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Open Access Journal of Cancer & Oncology Research Article 16 min read

Isotopic Abundance Ratio Analysis of the Consciousness Energy Healing Treated Flutamide using LC-MS and GC-MS Spectrometry

Snehasis Jana
ISSN: 2578-4625  10.23880/oajco-16000167  Received: January 21, 2021  Published: February 16, 2021
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Keywords
Flutamide The Trivedi Effect® Biofield Energy Consciousness Energy LC-MS GC-MS
Abstract

Flutamide is used to treat prostate cancer, which acts by blocking the action of exogenous testosterone binding through its androgen receptor. This study was designed and performed to investigate the impact of the Trivedi Effect®-Consciousness Energy Healing Treatment on the structural properties and the isotopic abundance ratio of flutamide using LC-MS and GC-MS spectroscopy. Flutamide sample was divided into control and treated parts. Only the treated flutamide received the Trivedi Effect®-Consciousness Energy Healing Treatment remotely by a famous Biofield Energy Healer, Dahryn Trivedi. The LC-MS spectra of both the samples at retention time (Rt ) 3.4 minutes exhibited the mass of the deprotonated molecular ion peak at m/z 275.08 [M-H]-. The peak area of the treated sample was significantly increased by 92.42% compared to the control sample, which indicated that the solubility profile of the treated sample was significantly increased compared to the control sample. The LC-MS based isotopic abundance ratio of PM+1/PM (2 H/1 H or 13C/12C or 15N/14N or 17O/16O) in the treated flutamide was significantly increased by 59.48% compared with the control sample. The GC-MS based isotopic abundance ratio of PM+1/ PM in the treated flutamide sample was increased by 10.39% compared with the control sample. The results indicated that the 13C, 2 H, 15N, and 17O contributions from (C11H11F3 N2 O3 )+ to m/z 277 in the treated sample were significantly increased compared with the control sample. The isotopic abundance ratio of PM+1/PM (2 H/1 H or 13C/12C or 15N/14N or 17O/16O) in the treated flutamide was significantly increased compared to the control sample. The changes in the peak area and isotopic abundance could be due to changes in nuclei possibly through the interference of neutrino via the Trivedi Effect®-Consciousness Energy Healing Treatment. The new form of treated flutamide was formed that might have increase the chemical bond strength, stability, solubility, bioavailability, and efficacy, which could be very much useful to design more efficacious pharmaceutical formulations against prostate cancer, androgen-dependent skin and hair conditions including acne, seborrhea, hirsutism, and scalp hair loss, hyperandrogenism, as well useful for feminizing hormone therapy aimed at transgender women.

Introduction

Flutamide is a nonsteroidal antiandrogen (NSAA) which blocks the action of both endogenous and exogenous testosterone by inhibiting the androgen receptor. Further, it is a potent inhibitor of testosterone-stimulated prostatic DNA synthesis and it is capable of inhibiting prostatic nuclear uptake of androgen [1, 2, 3]. Flutamide can be used independently for the treatment or in combination with other medications and radiation treatments [4]. It is used primarily to treat men with prostate cancer [5]. The testosterone hormone aids prostate cancer to grow and spread [6]. It is also used in the treatment of androgen-dependent problems, i.e., acne, seborrhea, hirsutism, scalp hair loss, and hyperandrogenism. It can be used as a constituent of feminizing hormone therapy for transgender women [7]. Overdose of flutamide may cause anorexia, ataxia, piloerection, hypoactivity, slow respiration, and/or lacrimation, emesis, tranquilization, and methemoglobinemia [8].

The physicochemical properties of a pharmaceutical compound in the formulation is very much important and decide the rate of dissolution, absorption, bioavailability, and efficacy in the body [9]. It was observed that the Trivedi Effect®-Consciousness Energy Healing Treatment (Biofield Energy Healing Treatment) has the significant impact on solubility and bioavailability by directly altering the isotope composition, physicochemical and thermal behaviours of various nutraceutical and pharmaceutical compounds [10, 11, 12]. The Trivedi Effect® is a natural and only technically established phenomenon in which an individual can harness this inherently intelligent energy from the “Universal Energy Field” and transfer it anywhere on the planet via the possible mediation of neutrinos [13]. “Biofield” is the electromagnetic energy field which exists surrounding the living organisms, generated by the continuous movement of the electrically charged particles (ions, cells, etc.) inside the body. It can transmit the electromagnetic energy in the form of bio-photons and this process is called Biofield Energy Healing Treatment [14, 15]. Biofield Therapies (Energy Medicine) have been reported with significant outcomes against various disease conditions [16]. The National Center for Complementary and Alternative Medicine (NCCAM) has recognized and accepted Biofield Energy Healing as a Complementary and Alternative Medicine (CAM) health care approach along with other therapies, medicines, and practices such as yoga, Qi Gong, Tai Chi, hypnotherapy, Reiki, etc. [17]. These CAM therapies have been accepted by most of the U.S.A. population with several advantages [18].

The significant outcome of the Trivedi Effect®- Consciousness Energy Healing Treatment has been widely reported. It has been reported altering the characteristic properties of the several non-living materials and living object(s), i.e., organic compounds [19, 20], metals and ceramic [21, 22], polymer [23], crops [24], nutraceuticals [25, 26], microbes [27, 28], etc.

The stable isotope ratio and its analysis have various applications in different field of science for understanding the isotope effects resulting from the variation of the isotopic composition of the molecule [29, 30]. Isotope ratio analysis can be performed by using the conventional mass spectrometry (MS) techniques such as gas chromatography- mass spectrometry (GC-MS) and liquid chromatography- mass spectrometry (LC-MS) in low micromolar concentration with sufficient precision [30, 31]. The Trivedi Effect®- Consciousness Energy Healing Treatment could be an economical approach for designing better pharmaceuticals formulations. Therefore, in this study, special attention was taken to improve the physicochemical parameters of the pharmaceutical product, e.g., flutamide. Hence, LC-MS and GC-MS were used in this study to characterize the structural properties and evaluate the isotopic abundance ratio analysis of PM+1/PM (2H/1H or 13C/12C or 15N/14N or 17O/16O) in the Trivedi Effect®-Consciousness Energy Healing Treated flutamide compared to the control sample.

Materials and Methods

Chemicals and Reagents

The test sample flutamide was purchased from Tokyo Chemical Industry Co., Ltd., Japan. Whereas, other chemicals and solvents used during the experiments were of analytical grade purchased in India.

Consciousness Energy Healing Treatment Strategies

The flutamide powder sample was divided into two parts. One part of flutamide powder sample was considered as a control sample, which did not receive the Biofield Energy Treatment. Further, the control flutamide was treated with a “sham” healer, did not have any knowledge about the Biofield Energy and its treatment procedure. However, the other part of flutamide was received the Trivedi Effect®-Consciousness Energy Healing Treatment remotely under standard laboratory conditions for 3 minutes by the renowned Biofield Energy Healer, Dahryn Trivedi, USA, and known as the Biofield Energy Treated flutamide. After completion of the treatment by the Biofield Energy Healer and “sham” healer, both the samples were kept in sealed conditions and characterized using sophisticated analytical techniques.

Characterization

Liquid Chromatography-Mass Spectrometry (LC-MS) analysis and calculation of iIsotopic abundance ratio: The LC-MS analysis of the control and Biofield Energy Treated flutamide was carried out with the help of LC-MS Thermo Fisher Scientific, the USA equipped with an ion trap detector connected with a triple-stage quadrupole mass spectrometer. The column used here was a reversed phase Thermo Scientific Synchronis C18 (Length-250 mm X ID 4.6 mm X 5 micron), maintained at 25˚C. Methanol was the diluent for the sample preparation. 10 µL of flutamide solution was injected, and the analyte was eluted using 92% acetonitrile + 8% 10 mM ammonium acetate pumped at a constant flow rate of 1 mL/min. Chromatographic separation was achieved using gradient condition and the total run time was 10 min. Peaks were monitored at 300 nm using the PDA detector and were performed under -ve ESI mode. The total ion chromatogram, peak area% and mass spectrum of the individual peak which was appeared in LC along with the full scan (m/z 50-500) were recorded.

The natural abundance of each isotope (C, O, H, N, and F) can be predicted from the comparison of the height of the isotope peak with respect to the base peak. The values of the natural isotopic abundance of the common elements are obtained from the literature [30, 32, 33, 34]. The LC-MS based isotopic abundance ratios (PM+1/PM) for the control and Biofield Energy Treated flutamide was calculated using equation 1.

% Change in isotopic abundance ratio = [(IARTreated – IARControl)/ IARControl] x 100 (1) Where IARTreated = isotopic abundance ratio in the treated flutamide and IARControl = isotopic abundance ratio in the control flutamide.

Gas Chromatography-Mass Spectrometry (GC-MS) analysis: GC-MS of the control and treated flutamide were analyzed with the help of Perkin Elmer Gas chromatograph equipped with a PE-5MS (30M x 250 micros x 0.250 microns) capillary column and coupled to a single quadrupole mass detector was operated with electron impact (EI) ionization (+ve ion mode). The oven temperature was programmed from 80°C (14 min hold) to 250°C (3 min hold) @ 10°C / min (total run time 25 min). The sample was prepared taking 60 mg of the flutamide is in 2 mL methanol as a diluent. Mass spectra were scanned from m/z 20 to 400. The identification of analyte was done by GC retention times and by a comparison of the mass spectra of samples. The GC-MS based isotopic abundance ratios (PM+1/PM) for the control and treated flutamide was calculated.

Results and Discussion

Liquid Chromatography-Mass Spectrometry (LC-MS)

The chromatograms and mass spectra of both the samples of flutamide are shown in Figures 1 and 2, respectively. The control and Biofield Energy Treated flutamide showed the single major chromatographic peak at the retention time (Rt) of 3.39 and 3.4 minutes in the chromatograms (Figure 1). The peak area of the Biofield Energy Treated flutamide was significantly increased by 92.42% compared to the control sample, which indicated that the solubility profile of the Biofield Energy Treated sample was significantly increased after the Biofield Energy Treatment compared to the control sample.

Figure 1: Liquid chromatograms of the control and Biofield Energy Treated flutamide.
Click to enlarge
Figure 1: Liquid chromatograms of the control and Biofield Energy Treated flutamide.

The molecular mass peak [M]+ of flutamide found to be at m/z 276 in the mass spectrum [35]. In this case, the mass spectra of both the samples (Figure 2) showed the deprotonated molecular ion peak at m/z 275.08 [M-H]- (calculated for C11H10F3N2O3 -, 175.06) along with other fragmentation peaks (Figure 2).

Figure 2: Mass spectra of the control and Biofield Energy Treated flutamide at Rt 3.4 minutes.
Click to enlarge
Figure 2: Mass spectra of the control and Biofield Energy Treated flutamide at Rt 3.4 minutes.

The LC-MS spectra of both the samples showed the mass of the molecular ion peak at m/z 275.08 [M-H]- (calculated for C11H10F3N2O3 -, 275.06) with relative intensity of 100%.

The theoretical calculation of PM+1 for flutamide was presented as below:

P (13C) = [(11 x 1.1%) x 100% (the actual size of the M- peak)] / 100% = 12.1%

P (2H) = [(10 x 0.015%) x 100%] / 100%= 0.15% P (15N) = [(2 x 0.4%) x 100%] / 100% = 0.8% P (17O) = [(3 x 0.04%) x 100%] / 100% = 0.12% PM+1, i.e. 13C, 2H, 15N, and 17O contributions from (C11H10F3N2O3)- to m/z 276.08 = 13.17% From the above calculation, it has been found that 13C and 15N have major contributions from (C11H10F3N2O3)- to m/z 276.08.

Control sampleBiofield Energy Treated sample
$P_{M at m/z 275.08 (%)100100
PM+1 at m/z 276.08 (%)8.3913.41
PM+1/PM}$$0.080.13
% Change of isotopic abundance ratio (PM+1/PM) with respect to the control sample59.48

Table 1: LC-MS based isotopic abundance analysis results in Biofield Energy Treated flutamide compared to the control sample. PM:

The LC-MS based isotopic abundance ratio analysis PM and PM+1 for flutamide near m/z 275.08 and 276.08, respectively of the control and Biofield Energy Treated samples, which were obtained from the observed relative peak intensities of [M-] and [(M+1)-] peaks, respectively (Table 1). The % change of the isotopic abundance ratio (PM+1/PM) in the treated flutamide was significantly increased by 59.48% compared with the control sample (Table 1). Therefore, it was concluded that the 13C, 2H, 15N, and 17O contributions from (C11H10F3N2O3)- to m/z 276.08 in the treated flutamide were significantly increased compared to the control sample.

Gas Chromatography-Mass Spectrometry (GC- MS) Analysis

The GC-MS chromatograms showed the presence of a single chromatographic peak at the retention time of 18.8 minutes in both the samples of flutamide (Figures 3 and 4). The parent molecular ion peak of flutamide at m/z 276 [M]+ (calculated for C11H11F3N2O3 +, 276.07) in the control sample and Biofield Energy Treated sample, along with the fragment ion peaks near m/z 246, 233, 206, 187, 71, and 43 (Figures 3 and 4) which corresponded to the molecular formula C10H9F3N2O2 2+, C9H8F3N2O2 +, C7H5F3N2O2 2+, C7H5F2N2O2 +, C4H7O+, and C3H7 +, respectively were proposed (Figure 5).

The GC-MS spectra of both the control and Biofield Energy Treated flutamide showed the mass of the molecular ion peak [M]+ at m/z 276 (calculated for C11H11F3N2O3 +, 276.07).

The theoretical calculation of PM+1 for flutamide was presented as below:

P (13C) = [(11 x 1.1%) x 2.62% (the actual size of the M+ peak)] / 100% = 0.32%

P (2H) = [(11 x 0.015%) x 2.62%] / 100%= 0.004%

P (15N) = [(2 x 0.4%) x 2.62%] / 100% = 0.02%

P (17O) = [(3 x 0.04%) x 2.62%] / 100% = 0.003%

PM+1, i.e. 13C, 2H, 15N, and 17O contributions from (C11H11F3N2O3)+ to m/z 277 = 0.37% The calculated isotope abundance (0.37%) was close to the experimental value 0.32% (Table 2). From the above calculation, it has been found that 13C and 15N have major contribution to m/z 277.

Figure 3: The GC-MS chromatogram and mass spectra of the control flutamide.
Click to enlarge
Figure 3: The GC-MS chromatogram and mass spectra of the control flutamide.
Figure 4: The GC-MS chromatogram and mass spectra of the Biofield Energy Treated flutamide.
Click to enlarge
Figure 4: The GC-MS chromatogram and mass spectra of the Biofield Energy Treated flutamide.
Figure 5: Proposed fragmentation pattern of flutamide.
Click to enlarge
Figure 5: Proposed fragmentation pattern of flutamide.
ParameterControl sampleBiofield Energy Treated sample
P at m/z 276 (%)
M		2.621.78
P at m/z 277 (%)
M+1		0.320.24
P /P
M+1 M		0.120.13
% Change of isotopic abundance ratio (P /P ) with
M+1 M
respect to the control sample		10.39

The GC-MS based isotopic abundance ratio of PM+1/PM in the Biofield Energy Treated flutamide was significantly increased by 10.39% compared with the control sample (Table 2). Hence, 13C, 2H, 15N, and 17O contributions from (C11H11F3N2O3)+ to m/z 277 in the Biofield Energy Treated sample were increased compared with the control sample.

The data confirmed the structure of the sample as flutamide. The peak area and isotopic abundance ratio of PM+1/PM (2H/1H or 13C/12C or 15N/14N or 17O/16O) in the Biofield Energy Treated flutamide were significantly increased compared to the control sample. According to modern physics, the neutrino is an elementary particle which changes identities. It is only possible if the neutrinos possess mass and have the ability to interchange their phase from one phase to another internally. Therefore, the neutrinos have the capability to interact with protons and neutrons in the nucleus, which indicated a close relation between neutrino and the isotope formation [12, 30, 31]. The altered isotopic composition in the molecular level of the treated flutamide might have altered the neutron to proton ratio in the nucleus. It can be assumed that the alterations in the isotopic abundance could be due to the change in nuclei by the interference of neutrino particles via the Trivedi Effect®- Consciousness Energy Healing Treatment.

The improved isotopic abundance ratios 2H/1H or 13C/12C or 15N/14N or 17O/16O would highly influence the atomic bond vibration of treated flutamide. The increased isotopic abundance ratio of the Consciousness Energy Healing Treated flutamide would stronger the chemical bond and increase the stability [36, 37]. The Trivedi Effect®- Consciousness Energy Healing Treated flutamide might improve the solubility, bioavailability, and therapeutic efficacy compared to the control sample. The new form of Biofield Energy Treated flutamide would be very much useful to design better pharmaceutical formulations against prostate cancer, androgen-dependent skin and hair conditions including acne, seborrhea, hirsutism, and scalp hair loss, hyperandrogenism, as well useful for feminizing hormone therapy aimed at transgender women.

Conclusion

The Trivedi Effect®-Consciousness Energy Healing Treatment showed a significant impact on the isotopic abundance ratios and peak area of flutamide. The LC-MS spectra of both the samples at retention time (Rt) 3.4 minutes exhibited the mass of the deprotonated molecular ion peak at m/z 275.08 [M-H]-. The peak area of the Biofield Energy Treated flutamide was significantly increased by 92.42% compared to the control sample, which indicated that the solubility profile of the Biofield Energy Treated flutamide was significantly increased compared to the control sample.

The LC-MS based isotopic abundance ratio of PM+1/PM (2H/1H or 13C/12C or 15N/14N or 17O/16O) in the Biofield Energy Treated flutamide was significantly increased by 59.48% compared with the control sample. The GC-MS based isotopic abundance ratio of PM+1/PM in the Biofield Energy Treated flutamide was increased by 10.39% compared with the control sample. The results indicated that the 13C, 2H, 15N, and 17O contributions from (C11H11F3N2O3)+ to m/z 277 in the Biofield Energy Treated flutamide were significantly increased compared with the control sample. The isotopic abundance ratio of PM+1/PM (2H/1H or 13C/12C or 15N/14N or 17O/16O) in the Biofield Energy Treated flutamide was significantly increased compared to the control sample. The changes in the peak area and isotopic abundance could be due to changes in nuclei possibly through the interference of neutrino via the Trivedi Effect®-Consciousness Energy Healing Treatment. The new form of treated flutamide was formed that might have increase the chemical bond strength, stability, solubility, bioavailability, and efficacy, which could be very much useful to design more efficacious pharmaceutical formulations against prostate cancer, androgen-dependent skin and hair conditions including acne, seborrhea, hirsutism, and scalp hair loss, hyperandrogenism, as well useful for feminizing hormone therapy aimed at transgender women.

Acknowledgement

The authors are grateful to Sophisticated Instrumentation Centre for Applied Research & Testing (SICART) India, Trivedi Science, Trivedi Global, Inc., Trivedi Testimonials, and Trivedi Master Wellness for their assistance and support during this work.

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@article{snehasis2021,
  title   = {Isotopic Abundance Ratio Analysis of the Consciousness Energy
Healing Treated Flutamide using LC-MS and GC-MS Spectrometry},
  author  = {Snehasis Jana},
  journal = {Open Access Journal of Cancer & Oncology},
  year    = {2021},
  volume  = {5},
  number  = {1},
  doi     = {10.23880/oajco-16000167}
}
Snehasis Jana (2021). Isotopic Abundance Ratio Analysis of the Consciousness Energy
Healing Treated Flutamide using LC-MS and GC-MS Spectrometry. Open Access Journal of Cancer & Oncology, 5(1). https://doi.org/10.23880/oajco-16000167
TY  - JOUR
TI  - Isotopic Abundance Ratio Analysis of the Consciousness Energy
Healing Treated Flutamide using LC-MS and GC-MS Spectrometry
AU  - Snehasis Jana
JO  - Open Access Journal of Cancer & Oncology
PY  - 2021
VL  - 5
IS  - 1
DO  - 10.23880/oajco-16000167
ER  -