alternative
Medicine (CAM) health care approach in addition to other
therapies, medicines and practices such as natural products,
deep breathing, yoga, Tai Chi, Qi Gong,
chiropractic/osteopathic manipulation, meditation, massage,
special diets, homeopathy, progressive relaxation, guided
imagery, acupressure, acupuncture, relaxation techniques,
hypnotherapy, healing touch, movement therapy, pilates,
rolfing structural integration, mindfulness, Ayurvedic
medicine, traditional Chinese herbs and medicines,
naturopathy, essential oils, aromatherapy, Reiki, cranial sacral
therapy and applied prayer (as is common in all religions, like
Christianity, Hinduism, Buddhism and Judaism) [19]. Biofield
Energy Treatment (The Trivedi Effect
®
) has been drawn
attention more in the recent times for its scientifically
measurable capability to transform the characteristic properties
of a wide varieties living and non-living substances such as
microbes [20, 21], cancer cells [22], plants [23, 24], animals
[25], medium [26, 27], materials [28, 29], pharmaceuticals [30,
31], nutraceuticals [32, 33], organic compounds [34, 35]. The
scientific study indicated that Biofield Energy Healing
Treatment (The Trivedi Effect
®
) might be an alternate method
for increasing or decreasing the natural isotopic abundance
ratio of the substances [36-39]. The stable isotope ratio
analysis has the broad application in several scientific fields
for understanding the isotope effects resulting from the
difference of the isotopic composition of the molecule [40, 41].
Conventional mass spectrometry (MS) techniques such as
liquid chromatography-mass spectrometry (LC-MS), gas
chromatography-mass spectrometry (GC-MS) are extensively
used for isotope ratio analysis with sufficient precision [42].
Hence, LC-MS and NMR (Nuclear Magnetic Resonance)
methods were applied in this research work to depict the
structural properties of the Biofield Energy Treated and
untreated magnesium gluconate. Consequently, the authors
sought to explore the impact of The Trivedi Effect
®
(Consciousness Energy Healing Treatment) on the isotopic
abundance ratios of P
M+1
/P
M
and P
M+2
/P
M
in magnesium
gluconate using LC-MS based isotopic abundance ratio
analysis in both the Biofield Energy Treated and untreated
samples.
2. Materials and Methods
2.1. Chemicals and Reagents
Magnesium gluconate hydrate was purchased from Tokyo
8 Mahendra Kumar Trivedi et al.: Evaluation of the Impact of the Trivedi Effect
®
-Energy of Consciousness on the
Structure and Isotopic Abundance Ratio of Magnesium Gluconate Using LC-MS and NMR Spectroscopy
Chemical Industry Co., Ltd. (TCI), Japan. All other
chemicals used in the experiment were of analytical grade
available in India.
2.2. Energy of Consciousness Healing Treatment Strategies
Magnesium gluconate was one of the components of the
new proprietary herbomineral formulation, which was
developed by our research team and was used per se as the
test compound for the current study. The test compound was
divided into two parts, one part of the test compound did not
receive any sort of treatment and was defined as the untreated
or control magnesium gluconate sample. The second part of
the test compound was subjected to The Trivedi Effect
®
-
Energy of Consciousness Healing Treatment (Biofield
Energy Healing Treatment) by a group of twenty renowned
Biofield Energy Healers remotely and was denoted as The
Trivedi Effect
®
- Biofield Energy treated sample. Thirteen
Biofield Energy Healers were remotely located in the U. S.
A., five were located in Canada, and two were located in
Australia, while the test compound was located in the
research laboratory of GVK Biosciences Pvt. Ltd.,
Hyderabad, India. The Trivedi Effect
®
- Energy of
Consciousness Healing Treatment (Biofield Energy Healing
Treatment) was provided for 5 minutes through the Healers
Unique Energy Transmission process remotely to the test
compound, which was kept under laboratory conditions.
None of the Biofield Energy Healers in this study visited the
laboratory in person, nor had any contact with the
compounds. Similarly, the control compound was subjected
to “sham” healer for 5 minutes, under the same laboratory
conditions. The sham healer did not have any knowledge
about the Biofield Energy Treatment. After that, The Trivedi
Effect
®
treated and untreated samples were kept in similar
sealed conditions and characterized thoroughly by LC-MS
and NMR spectroscopy.
2.3. Liquid Chromatography Mass Spectrometry (LC-MS)
Analysis
Liquid chromatography was performed using The Waters
®
ACQUITY UPLC, Milford, MA, USA equipped with a
binary pump (The Waters
®
BSM HPLC pump), autosampler,
column heater and a photo-diode array (PDA) detector. The
column used for the study was a reversed phase Acquity
BEH shield RP C18 (150 X 3.0 mm, 2.5 µm). The column
temperature was kept constant at 40°C. The mobile phase
was 2mM ammonium acetate in water as mobile phase A and
acetonitrile as mobile phase B. Chromatographic separation
was achieved with following gradient program: 0 min
5%B; 1 min – 5%B; 15 min - 97%B; 20 min – 97%B; 21 min
5%B; 25 min 5%B. The flow rate was at a constant flow
rate of 0.4 mL/min. The control and treated samples were
dissolved in a mixture of water and methanol (60:40 v/v) to
prepare a 1 mg/mL stock solution. An aliquot of 2 µL of the
stock solution was used for analysis by LC-ESI-MS and the
total run time was 25 min. Mass spectrometric analysis was
accompanied on a Triple Quad (Waters Quattro Premier XE,
USA) mass spectrometer equipped with an electrospray
ionization (ESI) source with the following parameters:
electrospray capillary voltage 3.5 kV; source temperature
100°C; desolvation temperature 350°C; cone voltage 30. V;
desolvation gas flow 1000 L/h and cone gas flow 60 L/h.
Nitrogen was used in the electrospray ionization source. The
multiplier voltage was set at 650 V. LC-MS was taken in
positive ionization mode and with the full scan (m/z 50-
1400). The total ion chromatogram, % peak area and mass
spectrum of the individual peak (appeared in LC) were
recorded.
2.4. Isotopic Abundance Ratio Analysis
The relative intensity of the peak in the mass spectra is
directly proportional to the relative isotopic abundance of the
molecule and the isotopic abundance ratio analysis was
followed by the scientific literature reported [36-39] method
described as below:
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 A elements having only one
natural isotope in appreciable abundance (for e. g.
12
C,
1
H,
16
O,
24
Mg, 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%) + (no. of
25
Mg x 12.66%)
i.e. the probability to A + 1 elements having an isotope that
has one mass unit heavier than the most abundant isotope (for
e. g.
13
C,
2
H,
17
O,
25
Mg, 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
26
Mg x 13.94%)
i.e. the probability to have A + 2 elements having an
isotope that has two mass unit heavier than the most
abundant isotope (for e. g.
18
O,
26
Mg, etc.) contributions to the
mass of isotopic molecular ion [(M+2)
+
].
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 1.1 n
C
Oxygen
16
O 16 99.762
17
O 17 0.038 0.04 n
O
18
O 18 0.200 0.20 n
O
Magnesium
24
Mg 24 78.99
25
Mg 25 10.00 12.66 n
26
Mg 26 11.01 13.94 n
A represents element, n represents the number of the
element (i.e. C, H. O, Mg, etc.)
The value of the natural isotopic abundance of the
elements used here for the theoretical calculation are
American Journal of Biomedical and Life Sciences 2017; 5(1): 6-15 9
achieved from the scientific literature and are presented in
the Table 1 [43, 44].
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 Biofield
Energy Treated sample and IAR
Control
= isotopic abundance
ratio in the control sample.
2.5. Nuclear Magnetic Resonance (NMR) Analysis
1
H NMR spectra were recorded in a 400 MHZ VARIAN
FT-NMR spectrometer at room temperature. Data refer to
solutions in D
2
O with the residual solvent protons as internal
references.
1
H NMR multiplicities were designated as singlet
(s), doublet (d), triplet (t), multiplet (m), and broad (br).
13
C
NMR spectra were measured at 100 MHz on a VARIAN FT-
NMR spectrometer at room temperature. Chemical shifts (δ)
were in parts per million (ppm) relative to the solvent’s
residual proton chemical shift (D
2
O, δ = 4.65 ppm) and
solvent’s residual carbon chemical shift (D
2
O, δ = 0 ppm).
3. Results and Discussion
3.1. Liquid Chromatography-Mass Spectrometry (LC-MS)
Analysis
The liquid chromatograms of both the control and treated
magnesium gluconate (Figure 1) showed a sharp and narrow
peak at the retention time (R
t
) of 1.52 min. This finding
indicated that the polarity/affinity of the Trivedi Effect
®
-
Biofield Energy Treated sample remained same compared
with the control sample.
The ESI-MS spectra of both the control and treated
magnesium gluconate at R
t
of 1.52 min (Figure 2) exhibited
the presence of the mass of magnesium gluconate at m/z 415
[M + H]
+
(calcd for C
12
H
23
MgO
14
+
, 415).
Figure 1. Liquid chromatograms of the control and Biofield Energy Treated magnesium gluconate.
Figure 2. The ESI-MS spectra of the control and Biofield Energy Treated magnesium gluconate.
The typical fragment ion peaks in the lower m/z region of
the protonated magnesium gluconate ion [M]
+
(m/z 415) were
observed in both the control and treated samples at m/z 402
[M H
2
O + 6H]
+
(calcd for C
12
H
26
MgO
13
4+
, 402), 381 [M
2H
2
O + 3H]
+
(calcd for C
12
H
21
MgO
12
+
, 381), 361 [M – 3H
2
O
+ H]
+
(calcd for C
12
H
17
MgO
11
+
, 361), and 343 [M 4H
2
O +
H]
+
(calcd for C
12
H
15
MgO
10
+
, 343). The fragment ion peaks
at m/z 320, 307, 279, 225, 206, 183, 165, 142, 135, and 100
which correspond to the following molecular formula
C
10
H
16
MgO
10
+
, C
9
H
15
MgO
10
+
, C
9
H
3
MgO
9
2+
, C
8
H
9
MgO
6
+
,
10 Mahendra Kumar Trivedi et al.: Evaluation of the Impact of the Trivedi Effect
®
-Energy of Consciousness on the
Structure and Isotopic Abundance Ratio of Magnesium Gluconate Using LC-MS and NMR Spectroscopy
C
8
H
6
MgO
5
2+
, C
6
H
7
MgO
5
+
, C
5
H
9
O
6
+
, C
5
H
2
O
5
2+
, C
5
H
11
O
4
+
,
and C
4
H
4
O
3
2+
, respectively as shown in the Figure 3 were
observed in the ESI-MS spectra of both the control and
treated samples (Figure 2).
Figure 3. Proposed fragmentation pathway of magnesium gluconate.
Besides, the fragment ion peak at m/z 255, 212, 123 and
115 corresponding to the molecular formula C
9
H
11
MgO
7
+
,
C
7
H
8
MgO
6
2+
, C
4
H
11
O
4
+
, and C
5
H
7
O
3
+
, respectively were only
found in the control sample. Consequently, the fragment ion
peaks at m/z 254, 213, 124, and 114 corresponding to the
molecular formula C
9
H
10
MgO
7
2+
, C
7
H
9
MgO
6
+
, C
4
H
12
O
4
2+
,
and C
5
H
6
O
3
2+
were observed in the ESI-MS spectrum of the
treated sample. The ESI-MS spectra of the control and
treated samples (Figure 2) exhibited almost similar type
fragmentation pattern. The fragment ion peak at m/z 165
corresponding to C
5
H
9
O
6
+
showed 100% relative peak
intensity in the ESI-MS spectrum of the control sample,
while the highest intense peak was observed in the ESI-MS
spectrum of the treated sample at m/z 114 corresponding to
C
5
H
6
O
3
2+
(Figure 2). The relative peak intensities of the other
ion peaks in the treated sample were significantly altered
compared with the control sample.
3.2. Isotopic Abundance Ratio Analysis
The molecular formula of magnesium gluconate is
C
12
H
22
MgO
14
. But, it existed as a protonated molecular ion at
m/z 415 (C
12
H
23
MgO
14
+
) in the ESI-MS spectrum of the
control sample that exhibited 34.73% relative intensity. The
theoretical calculation of P
M+1
and P
M+2
for the protonated
magnesium gluconate in the control sample was presented as
below:
P (
13
C) = [(12 x 1.1%) x 34.73% (the actual size of the M
+
peak)] / 100% = 4.58%
P (
2
H) = [(23 x 0.015%) x 34.73%] / 100%= 0.12%
P (
17
O) = [(14 x 0.04%) x 34.73%] / 100% = 0.19%
P (
25
Mg) = [(1 x 12.66%) x 34.73%] / 100% = 4.40%
P
M+1
i. e.
13
C,
2
H,
17
O, and
25
Mg contributions from
(C
12
H
23
MgO
14
)
+
to m/z 416 = 9.29%
From the above calculation, it has been found that
13
C and
25
Mg have major contribution to m/z 416.
In the similar way, P
M+2
can be calculated as follow:
P (
18
O) = [(14 x 0.20%) x 34.73%] / 100% = 0.97%
P (
26
Mg) = [(1 x 13.94%) x 34.73%] / 100% = 4.84%
So, P
M+2
i.e.
18
O and
26
Mg contributions from
(C
12
H
23
MgO
14
)
+
to m/z 417 = 5.81%.
The calculated isotopic abundance ratios of P
M+1
/P
M
and
P
M+2
/P
M
were near to the observed value (Table 2). The above
calculation indicated that
13
C and
25
Mg have the major
contributions from magnesium gluconate to the isotopic peaks.
LC-MS spectra of the control and treated samples
indicated the presence of the mass for the protonated
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