Structural and Natural Isotopic Abundance Analysis of Magnesium Gluconate Treated with Energy of Consciousness (The Trivedi Effect®) Using LC-MS and NMR

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Chemical and Biomolecular Engineering
2017; 2(3): 124-134
http://www.sciencepublishinggroup.com/j/cbe
doi: 10.11648/j.cbe.20170203.11
Structural and Natural Isotopic Abundance Analysis of
Magnesium Gluconate Treated with Energy of
Consciousness (The Trivedi Effect
®
) Using LC-MS and NMR
Mahendra Kumar Trivedi
1
, Alice Branton
1
, Dahryn Trivedi
1
, Gopal Nayak
1
, Aileen Carol Lee
1
,
Aksana Hancharuk
1
, Carola Marina Sand
1
, Debra Jane Schnitzer
1
, Rudina Thanasi
1
,
Eileen Mary Meagher
1
, Faith Ann Pyka
1
, Gary Richard Gerber
1
, Johanna Catharina Stromsnas
1
,
Judith Marian Shapiro
1
, Laura Nelson Streicher
1
, Lorraine Marie Hachfeld
1
,
Matthew Charles Hornung
1
, Patricia M. Rowe
1
, Sally Jean Henderson
1
, Sheila Maureen Benson
1
,
Shirley Theresa Holmlund
1
, Stephen P. Salters
1
, Parthasarathi Panda
2
, Kalyan Kumar Sethi
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, Aileen Carol Lee, Aksana Hancharuk, Carola Marina Sand, Debra
Jane Schnitzer, Rudina Thanasi, Eileen Mary Meagher, Faith Ann Pyka, Gary Richard Gerber, Johanna Catharina Stromsnas, Judith Marian
Shapiro, Laura Nelson Streicher, Lorraine Marie Hachfeld, Matthew Charles Hornung, Patricia M. Rowe, Sally Jean Henderson, Sheila
Maureen Benson, Shirley Theresa Holmlund, Stephen P. Salters, Parthasarathi Panda, Kalyan Kumar Sethi, Snehasis Jana. Structural and
Natural Isotopic Abundance Analysis of Magnesium Gluconate Treated with Energy of Consciousness (The Trivedi Effect
®
) Using LC-MS
and NMR. Chemical and Biomolecular Engineering. Vol. 2, No. 3, 2017, pp. 124-134. doi: 10.11648/j.cbe.20170203.11
Received: January 31, 2017; Accepted: February 14, 2017; Published: March 9, 2017
Abstract: The current research article was aimed to investigate the impact of The Trivedi Effect
®
- Energy of Consciousness
Healing Treatment on the structural properties and isotopic abundance ratio (P
M+1
/P
M
and P
M+2
/P
M
) of magnesium gluconate
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
®
remotely by eighteen renowned Biofield Energy Healers and defined as the Trivedi
Effect
®
Treated sample. The liquid chromatogram of the control sample showed two peaks at R
t
of 1.81 and 2.06 min, whereas
the Trivedi Effect
®
Treated sample displayed these peaks at R
t
of 1.79 and 2.03 min. The ESI-MS spectra of the control and the
Trivedi Effect
®
Treated samples revealed the presence of the mass for magnesium gluconate ion in two forms at m/z 447
(adduct form with methanol) and 415 (protonated ion) in positive ionization mode. But, it showed the mass for the gluconate
ion at m/z 195 in the negative ionization mode. The fragmentation pattern of magnesium gluconate in the treated sample was
notably altered as compared to the control sample. The proton and carbon signals for CH, CH
2
and CO groups in the proton
and carbon NMR spectra were found almost similar for the control and the treated samples. The isotopic abundance ratio
analysis revealed that 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) in two magnesium
gluconate ion forms at m/z 447 and 415 in treated sample was significantly decreased by 59.82% and 55.44%, respectively
compared with the control sample. The percentage change in the isotopic abundance ratio of P
M+2
/P
M
(
18
O/
16
O or
26
Mg/
24
Mg)
was remarkably decreased in the magnesium gluconate ion at m/z 447 in the treated sample by 78.26% compared with the
control sample. Consequently, the isotopic abundance ratio of P
M+1
/P
M
(
2
H/
1
H or
13
C/
12
C or
17
O/
16
O) in gluconate ion in the
treated sample was significantly increased by 37.35% with respect to the control sample. Thus, the Trivedi Effect
®
treated
magnesium gluconate could be valuable for designing better pharmaceutical and/or nutraceutical formulations through its
changed physicochemical and thermal properties, which might be providing better therapeutic response against various
diseases such as diabetes mellitus, allergies, aging, inflammatory diseases, immunological disorders, and other chronic
infections. The treated magnesium gluconate might be helpful to design the novel potent enzyme inhibitors by using its kinetic
isotope effects.
125 Mahendra Kumar Trivedi et al.: Structural and Natural Isotopic Abundance Analysis of Magnesium Gluconate
Treated with Energy of Consciousness (The Trivedi Effect®) Using LC-MS and NMR
Keywords: Consciousness Energy Healing Treatment, Biofield Energy Healers, The Trivedi Effect
®
,
Magnesium Gluconate, LC-MS, NMR, Isotopic Abundance Ratio, Isotope Effects
1. Introduction
The Magnesium ion (Mg
2+
) is a major intracellular
divalent cation. It is an essential mineral for several enzymes,
DNA and RNA synthesis, reproduction and protein synthesis
as well as a vital coherent controller of glycolysis and the
Krebs cycle [1, 2]. Magnesium gluconate (C
12
H
22
MgO
14
) is
the organometallic salt of magnesium with gluconic acid
produced from glucose catalyzed by glucose oxidase [3].
Magnesium gluconate is found to be the most powerful
antioxidant than other magnesium salts and it is useful for the
prevention and treatment of many diseases such as
cardiovascular diseases, diabetes, allergies, inflammatory
diseases, immunological disorders, Alzheimers disease,
asthma, pre-eclampsia and eclampsia, cancer, etc. [4-8]. It
can be used as neuroprotective [9], for the treatment of
oxidative stress induced ischemia/reperfusion injury [10] and
also labor in women arrested initially with intravenous
therapy as an oral tocolytic agent [11]. Magnesium gluconate
showed the highest bioavailability and most physiologically
acceptable salt among other magnesium salts like chloride,
sulfate, carbonate, acetate, citrate, lactate, aspartate, etc. [8,
12]. This treated magnesium gluconate mineral can be for the
prevention and treatment of various human diseases.
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 correlate 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, paradimensional and
can freely flow between the human and environment [13,
14]. So, a human has the ability to harness energy from the
ionosphere of the earth, the “universal energy field”, and
transmit it to any living organism(s) or nonliving object(s)
around the globe. The object or recipient always receives the
energy and responds in a useful way. This process is known
as The Trivedi Effect
®
- Biofield Energy Healing Treatment
[15, 16]. 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) [17]. Biofield Energy Treatment (The Trivedi
Effect
®
) has been extensively studied with significant
outcomes in many scientific fields such as cancer research
[18, 19], altered antimicrobial sensitivity of pathogenic
microbes in microbiology [20, 21], genetics [22, 23],
biotechnology [24, 25], altered physical and chemical
properties of pharmaceuticals [26, 27], nutraceuticals [28,
29], improved overall growth and yield of plants in
agricultural science [30, 31], and changing the structure of
the atom in relation to various metals, ceramics, polymers
and chemicals in materials science [32-35]. 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, 37]. The stable isotope ratio analysis has the
wide applications in several scientific fields for
understanding the isotope effects resulting from the variation
of the isotopic composition of the molecule [38, 39].
Conventional mass spectrometry (MS) techniques such as
liquid chromatography – mass spectrometry (LC-MS), gas
chromatography mass spectrometry (GC-MS) are widely
used for isotope ratio analysis with sufficient precision [40].
Hence, LC-MS and NMR (Nuclear Magnetic Resonance)
were used in this study to characterize the structural
properties of the Biofield Energy Treated and untreated
magnesium gluconate for the quality control purpose in
pharmaceutical and nutraceutical industries. Consequently,
LC-MS based isotopic abundance ratio (P
M+1
/P
M
and
P
M+2
/P
M
) analysis in both The Trivedi Effect
®
- treated and
untreated samples was aimed to investigate the influence of
The Trivedi Effect
®
- Consciousness Energy Healing
Treatment on the isotopic abundance ratio in magnesium
gluconate.
2. Materials and Methods
2.1. Chemicals and Reagents
Magnesium gluconate hydrate was purchased from Tokyo
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
Chemical and Biomolecular Engineering 2017; 2(3): 124-134 126
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 was
treated with the Energy of Consciousness by eighteen
renowned Biofield Energy Healers (The Trivedi Effect
®
) and
defined as Biofield Energy Treated or The Trivedi Effect
®
treated sample, while the second part of the test compound
did not receive any sort of treatment and was defined as the
untreated or control magnesium gluconate sample. The
Trivedi Effect
®
Treatment was provided by the group of
eighteen renowned The Trivedi Effect
®
Energy Healers who
participated in this study and performed the Trivedi Effect
®
Energy Treatment remotely. Eleven Energy Healers were
remotely located in the U.S.A., four remotely located in
Canada, two remotely located in Finland, and one of which
was remotely located in Albania, while the test compound
was located in the research laboratory of GVK Biosciences
Pvt. Ltd., Hyderabad, India. This Trivedi Effect
®
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
Energy Healers in this study visited the laboratory in person,
nor had any contact with the compounds. Similarly, the
control compound was subjected to a “sham” healer for 5
minutes, under the same laboratory conditions. The sham
healer did not have any knowledge about The Trivedi
Effect
®
- Energy of Consciousness Healing Treatment. After
that, the Trivedi Effect
®
Energy 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 Trivedi Effect
®
Energy
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 and negative 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 the scientific literature reported [41, 42] 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
Mg
26
Mg 26 11.01 13.94 n
Mg
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
achieved from the scientific literature and presented in the
Table 1 [43, 44].
127 Mahendra Kumar Trivedi et al.: Structural and Natural Isotopic Abundance Analysis of Magnesium Gluconate
Treated with Energy of Consciousness (The Trivedi Effect®) Using LC-MS and NMR
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] (1)
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 the control and Trivedi
Effect
®
Biofield Energy Treated magnesium gluconate are
presented in the Figure 1.
Figure 1. Liquid chromatograms of the control and Trivedi Effect
®
Biofield Energy Treated magnesium gluconate.
The ESI-MS spectra of the control and treated magnesium
gluconate at corresponding the retention time (R
t
) are shown
in the Figures 2 and 3. The R
t
of the mixture of water and
methanol (60:40 v/v) was at 2.35 min. Beside this, the liquid
chromatogram of the control sample showed two peaks
indicating that at R
t
of 1.81 and 2.06 min with the peak
area% of 61.22% and 38.78%, respectively (Figure 1). On
the other hand, the treated sample exhibited also two peaks at
R
t
of 1.79 and 2.03 min with peak area% of 60.65% and
39.35%, respectively (Figure 1). These findings indicated
that the polarity/affinity of the treated sample was unaltered
as compared to the control sample. The ESI-MS spectra of
the control and treated magnesium gluconate at R
t
of 2.06
and 2.03 min (Figure 2) indicated the presence of the mass of
magnesium gluconate adduct with methanol at m/z 447 [M +
CH
3
OH + H]
+
(calcd for C
13
H
27
MgO
15
+
, 447).
Figure 2. The ESI-MS spectra (Positive ionization mode) of the control and Biofield Energy Treated magnesium gluconate at the retention time 2.06 and 2.03
min, respectively.
Chemical and Biomolecular Engineering 2017; 2(3): 124-134 128
Figure 3. The ESI-MS spectra (Negative ionization) of the control and Biofield Energy Treated magnesium gluconate at the retention time 1.81 and 1.79 min,
respectively.
There was also a peak for the protonated molecular ion at m/z 415 (calcd for C
12
H
23
MgO
14
+
, 415) for magnesium gluconate
(Figure 2).
Figure 4. Proposed fragmentation pathway of magnesium gluconate.
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