Study of the Energy of Consciousness Healing Treatment on Physical, Structural, Thermal, and Behavioral Properties of Zinc Chloride

Zinc chloride is an important pharmaceutical/nutraceutical mineral for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect-Energy of Consciousness Healing Treatment (Biofield Energy Treatment) on physical, structural, thermal, and behavioral properties of zinc chloride using PXRD, PSD, FT-IR, UV-vis, and DSC analysis. Zinc chloride was divided into two parts – one part was control, while another part was treated with The Trivedi Effect remotely by seven renowned Biofield Energy Healers and defined as The Trivedi Effect treated sample. A significant alteration of the crystallite size and relative intensities of the PXRD peaks were observed in The Trivedi Effect treated sample with respect to the control sample. A sharp peak at 2θ equal to 51.45°, which was observed in the control sample was disappeared in the treated sample. The average crystallite size of the treated sample was decreased by 2.13% compared with the control sample. The size of the particles at d10, d50, and d90 was decreased by 3.36%, 7.97%, and 11.51%, respectively in the treated sample compared with the control sample. The surface area of the treated sample was significantly increased by 5.69% compared with the control sample. The FT-IR spectroscopic analysis revealed that Zn-Cl stretching in the control was 646 cm, whereas it was significantly shifted downward to 505 cmin the treated sample. The UV-vis analysis exhibited that wavelength of the maximum absorbance (λmax) of both the control and treated samples was at 196.4 nm. The DSC analysis exhibited that the melting and decomposition temperature were increased by 0.93% and 0.56%, respectively in the treated zinc chloride compared to the control sample. The enthalpy of fusion of the treated sample (338.27 J/g) was increased significantly by 33.84% compared with the control sample (252.75 J/g). The enthalpy of decomposition of the treated sample was increased by 3.78% compared with the control sample. The current study anticipated that The Trivedi Effect-Energy of Consciousness Healing Treatment might lead to produce a new polymorphic form of zinc chloride, which would be more soluble, bioavailable, and thermally stable compared with the untreated compound. The treated sample could be more stable during manufacturing, delivery or storage conditions than the untreated sample. Hence, the treated zinc chloride would be very useful to design better nutraceutical/pharmaceutical formulations that might offer better therapeutic response against inflammatory diseases, immunological disorders, aging, stress, cancer, etc.


Introduction
Zinc is an essential element in human and animal nutrition for its broad spectrum of biological activities. It plays an important role in catalytic, structural or regulation in enzymes in our body, which involved in the nucleic acid and protein metabolism as well as for the production of energy. Zinc also maintains the structural integrity of the biological membranes resulting in their protection against oxidative injury [1][2]. As a metallotherapeutic agent, zinc possesses various pharmacological activities include fertility enhancing, retino-protective, and putative antiviral activities [3]. Zinc is also used in the treatment of Wilson's disease [4]. It has also immunomodulatory and antioxidant activities [1]. Zinc is also used to prevent the development of several deficiency symptoms include parakeratosis, hypogeusia, anorexia, dysosmia, geophagia, hypogonadism, growth retardation, etc. [5][6][7]. Recently, it has been reported that zinc had increased the leukocyte count and phagocytic index, which potentiate the immunomodulatory effect along with Glycyrrhiza glabra [8]. Zinc chloride (ZnCl 2 ) is hygroscopic inorganic salt and highly soluble in water [9]. It is sometimes used in pharmaceutical industry as drug and diagnostic agent due to its powerful astringent and mild antiseptic properties [10]. It is also used in mouth-wash and deodorant preparations. Zinc chloride is used in as dentin desensitizer i.e. reduces the sensitivity of the teeth to heat and cold. The other applications of zinc chloride include protein precipitation and insulin preparation [10,11]. Literature reported that zinc chloride has been used in the treatment of cancer as a destructive agent [12] and in the pluripotency maintenance of mouse embryonic stem cell through the regulation of STAT3 signaling pathway [13]. It can be used for the palliative treatment of canine and malignant skin wounds [14]. It has the opposite effect on locomotor behavior of rats [15]. Therefore, zinc chloride was considered as one of the component in the novel proprietary herbomineral formulation for the source of zinc ion. This herbomineral formulation is designed as nutraceutical supplement, and can be used for the prevention and treatment of various human disorders.
An élan vital or vital force which contributes the 'life' is preserved by every living organisms and known 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 is considered as the Biofield Energy. The Biofield Energy is infinite, paradimensional and dynamic electromagnetic field surrounding the human body. It can freely flow between the human and environment that leads to the continuous movement or matter of energy [16,17]. Thus, a human has the ability to harness energy from the earth, the "universal energy field" and transmit it to any living or nonliving object(s) around the globe. The objects always receive the energy and respond in a useful way. This process is known as Biofield Energy Healing Treatment [18,19]. Biofield (Putative Energy Fields) based Energy Therapies are used worldwide to promote health and healing. The National Center of Complementary and Integrative Health (NCCIH) has been 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) [20]. Biofield Energy Treatment (The Trivedi Effect ® ) has been reported in numerous peer-reviewed science journals with significant outcomes in many scientific fields such as cancer research [21]; altered antimicrobial sensitivity of pathogenic microbes in microbiology [22][23][24], biotechnology [25,26], genetics [27,28]; changing the structure of the atom in relation to various metals, ceramics, polymers and chemicals in materials science [29,30], altered physical and chemical properties of pharmaceuticals [31,32], nutraceuticals [33,34], organic compounds [35][36][37], and improved overall growth and yield of plants in agricultural science [38,39]. The physicochemical properties such as particle size, crystalline structure, crystallite size, surface area, and thermal properties of a drug have a vital role in bioavailability as well as stability of the drug during processing, formulation, storage, and packaging [40,41]. The particle size, specific surface area, crystalline nature, chemical and thermal behavior of an atom/ion might be altered by the Biofield Energy Healing Treatment (The Trivedi Effect ® ) through possible mediation of neutrinos [42]. By considering all these aspects, powder X-ray diffraction (PXRD), particle size distribution analysis (PSD), Fourier transform infrared (FT-IR) spectrometry, ultraviolet-visible (UV-vis) spectroscopy, and differential scanning calorimetry (DSC) analytical techniques were conducted in this study for the characterization of physical, structural, and thermal properties of the Biofield Energy Treated and untreated zinc chloride.

Chemicals and Reagents
Zinc chloride was procured from Tokyo Chemical Industry Co., Ltd. (TCI), Japan. All other chemicals used in the experiment were of analytical grade available in India.

The Trivedi Effect ® -Energy of Consciousness Healing Treatment Strategies
Zinc chloride was one of the components of the new proprietary herbomineral formulation, developed by our research team and it 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 zinc chloride sample. The second part of the test compound was treated with The Trivedi Effect ® -Energy of Consciousness Healing Treatment (Biofield Energy Treatment) by a group of seven renowned Biofield Energy Healers remotely and was denoted as Biofield Energy Treated or The Trivedi Effect ® sample. Six Biofield Energy Healers were remotely located in the U.S.A. and one of which was remotely located in Canada, while the test compound was located in the research laboratory of GVK Biosciences Pvt. Ltd., Hyderabad, India. This Biofield Energy Treatment was provided for 5 minutes through Healer's Unique Energy Transmission process remotely to the test compound under the 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" healers for 5 minutes, under the same laboratory conditions. The sham healer did not have any knowledge about the Biofield Energy Treatment. After that, the Biofield Energy Treated and untreated samples were kept in similar sealed conditions and characterized thoroughly by PXRD, PSD, FT-IR, UV-visible spectroscopy, and DSC analysis.

Powder X-Ray Diffraction (PXRD) Analysis
The PXRD analysis was accomplished on PANalytical X'pert Pro powder X-ray diffractometer system. The X-ray of wavelength 1.54056 Å was used. The data was collected in the form of a chart of the Bragg angle (2θ) vs. intensity, and a detailed table containing information on peak intensity counts, d value (Å), relative intensity (%), full width half maximum (FWHM) (θ°). From the PXRD results, the crystallite size (G) was calculated using X'pert data collector and X'pert high score plus processing software. The crystallite size (G) was calculated from the Scherrer equation [43,44]. The method was based on the width of the diffraction patterns obtained in the X-ray reflected the crystalline region. The crystallite size (G) was calculated by using the following formula (1): Where, k is the equipment constant (0.5), λ is the X-ray wavelength (0.154 nm); b in radians is the full-width at half of the peaks and θ the corresponding Bragg angle.
Percent change in crystallite size (G) was calculated using following equation (2): Where, G Control and G Treated are the crystallite size of the control and Biofield Energy Treated samples, respectively.
A total of ~500.21 mg of the control and treated samples individually were used for the analysis and prepared by back loading technique using the sample preparation kit. The sample was spread on the holder ring in sufficient quantity to fill the ring cavity. It was then pressed down using powder press block and scrap the powder that was in surplus using a glass slide to get densely packed specimen. Consequently, the bottom plate was placed onto the holder ring and clamp in position. The sample holder was then removed from the sample preparation table by turning it upside down. A smooth surface of sample was obtained to ensure optimum results.

Particle Size Distribution (PSD) Analysis
The average particle size and particle size distribution were analyzed using Malvern Mastersizer 2000, UK with a detection range between 0.01 µm to 3000 µm. The sample unit was filled with the dispersant medium and operated the stirrer at 2500 rpm. Alignment of the optics was done and the background measurement was taken. After the background measurement, the sample was added into the sample unit with constant monitoring the obscuration and stopped the addition of sample when the obscuration reached in between 15% to 20%. When the obscuration was stable, the measurement was taken twice and the average of two measurements was taken. The average histogram of the two measurements was recorded. The print out of the average histogram of the two measurements was documented. Along with histogram, the data was presented in table format which include particle size (µm). Also, the values at below 10% level (d 10 ), 50% level (d 50 ), and 90% level (d 90 ) were calculated from the histogram and the calculations such as surface area (m 2 /g) were done by using software Mastersizer 2000.
Percent change in particle size (d) for at below 10% level (d 10 ), 50% level (d 50 ), and 90% level (d 90 ) was calculated using following equation (3): Where, d Control and d Treated are the particle size (µm) for at below 10% level (d 10 ), 50% level (d 50 ), and 90% level (d 90 ) of the control and treated samples, respectively.
Percent change in surface area (S) was calculated using following equation (4): Where, S Control and S Treated are the surface area of the control and treated samples, respectively.

Fourier Transform Infrared (FT-IR) Spectroscopy
FT-IR spectroscopy of zinc chloride was performed on Spectrum two (Perkin Elmer, USA) Fourier transform infrared spectrometer with the frequency array of 400-4000 cm -1 by using pressed KBr disk technique.

Ultraviolet-Visible Spectroscopy (UV-Vis) Analysis
The UV-Vis spectral analysis was carried out using Shimadzu UV-2450 with UV Probe, Japan. The spectrum was recorded using 1 cm quartz cell that has a slit width of 1.0 nm. The wavelength range chosen for recording the spectra was 190-800 nm. The absorbance spectra (in the range of 0.2 to 0.9) and wavelength of maximum absorbance (λ max ) were recorded.

Differential Scanning Calorimetry (DSC)
Analysis was performed using the DSC Q20 (TA Instruments, USA) differential scanning calorimeter. A total of ~7.50 mg sample was weighed and sealed in aluminum pans and equilibrated at 25°C and heated up to 450ºC at the heating rate of 10°C/min under nitrogen gas as purge atmosphere with flow rate of 50 mL/min. The value for onset, endset, peak temperature, peak height (mJ or mW), peak area, and change in heat (J/g) for each peak were recorded.
Percent change in the decomposition temperature (T) was calculated using following equation (5): % change in melting point ) ) ) 100 (5) Where, T Control and T Treated are the decomposition temperature of the control and treated samples, respectively.
Percent change in the enthalpy of decomposition (∆H) was calculated using following equation (6): Where, ∆H Control and ∆H Treated are the enthalpy of decomposition of the control and treated samples, respectively.

Powder X-Ray Diffraction (PXRD) Analysis
The PXRD diffractograms of both the control and treated zinc chloride are shown in the Figure 1. The diffractograms displayed very sharp and intense peaks in their respective diffractogram indicating the crystalline nature of zinc chloride. PXRD data such as Bragg angle (2θ), relative intensity (%), full width half maximum (FWHM) (θ°), and crystallite size (G) for the control and Biofield Energy Treated zinc chloride are presented in the    (Figure 1) was observed in the control sample, whereas this peak was absent in the treated sample. In addition, the XRD peaks at 2θ equal to 70.67° and 72.28° (Table 1, entry 16 and 17) in the control sample were shifted to 72.20° and 73.89° (Table 1, entry 16 and 17) in the treated sample. These results indicated the relative intensities of the XRD peaks (Table  1) in the treated sample were significantly altered as compared to the control sample. It has been found in the scientific literature that XRD relative intensity of each diffraction face on the crystalline compound changes according to the crystal morphology [45]. Thus, it is presumed that the energy transferred through the Biofield Energy Healing Treatment probably altered the shape and size of molecules, which might be responsible for an alteration in the relative intensities and crystallite size in the treated sample as compared to the control sample.
Raza et al. reported that changes in XRD pattern provide the proof of polymorphic transitions [46,47]. Crystal habit, size, and even polymorphic form of a drug have a significant effect on the drug solubility, dissolution, and bioavailability. It has been reported in the literature that alteration in the crystal morphology had the significant impact on in vitro dissolution rate, with prospective for improving bioavailability [41]. Hence, PXRD study revealed that Biofield Energy Healing Treatment might introduced a new polymorphic form of zinc chloride, which could have improved bioavailability compared with the control sample.

Particle Size Analysis
The particle size data (d 10 , d 50 , and d 90 ) of both the control and treated zinc chloride were investigated and the results are presented in the Table 2. *denotes the percentage change in the particle size data (d10, d50, and d90) and surface area of the Biofield Energy Treated sample with respect to the control sample.

It was evident that Biofield Energy Treatment (The Trivedi
Effect ® treated) has the ability to reduce the particle size (d 10 , d 50 , and d 90 ) of zinc chloride with respect to the control sample. The size of the particles at below 10% level (d 10 ), 50% level (d 50 ), and 90% level (d 90 ) was decreased by 3.06%, 7.97%, and 11.51%, respectively in the Biofield Energy Treated sample compared to the control sample. Particle size as well as the surface area have a major effect on dissolution of a compound in a solvent, because the surface energy influenced by surface area and chemical affinity is the driving factor for dissolution efficiency [48,49]. The surface area for both the control and Biofield Energy Treated zinc chloride samples was analyzed and the results are presented in Table 2. The control sample showed surface area of 2.99 m 2 /g and it was increased to 3.16 m 2 /g in the Biofield Energy Treated zinc chloride. The result showed 5.69% increase in surface area of the Biofield Energy Treated sample with respect to the control sample.
Poorly crystallized compounds possess more surface area and higher exchange capacities than well-crystallized compounds [50]. In addition, the variation of the crystal morphology in the Biofield Energy Treated sample, which was well-supported from PXRD data, may cause to alter the surface area of the Biofield Energy Treated zinc chloride compared with the control sample. It has been well established that the particle size, shape and surface area of pharmaceutical compounds have an important impact on solubility, dissolution and in vivo bioavailability, as well as supports the designing of new drug delivery systems [51,52]. Reducing particle size and higher surface area would enhance the solubility of the solid particles, and consequently would increase the dissolution rate and bioavailability [53]. Thus, it is assumed that the Biofield Energy Treated zinc chloride might be dissolved and absorbed at a faster rate and may possibly have more bioavailability than untreated zinc chloride.

Fourier Transform Infrared (FT-IR) Spectroscopy
The FT-IR spectra of the control and Biofield Energy Treated samples of zinc chloride are presented in the Figure  2. The characteristic peak were observed at 3496, 1616, 1403, and 646 cm -1 in the control FT-IR spectrum (Figure 2). On the other hand, The FT-IR spectrum of the Biofield Energy Treated zinc chloride (Figure 2) showed the notable absorption bands at 3590 cm -1 , 3519 cm -1 , 1931 cm -1 , 1608 cm -1 , 993 cm -1 , and 505 cm -1 . The strong and broad absorption band centered at 3496 cm -1 and the H-O-H bending motion at 1616 cm -1 that were due to the presence of the lattice water were found in the control FT-IR spectrum ( Figure 2). Consequently, the peaks for the lattice water were observed in the spectrum of the Biofield Energy Treated zinc chloride ( Figure 2) at 3590, 3519, and 1608 cm -1 . From the literature, it has been found that metal stretching absorption band for inorganic materials was found in the 750-100 cm -1 region [54]. In control spectrum, metal-halogen (Zn-Cl) stretching was shown at 646 cm -1 , whereas it was shifted downward in the Biofield Energy Treated sample at 505 cm -1 . Overall, FT-IR results suggest that there may be a significant impact of Biofield Energy Treatment on zinc chloride at the atomic level to reduce the force constant of Zn-Cl bonds.

Ultraviolet-Visible Spectroscopy (UV-Vis) Analysis
The UV-visible spectra of both the control and treated zinc chloride are presented in the Figure 3. The wavelength of the most intense UV-Vis absorbance (λ max ) of the both the control and Biofield Energy Treated samples were observed in their UV-Vis spectra (Figure 3) at 196.0 nm with a minor shift of absorbance maxima from 2.0317 (control) to 1.9653 in the treated sample. The UV absorbance occurs due to the diverse type of energy transitions from the singlet to the singlet excited state such as σ→σ * , n→π * , and π→π * . These type of electronic transitions are occurred when the difference in energy between the lowest unoccupied molecular orbital (LUMO) and the highest occupied molecular orbital (HOMO) is significantly higher than the activation energy of the compound [55]. The UV-vis spectroscopic analysis of zinc chloride displayed no change in the λ max of the treated sample as compared to the control sample. Thus, the structural configuration or activation energy of the treated sample was not changed as compared to the control sample.

Differential Scanning Calorimetry (DSC) Analysis
The DSC thermograms of the control and treated zinc chloride are presented in the Figure 4. The melting/decomposition temperature and enthalpy of fusion/decomposition of the control and treated zinc chloride are presented in the Table 3. The melting/fusion temperature was increased by 0.93% in the treated zinc chloride (315.12°C) as compared to the control sample (312.23°C). The onset and endset melting temperatures of the treated sample were also increased by 2.50% and 0.62%, respectively compared with the control sample. The enthalpy of fusion (∆H fusion ) of the control and treated zinc chloride were 252.75 J/g and 338.27 J/g, respectively. The ∆H fusion of the treated sample was increased significantly by 33.84% compared with the control sample. Similarly, the decomposition temperature of the treated zinc chloride (385.06°C) was increased by 0.56% compared to the control sample (382.90°C). The onset and endset decomposition temperatures of the treated sample were also increased by 0.19% and 0.50%, respectively compared with the control sample. The enthalpy of decomposition (∆H decomposition ) of the control and treated zinc chloride were 1173.25 J/g and 1217.56 J/g, respectively. The ∆H decomposition of the treated sample was increased by 3.78% compared with the control sample. It is anticipated that Biofield Energy Treatment might enhance the internal energy in zinc chloride that leads to increase in the melting/decomposition temperature and the enthalpy of fusion/decomposition as well as thermal stability of the Biofield Energy Treated sample. The enhanced thermal stability of a drug is an advantage in the maintenance of the product in long term storage condition throughout the shelflife for any pharmaceutical formulations [56].