Comprehensive Analytical Study of Consciousness Energy Treatment on Tocopherol Using GC-MS LC-MS FT-IR UV-Vis and NMR Spectroscopy

four tocopherols and four tocotrienols. The α - (alpha), β - (beta), γ - (gamma) and δ -(delta) forms of tocopherols and tocotrienols occur depend upon the number and position of methyl groups on the chromanol ring [1]. Alpha ( α )-tocopherol, the fully methylated tocopherol 1), is the most biologically active and abundant of all the components Abstract Alpha-tocopherol is the biologically active and abundant form of vitamin E found in mammalian tissues. Some of the major issues associated with tocopherol is solubility in solvents, sensitivity to oxidizing agents and air, critical storage conditions, and the absorption and efficiency in the body are widely variable. Therefore, the current study was designed to investigate the impact of the Trivedi Effect ® -Consciousness Energy Treatment on the isotopic abundance ratios (P M+1 /P M and P M+2 /P M ) and structural properties of tocopherol using LC-MS, GC-MS, FT-IR, UV-vis, and NMR spectroscopy. The Tocopherol sample was divided into two parts. One part of the sample was termed as the untreated/control sample, while the other part of the sample

of vitamin E found in mammalian tissues [2]. The biological function of α-tocopherol is more interesting on membrane-resident proteins and enzymes, gene expression, and signaling cascades.
It may be related to its contribution to membrane structure and dynamics, and thus modulation of membrane dependent signaling mechanisms such as protein and lipid kinases [3]. Vitamin E is used mainly as an antioxidant in preparations containing fat (ointments, creams, parentrals, oils, etc.) and cosmetics [4]. In combination with other antioxidants, such as zinc and vitamin C, vitamin E indicates a protective against the onset and progression of agerelated macular degeneration (AMD) [5]. It is also involved in the regulation of the production of eicosanoids by inhibition of both phospholipases A 2 (PLA 2 ) and cyclooxygenase (COX) activities. It has been used to slow down the progression of cataracts, coronary heart disease, oxidative stress leading to pre-eclampsia, Alzheimer's and Parkinson's, and other degenerative diseases [6][7][8][9][10]. The combined α and γ tocopherol supplements may be considered in prostate and other cancer prevention [11]. Deficiency of tocopherol can cause spinocerebellar ataxia [12], myopathies [13], peripheral neuropathy, ataxia, skeletal myopathy, retinopathy, impairment of the immune response [14], red blood cell destruction [15], etc.
Chronic use of high doses of tocopherol may cause nausea, diarrhea, or vision deficiencies [16]. Some of the major issues associated with tocopherol is the solubility in solvents, high sensitivity to oxidizing agents, turning dark on exposure to air, critical storage conditions, and the absorption and efficiency in the body are widely variable [17,18]. The Trivedi Effect ® -Energy of Consciousness Treatment has significantly influenced the bioavailability of poorly bioavailable compounds (i.e., resveratrol, berberine, and 25-hydroxyvitamin D3 in Male Sprague-Dawley rats) [15][16][17] and altered the physicochemical and thermal properties of many pharmaceutical/ nutraceutical compounds [19][20][21][22][23][24][25]. The Trivedi Effect ® is natural and the only scientifically proven phenomenon in which a person can harness this inherently intelligent energy from the universe and transmit it anywhere on the planet [26]. Biofield Energy is the electromagnetic field exist around the human body [27,28]. Biofield based Energy Therapies are also used against various human disease conditions and accepted worldwide [27][28][29] [34,35], crops [36,37], microbes [38,39], etc. The Trivedi Effect ® , assumed to act through the possible mediation of neutrinos [26], has also altered the isotopic abundance ratio of some of the organic compounds [34,35]. A study on the natural stable isotope is required to understand the isotope kinetic effects resulting from the alterations of the isotopic composition, which have many applications in different fields of sciences [40][41][42]. Highly sophisticated analytical techniques such as Gas chromatography -mass spectrometry (GC-MS) and liquid chromatography -mass spectrometry (LC-MS) are widely used for the study of isotopic abundance ratio analysis with sufficient precision [41]. Therefore, a study has been performed to determine the impact of the Trivedi Effect ® -Consciousness Energy Treatment on the isotopic abundance ratios of P M+1 /P M ( 2 H/ 1 H or 13

Chemicals and reagents
D/L-alpha-tocopherol (99.30%), a clear yellow/brown viscous oil was purchased from Alfa Aesar, India, and the other chemicals used in the experiment were of analytical grade also available in India.

Consciousness energy healing treatment strategies
The tocopherol test sample was equally divided into two parts.
The mass spectrometric analysis was performed under the +ve ESI mode. The total ion chromatogram peak area% and mass spectrum of the individual peak which appeared in LC, along with the full scan, were recorded. The mass peak intensities of the mass spectrum of the individual peak were recorded. The natural abundance of the C, O, and H isotopes can be predicted from the comparison of the relative abundance of the isotope peak with respect to the base peak. The values of the natural isotopic abundance of the common elements were obtained from the literature [42][43][44][45]. The isotopic abundance ratios (P M+1 /P M and P M+2 /P M ) for the Control and the Biofield Energy Treated tocopherol were calculated. % change in isotopic abundance ratio of the Treated tocopherol = [(IAR Treated -IAR Control )/ IAR Control ) x 100] Where, IAR Treated : isotopic abundance ratio in the Treated tocopherol and IAR Control : isotopic abundance ratio in the Control tocopherol.

Gas chromatography-mass spectrometry (GC-MS) analysis
The GC-MS analysis of the Control and the Biofield Energy Treated tocopherol was performed using Agilent 7890B with 5977B Mass selective detector, USA [46]. Gas chromatograph equipped with a silica capillary column HP-5 MS (30m x 0.25mm x 0.25µm) and coupled to the quadrupole detector with pre-filter.

Fourier transform infrared (FT-IR) spectroscopy
The FT-IR spectroscopy was recorded on the Spectrum ES (Perkin Elmer, USA) Fourier transform infrared spectrometer with the frequency array of 400-4000cm -1 . The compound was run as NaCl disks in mull method (2mg of Control/Biofield Energy Treated sample in 2 drops of chloroform).

Ultraviolet-visible pectroscopy (UV-Vis) analysis
The UV-Vis spectral analysis of the tocopherol was carried out using a Shimadzu UV-2400PC series (Japan). The absorbance spectra with the wavelength of maximum absorbance (λ max) were recorded.

Isotopic abundance ratio analysis
The alteration in isotopic abundance ratio may affect the kinetic isotope effects of the atoms/molecules. It is very useful to study the reaction mechanism, understand the enzymatic transition state, and the mechanism that is supportive for designing effective and specific inhibitors, etc. [42]. Therefore, the Biofield Energy Treated tocopherol with improved isotopic abundance ratio (P M+1 /P M and P M+2 /P M ) was assumed to be more advantageous for the designing of better nutraceutical/pharmaceutical formulations.

Gas Chromatography-mass spectrometry (GC-MS) analysis
The  Table 2). The increased peak area of the Biofield Energy Treated tocopherol is assumed to be the influence of Consciousness Energy Treatment (the Trivedi Effect ® ).

Fourier transform infrared (FT-IR) spectroscopy
The Control and the Biofield Energy Treated tocopherol samples were investigated by FT-IR spectroscopy ( Figure 6). The

FT-IR spectra of both the Control and the Biofield Energy Treated
Tocopherol showed the clear stretching and bending peak in the functional group and fingerprint region. The broad peak in the functional group region at 3472cm -1 in the Control and 3478cm -1 in the Biofield Energy Treated spectrum was due to O-H stretching.
The spectra showed aliphatic C-H stretching at 2929, and 2868cm -1 for both the Control and the Biofield Energy Treated samples

Ultraviolet-visible spectroscopy (UV-Vis) analysis
The UV-visible spectra of the Control and the Biofield Energy

Nuclear magnetic resonance (NMR) spectroscopy analysis
The 1 H NMR spectra of the Control and the Biofield Energy Treated tocopherol are shown in Figure 8. The signals for the proton coupling of CH 3 , CH 2 , CH, and OH protons in both the 1 H NMR spectra of tocopherol were in the range of δ 0.8 to 7.4ppm ( Figure 8 and Table   3). The 1 H signals for the Control and the Biofield Energy Treated tocopherol were close to each other. Correspondingly, 13 C NMR spectra of the Control and the Biofield Energy Treated tocopherol are shown in Figure 9. The carbon signals for CH 3 , CH 2 , CH, =C<, and C-OH groups in both the Control and the Biofield Energy Treated NMR spectra were in the range of 11.5-145.1 (Figure 9 and Table   3). The 13 C signals for the Control and the Biofield Energy Treated tocopherol were close to each other. The experimental data were closely matched to the published literature. The NMR spectral data indicated that structure of the Biofield Energy Treated tocopherol did not alter compared to the Control sample.

Conclusion
Overall experimental results revealed that the Trivedi Effect ® -

Consciousness Energy Treatment (Biofield Energy Treatment)
showed a significant impact on the mass peak intensities, isotopic abundance ratios and the peak area of tocopherol. The LC-ESI-MS analysis of both the samples showed the mass of protonated parent molecular ion at m/z 431.3 (calcd for C 29