Evaluation of the Physicochemical, Structural, Thermal, and Behavioral Properties of the Energy of Consciousness Healing Treated Zinc Chloride

Zinc chloride is a source of zinc used in various pharmaceutical/nutraceutical formulations. 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, UVvis, 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 twenty 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 was observed in The Trivedi Effect treated sample compared with the control sample. The average crystallite size of the treated sample was significantly increased by 23.18% compared with the control sample. The particle size values at d10, d50, and d90 values were significantly decreased by 3.70%, 4.13%, and 6.13%, respectively in the treated sample compared with the control sample. Therefore, the surface area of the treated sample was increased by 4.21% compared with the control sample. The FT-IR spectroscopic analysis revealed that Zn-Cl stretching in the control sample was found at 512 cm, whereas it was significantly shifted upward to 520 cm in the treated sample. The UV-vis analysis exhibited that wavelength of the maximum absorbance (λmax) of the control and treated samples were at 197.6 nm and 197.1 nm, respectively. The DSC analysis exhibited that the melting temperature was decreased by 0.22%, while decomposition temperature was increased by 2.56% in the treated sample compared to the control sample. The latent heat of fusion of the treated sample (320.44 J/g) was significantly decreased by 16.70% compared with the control sample (284.67 J/g). Similarly, the enthalpy of decomposition of the treated sample (952.53 J/g) was significantly increased by 122.61% compared with the control sample (427.90 J/g). Thus, the results indicated that the thermal stability of the treated zinc chloride was improved compared with the control sample. The current study anticipated that The Trivedi Effect Energy of Consciousness Healing Treatment might lead to produce a thermally stable new polymorphic form of zinc chloride, which would be more soluble and bioavailable compared with the untreated compound. Hence, the treated zinc chloride would be very useful to design better nutraceutical/pharmaceutical formulations that might offer better therapeutic response against American Journal of Bioscience and Bioengineering 2017; 5(2): 65-74 66 inflammatory diseases, immunological disorders, aging, stress, cancer, etc.


Introduction
Zinc is an essential element in human body due to its broad spectrum of physiological functions. It plays an important role in catalytic, structural or regulation in enzymes in the 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, retinoprotective, 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 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 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 also 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 components 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 [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. Properties of the Energy of Consciousness Healing Treated Zinc Chloride

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 twenty renowned Biofield Energy Healers remotely and was denoted as Biofield Energy Treated or The Trivedi Effect ® sample. Thirteen Biofield Energy Healers were remotely located in the U.S.A., five were located in Canada, and two were located in Australia, 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: % change in crystallite size = × 100 (2) Where, G Control and G Treated are the crystallite size of the control and Biofield Energy Treated samples, respectively.
A total of ~500 mg of the control and Biofield Energy 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 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 Biofield Energy 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 Biofield Energy 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 ~5 mg sample was weighed and sealed in aluminum pans and equilibrated at 25°C and heated up to 450°C 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 melting/decomposition temperature (T) was calculated using following equation 5: Where, ∆H Control and ∆H Treated are the enthalpy of decomposition of the control and Biofield Energy Treated samples, respectively.

Powder X-ray Diffraction (PXRD) Analysis
The PXRD diffractograms of both the control and Biofield Energy Treated zinc chloride are presented in Figure 1. Both the samples 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 Table 1. The crystallite size was calculated with the help of Scherrer equation [43,44]. The crystallite size of the control and Biofield Energy Treated samples were found to be similar at 2θ equal to nearly 48.6°, 56.2°, and 70.5° (Table 1, entry 6, 9, and 13   Scientific literature reported that the changes in the XRD patterns, such as crystallite size and relative intensities, indicated the modification of the morphology of the crystal as well as the proof of polymorphic transition [45][46][47]. As the crystal morphology of the Biofield Energy Treated sample was altered compared with the control sample, the Biofield Energy Treated sample might be a new polymorphic form of zinc chloride. The crystal pattern, size and even polymorphic form of a pharmaceutical play important roles in drug solubility, dissolution and bioavailability. It has been reported in the literature that the alteration in crystal morphology has significant impact on the in vitro dissolution rate, which is related with the bioavailability of orally administered pharmaceutical/nutraceutical [41]. So, it can be concluded that the Biofield Energy Healing Treatment might be a very useful method for enhancing the bioavailability of zinc chloride.

Particle Size Distribution (PSD) Analysis
The particle size values (d 10 , d 50 , and d 90 ) and the average surface area of the control and Biofield Energy Treated zinc chloride were investigated and the results are presented in the Table 2. It was evident that the Biofield Energy Treatment 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 ) were decreased by 3.70%, 4.13%, and 6.13%, respectively in the Biofield Energy Treated sample compared to the control sample. * 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.
The particle size and surface area have a major effect on dissolution of a compound in a solvent, because the surface energy influenced by the 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 were analyzed and the results are presented in Table 2. The control sample showed surface area of 3.09 m 2 /g and it was increased to 3.22 m 2 /g in the Biofield Energy Treated zinc chloride. The surface area of the Biofield Energy Treated sample increased by 4.21% compared with 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 the 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 peaks were observed at 3904, 3583, 3518, 1931, 1609, 986, and 512 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 showed the notable absorption bands at 3905, 3583, 3519, 1931, 1608, 985, and 520 cm -1 (Figure 2). The strong and broad absorption band centered near 3583 cm -1 and the H-O-H bending motion at 1608 cm -1 that were due to the presence of the lattice water were found in the FT-IR spectrum of both the control and Biofield Energy Treated samples (Figure 2). Literature suggested that the metal-halide stretching absorption band for inorganic materials generally found in the 750-100 cm -1 region [54]. In control spectrum, metal-halogen (Zn-Cl) stretching was found at 512 cm -1 , whereas it was shifted upward in the Biofield Energy Treated sample at 520 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 increase the force constant of Zn-Cl bonds. This result also supported with the PXRD data that Biofield Energy Treated sample might be new polymorphic form of zinc chloride.

Ultraviolet-Visible Spectroscopy (UV-Vis) Analysis
The UV-visible spectra of both the control and Biofield Energy Treated zinc chloride are presented in the Figure 3. The spectra of control and Biofield Energy Treated sample showed the maximum absorbance (λ max ) at 197.6 nm and 197.1 nm, respectively, along with similar shift of absorbance maxima from 1.4723 in control and 1.4727 in Biofield Energy 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 Biofield Energy Treated sample compared to the control sample. Thus, the structural configuration or activation energy of the Biofield Energy Treated sample was not changed compared with the control sample. Properties of the Energy of Consciousness Healing Treated Zinc Chloride

Differential Scanning Calorimetry (DSC) Analysis
The DSC thermograms of the control and Biofield Energy Treated zinc chloride are presented in the Figure 4. The melting/decomposition temperature and enthalpy of fusion/decomposition of the control and Biofield Energy Treated zinc chloride are presented in the Table 3. The melting/fusion temperature was slightly decreased by 0.22% in the Biofield Energy Treated zinc chloride (311.01°C) compared with the control sample (311.71°C). The onset melting temperature was increased by 1.47% and the endset temperature was decreased slightly by 0.26% in the Biofield Energy Treated sample compared with the control sample. The latent heat of fusion or enthalpy of fusion (∆H fusion ) of the control and Biofield Energy Treated zinc chloride was 384.67 J/g and 320.44 J/g, respectively. The ∆H fusion of the Biofield Energy Treated sample was significantly decreased by 16.70% compared with the control sample. Similarly, the decomposition temperature of the Biofield Energy Treated zinc chloride (384.29°C) was increased by 2.56% compared to the control sample (392.59C). The onset and endset decomposition temperature was increased by 1.95% and 2.83% in the Biofield Energy Treated sample compared with the control sample. The enthalpy of decomposition (∆H decomposition ) of the control and Biofield Energy Treated zinc chloride were 427.90 J/g and 952.53 J/g, respectively. The ∆H decomposition of the Biofield Energy Treated sample was significantly increased by 122.61% compared with the control sample. Overall, results indicated that the zinc chloride need more energy in the form of latent heat to undergo the process of decomposition after Biofield Energy Treatment. It is anticipated that Biofield Energy Treatment might enhance the internal energy in zinc chloride that leads to increase of the decomposition temperature and the enthalpy of decomposition and ultimately to enhance the overall thermal stability of the Biofield Energy Treated sample significantly compared with the untreated sample. The enhanced thermal stability of a drug is an advantage in the maintenance of the product in long term storage condition throughout the shelf-life for any pharmaceutical formulations [56].

Conclusions
The current study revealed that The Trivedi Effect ® -Energy of Consciousness Healing Treatment had the significant effects on the physical, structural, thermal, and behavioral properties of zinc chloride. The PXRD data displayed the significant alteration of the crystallite size and relative intensities of the PXRD peaks in The Trivedi Effect ® treated sample compared with the control sample. The average crystallite size of the treated sample was significantly increased by 23.18% compared with the control sample. The particle size values at d 10 , d 50 , and d 90 were significantly decreased by 3.70%, 4.13%, and 6.13%, respectively in the treated sample compared with the control sample. Therefore, the surface area of the treated sample was increased by 4.21% compared with the control sample. The FT-IR spectroscopic analysis revealed that Zn-Cl stretching in the control sample was 512 cm -1 , whereas it was significantly shifted upward to 520 cm -1 in the treated sample. The DSC analysis exhibited that the melting temperature was decreased by 0.22%, while decomposition temperature was increased by 2.56% in the treated sample compared to the control sample. The latent heat of fusion of the treated sample was significantly decreased by 16.70% compared with the control sample. Similarly, the enthalpy of decomposition of the treated sample was significantly increased by 122.61% compared with the control sample. The results indicated that the thermal stability of the treated zinc chloride was improved compared with the control sample. Thus, the treated sample could be more stable during manufacturing, delivery or storage conditions than the untreated sample. Briefly, The Trivedi Effect ® -Energy of Consciousness Healing Treatment might introduce a new thermally stable polymorphic form of zinc chloride that could be more soluble and bioavailable compared with the untreated compound. Hence, Biofield Energy Treated zinc chloride would be very useful to design better nutraceutical and/or pharmaceutical formulations that might offer better therapeutic response against inflammatory diseases,