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Abstract

Ascorbic acid is a water-soluble vitamin (Vitamin C) essential for both the plants and animals for the metabolic process. In this study, the liquid chromatography-mass spectrometry (LC-MS) analytical technique was used to characterize the structural properties and isotopic abundance ratio to evaluate the effect of the Trivedi Effect^®-Consciousness Energy Healing Treatment on L-ascorbic acid compared to the control sample. The ascorbic acid sample was divided into control and treated parts. Only the treated part received the Trivedi Effect^®-Consciousness Energy Healing Treatment remotely by a well-known Biofield Energy Healer, Mahendra Kumar Trivedi. The control and treated samples showed a chromatographic peak at retention time (R_t) 1.8 minutes exhibited the deprotonated molecular ion peak at m/z 175 (M-H)^- (calculated for C₆H₇O₆^-, 175.02) in the mass spectra. The peak area of the treated sample (12817614.01) was significantly increased by 8.81% compared to the control sample (11779918.9). The LC-MS based isotopic abundance ratio of P_M+1/P_M (²H/¹H or ¹³C/¹²C or ¹⁷O/¹⁶O) in the treated ascorbic acid was significantly increased by 23.22% compared with the control sample. Thus,13C, ²H, and17O contributions from (C₆H₇O₆)^- to m/z 176 in the treated ascorbic acid were significantly increased compared with the control sample. The increased isotopic composition of the treated ascorbic acid might have altered the neutron to proton ratio in the nucleus. The changes in isotopic abundance could be due to changes in nuclei possibly through the interference of neutrino particles via the Trivedi Effect^®-Consciousness Energy Healing Treatment. The increased isotopic abundance ratio and peak area of the treated ascorbic acid may increase the intra-atomic bond strength and its stability. This novel ascorbic acid after the Trivedi Effect^®-Consciousness Energy Healing Treatment would be very useful to design more efficacious pharmaceutical formulations against scurvy, obesity, cardiovascular diseases, hypertension, rheumatoid arthritis, Alzheimer's disease, cancer, etc.

Introduction

Ascorbic acid (Vitamin C) is required by both plants and animals for the essential metabolic process. Vitamin C is plenty available in citrus fruits and vegetables, red peppers, potatoes, milk, oysters, animal liver, etc. 12. Ascorbic acid converts to ascorbate at neutral pH or above pH 5 inside the cells and acts as a potent antioxidant 345. Vitamin C act as a cofactor in many enzymatic and non-enzymatic reactions. It helps in the synthesis of carnitine, collagen, and neurotransmitters, wound-healing and prevents bleeding from capillaries, metabolism of microsome, and synthesis and catabolism of tyrosine in the body 2. It is useful in the prevention and treatment of scurvy, hypertension, obesity, cancer, Alzheimer's disease, cardiovascular diseases, rheumatoid arthritis, etc. 3, 67891011. Deficiency of ascorbic acid leads to scurvy, gingivitis, autoimmune disease, nose bleeding, weak immune system, slow wound healing, dry and splitting hair, leaky gut, swollen and painful joints, cancer, high blood pressure, stroke, gallbladder disease, atherosclerosis, etc. 789. Low plasma concentrations of ascorbic acid are observed in the case of diabetes, infections, and smoking 12. Vitamin C supplements are available for the prevention and treatment of vitamin deficiency diseases. The tolerable upper intake level of ascorbic acid is 2,000 mg/day, and excess of ascorbic acid in the body may cause indigestion, diarrhoea, headache, fatigue, disturbed sleep, haemochromatosis, skin rashes, suppress the production of progesterone from the corpus luteum in healthy subjects 213.

The limitation associated with ascorbic acid is that it degrades while cooking, processing, packaging, storage, and cooking of blended foods 131415. Exposure to air and temperature significantly affects the stability of ascorbic acid 1415. Dissolution, absorption, bioavailability, and stability of ascorbic acid depend upon the physicochemical properties of any pharmaceutical compounds 17. Improvement of the physicochemical properties of pharmaceutical and nutraceutical products is a challenging task 18. But, it was observed that the Trivedi Effect^®-Biofield Energy Healing Treatment has a significant impact on the physicochemical properties and bioavailability of pharmaceutical / nutraceutical compounds 19202122. The Trivedi Effect^® is a natural and only scientifically established phenomenon in which an individual can harness this inherently intelligent energy and transmit it anywhere on the globe through the possible mediation of neutrinos 23. “Biofield Energy” the electromagnetic energy field which exists surrounding the living beings, which transmit electromagnetic energy in the form of bio-photons generated by the continuous movement of the electrically charged particles like ions, cells, blood flow, etc. inside the body. Biofield Energy Healing specialists have the ability to harness the energy from the Universe and can transmit into any living and non-living object( s). This process is called Biofield Energy Treatment 2425. Biofield based Energy Therapies have been reported with significant outcomes against various diseases, human health, and wellness 26. The National Center of Complementary and Integrative Health (NCCIH) has been recognized and accepted Biofield Energy Healing treatment as a Complementary and Alternative Medicine (CAM) health care approach along with other therapies, medicines, and practices 27. Most of the U.S.A. people have adopted these therapy 28. The Trivedi Effect^®-Consciousness Energy Healing Treatment has also reported with altering the physical and thermal properties of organic compounds, polymers, ceramics, and metals 293031323334, improved productivity of crops 35, and healing cancer 36.

The Trivedi Effect^®-Consciousness Energy Healing Treatment has a significant impact on the isotopic abundance of organic compounded 3334. The study of stable isotope ratio analysis has various applications in different fields of science 3738. The change in the isotopic abundance affects the physicochemical properties of the substance. The conventional liquid chromatography-mass spectrometry (LC-MS) can be useful for the study of isotopic abundance in low micromolar concentration with sufficient precision 3839. The Trivedi Effect^®-Biofield Energy Healing Treatment could be an economical approach for designing more efficacious pharmaceuticals formulations. Thus, the LC-MS was used to evaluate the impact of the Trivedi Effect^®-Consciousness Energy Healing Treatment on the structural properties and isotopic abundance ratio of L-ascorbic acid compared to the control sample.

Materials And Methods

Materials and Methods Chemicals and Reagents

The L-ascorbic acid (vitamin C) powder sample was purchased from Alfa Aesar, India. Similarly, other chemicals like acetonitrile and ammonium acetate were purchased from Merck India.

Consciousness Energy Healing Treatment Strategies

The vitamin C test sample powder was divided into control, and Biofield Energy Treated parts. The control sample did not receive the Biofield Energy Treatment but was treated with a sham healer who did not have any knowledge about the Biofield Energy Treatment. However, the treated ascorbic acid was received the Trivedi Effect^®-Consciousness Energy Healing Treatment remotely under standard laboratory conditions for 3 minutes by the renowned Biofield Energy Healer, Mahendra Kumar Trivedi, USA. After that, both the ascorbic acid samples were kept in sealed conditions and characterized using LC-MS analytical technique.

Characterization Liquid Chromatography-Mass Spectrometry (LC-MS) Analysis

The LC-MS analysis of ascorbic acid was carried out with the help of LC-MS ThermoFisher Scientific, the USA equipped with an ion trap detector connected with a triple-stage quadrupole mass spectrometer. A reversed-phase Thermo Scientific Synchronis C18 (Length-250 mm X ID 4.6 mm X 5 micron), maintained at 25˚C was used. The diluent used for the sample preparation was methanol and water. 10 µL of the ascorbic acid solution was injected, and the analyte was eluted using 80% acetonitrile + 20% 5 mM ammonium acetate pumped at a constant flow rate of 1 mL/min (total run time 10 min). Peaks were monitored at 220 nm using the PDA detector. The mass spectrometric analysis was performed under -ve ESI mode.

The values of the natural isotopic abundance of the common elements are obtained from the literature 384041. The LC-MS based isotopic abundance ratio (P_M+1/P_M) for both the ascorbic acid samples were calculated using equation (1).

% Change in isotopic abundance ratio = [(IAR_Treated - IAR_Control)/ IAR_Control)] x 100 (1)

Where IAR_Treated = isotopic abundance ratio in the treated sample and IAR_Control = isotopic abundance ratio in the control sample.

Results

Results and Discussion Liquid Chromatography-mass Spectrometry (LC-MS)

The chromatograms and mass spectra of both the control and Biofield Energy Treated samples of ascorbic acid are shown in Figure 1 and Figure 2, respectively. The chromatographic peak was observed at the retention time (R_t) of 1.8 minutes in the case of both the samples (Figure 1). But the peak area of the Biofield Energy Treated sample (12817614.01) was significantly increased by 8.81% compared to the control sample (11779918.9). One of the previous studies of the Biofield Energy Healer Mahendra Kumar Trivedi on ascorbic acid supported the results, in which the particle size of the treated ascorbic acid was reduced by 8.23% (d₁₀), 22.07% (d₅₀), 11.64% (d₉₀), and 15.81% {D(4,3)} and the surface area was significantly increased by 15.38% as compared to the untreated sample 42. This indicated that the solubility of the Biofield Energy Treated ascorbic acid was significantly increased.

Figure 1. Liquid chromatograms of the control and Biofield Energy Treated ascorbic acid. Figure 2. Mass spectra of the control and Biofield Energy Treated ascorbic acid at Rt 1.8 minutes.

The mass spectra of both the samples of ascorbic acid exhibited the mass of the deprotonated molecular ion ^M^- peak at m/z 175 (calculated for C₆H₇O₆^-, 175.02) along with other low molecular fragmentation peaks at 143, 115, 99, 87, and 70 for C₆H₇O₄^-, C₅H₇O₃^-, C₅H₇O₂^-, C₄H₇O₂^-, and C₄H₆O^•-(Figure 2 and Figure 3). The data also supported by the published literature 43.

Figure 3. Proposed fragmentation pattern of ascorbic acid in –ve ion mode.

The LC-MS spectra of both the ascorbic acid showed the mass of the molecular ion peak at m/z 175 ^M^- (calculated for C₆H₇O₆^-, 175.02) with 100% relative intensity. The theoretical calculation of P_M+1 for ascorbic acid was presented as below:

P(13C) = [(6 x 1.1%) x 100% (the actual size of the M⁺ peak)] / 100% = 6.6%

P(2H) = ^{(7 x 0.015%) x 100%} / 100%= 0.105%

P(17O) = ^{(6 x 0.04%) x 100%} / 100% = 0.24%

P_M+1,i.e.13C, 2H, and17O contributions from (C₆H₇O₆)^- to m/z 176 = 6.95%

From the above calculation, it has been found that 13C and 17O have a major contribution to m/z 176.

The LC-MS based isotopic abundance ratio analysis P_M (m/z 175) and P_M+1 (m/z 176) of both the samples were obtained from the observed relative peak intensities of [M⁺] and [(M+1)⁺] peaks, respectively in the mass spectra (Table 1). The percentage change of the isotopic abundance ratio (P_M+1/P_M) in the Biofield Energy Treated ascorbic acid was significantly increased by 23.22% compared to the control sample (Table 1). Therefore, the13C, 2H, and17O contributions from (C₆H₇O₆)^- to m/z 176 in the Biofield Energy Treated sample were significantly increased compared to the control sample.

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

Parameter	Control sample	Biofield Energy Treated sample
P_Mat m/z 175 (%)	100	100
P_M+1 at m/z 176 (%)	5.21	6.42
P_M+1/P_M	0.05	0.06
% Change of isotopic abundance ratio (P_M+1/P_M) with respect to the control sample		23.22

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)⁺], M: mass of the parent molecule.

The study confirmed the structure of the sample as ascorbic acid. The isotopic abundance ratio of P_M+1/P_M (2H/1H or 13C/12C or 17O/16O) in the Consciousness Energy Healing Treated ascorbic acid was significantly increased compared to the control sample. The increased isotopic composition of the Consciousness Energy Healing Treated ascorbic acid might have altered the neutron to proton ratio in the nucleus possibly through the interference of neutrino particles via the Trivedi Effect^®-Consciousness Energy Healing Treatment. The neutrinos have the ability to interact with protons and neutrons in the nucleus, which indicated a close relationship between neutrino and the isotope formation 233839. The isotopic abundance ratios 2H/1H or 13C/12C or 17O/16O would influence the atomic bond vibration of treated ascorbic acid 44. The increased isotopic abundance ratio of the treated ascorbic acid may increase the intra-atomic bond strength, its physical stability. This novel ascorbic acid after the Trivedi Effect^®-Consciousness Energy Healing Treatment would be very useful to design more efficacious pharmaceutical formulations against scurvy, obesity, cardiovascular diseases, hypertension, rheumatoid arthritis, Alzheimer's disease, cancer, etc.

Conclusion

Conclusions

The impact of the Trivedi Effect^®-Consciousness Energy Healing Treatment on ascorbic acid was identified as significant in the isotopic abundance ratio and chromatographic peak area. The control and Consciousness Energy Healing Treated samples showed a major chromatographic peak at retention time (R_t) 1.8 minutes exhibited the deprotonated molecular ion peak at m/z 175 ^M^- in the mass spectra. The peak area of the Consciousness Energy Healing Treated ascorbic acid was significantly increased by 8.81% compared to the control sample. The LC-MS based isotopic abundance ratio of P_M+1/P_M (2H/1H or 13C/12C or 17O/16O) in the Consciousness Energy Healing Treated ascorbic acid was significantly increased by 23.22% compared with the control sample. Thus,13C, 2H, and17O contributions from (C₆H₇O₆)^- to m/z 176 in the Consciousness Energy Healing Treated ascorbic acid were significantly increased compared with the control sample. The increased isotopic composition of the Consciousness Energy Healing Treated ascorbic acid might have altered the neutron to proton ratio in the nucleus. The changes in isotopic abundance could be due to changes in nuclei possibly through the interference of neutrino particles via the Trivedi Effect^®-Consciousness Energy Healing Treatment. The increased isotopic abundance ratio and peak area of the Consciousness Energy Healing Treated ascorbic acid may increase the intra-atomic bond strength and its stability. This novel ascorbic acid after the Trivedi Effect^®-Consciousness Energy Healing Treatment would be very useful to design more efficacious pharmaceutical formulations against scurvy, obesity, cardiovascular diseases, hypertension, rheumatoid arthritis, Alzheimer's disease, cancer, etc.

Journal of Advances in Nanotechnology