A Comprehensive Analytical Evaluation of the Trivedi Effect ®-Energy of Consciousness Healing Treatment on the Physical , Structural , and Thermal Properties of Zinc Chloride

Zinc chloride has an importance in pharmaceutical and nutraceutical industries 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, and thermal properties of zinc chloride using PXRD, PSD, FT-IR, UV-vis, TGA, and DSC analysis. Zinc chloride was divided into two parts – one part was control, while other part was treated with The Trivedi Effect remotely by eighteen renowned Biofield Energy Healers and defined as The Trivedi Effect Treated sample. A significant change of the crystallite size and relative intensities of the PXRD peaks was observed in The Trivedi Effect treated sample with respect to the control sample. The average crystallite size of the treated sample was significantly increased by 41.22% compared with the control sample. The size of the particles at d10, d50, and d90 was increased by 8.33%, 6.27%, and 1.83%, respectively in the treated sample compared with the control sample. The surface area of the treated sample was decreased by 5.90% compared with the control sample. The FT-IR spectroscopic analysis revealed that Zn-Cl stretching in the control and treated sample were at 511 and 507 cm, respectively. The UV-vis analysis exhibited that wavelength of the maximum absorbance of both the control and treated samples were at 196.4 and 196.3 nm, respectively. The TGA analysis exhibited that weight loss of the treated sample was reduced by 13.98% and 2.43% in the 1 and 2 steps of degradation, respectively. Consequently, the DSC analysis revealed that the maximum thermal decomposition temperature of the treated sample (489.06°C) was significantly increased by 15.75% compared with the control sample (422.50°C). Moreover, a significant increase in the enthalpy of decomposition was noticed in the treated sample (∆Hdecomp = 131.40 J/g) by 137.10% compared with the control sample (∆Hdecomp = 55.42 J/g). The current study anticipated that The Trivedi Effect Treatment might lead to produce a new polymorphic form of zinc chloride, which could have better powder flowability and thermal stability. 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 and/or 8 Mahendra Kumar Trivedi et al.: A Comprehensive Analytical Evaluation of the Trivedi Effect Energy of Consciousness Healing Treatment on the Physical, Structural, and Thermal Properties of Zinc Chloride pharmaceutical formulations that might offer better therapeutic response against inflammatory diseases, immunological disorders, aging, stress, cancer, etc.


Introduction
Zinc is a vital mineral element in human and animal nutrition with a wide array of biological activities. It plays an important roles in catalytic, structural or regulation in greater than 200 zinc metalloenzymes identified in the biological systems [1,2]. These enzymes are 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]. 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 useful for the palliative treatment of canine and malignant skin wounds [14]. It has 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]. So, 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, etc. 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, thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC) analytical 9 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 TCI, Japan. All other chemicals used in the experiment were of analytical grade available in India.

The Trivedi Effect ® Energy of Consciousness 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 was treated with The Trivedi Effect ® -Energy of Consciousness Healing Treatment (Biofield Energy Treatment) by the renowned Biofield Energy Healers and defined as Biofield Energy Treated sample, while the second part of the test compound did not receive any sort of treatment and defined as untreated or control zinc chloride sample. This Biofield Energy Treatment was provided by the group of eighteen renowned Biofield Energy Healers who participated in this study and performed the Biofield Energy Treatment remotely. Eleven Biofield Energy Healers were remotely located in the U. S. A., four remotely located in Canada, two remotely located in Finland, and one of which was remotely located in Albania, 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, TGA, 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 equation 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 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% and 20%. When the obscuration was stable, the measurement was taken twice and the average was taken of two measurements. The average histogram of the two measurements was recorded. 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, 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.

Thermal Gravimetric Analysis (TGA)
The TGA analysis was performed using Instrument TGA Q50 (TA Instruments, USA) at a heating rate of 10ºC/min from room temperature i.e. 30°C to 900°C under nitrogen atmosphere. A total of ~ 10 mg of sample was used for the analysis and was taken on the platinum pan. In TGA, the weight loss for each step was recorded in grams as well as in percent loss with respect to initial weight. Also, the onset, endset, and peak temperature for each step were recorded in TGA. Percent change in weight loss (W) was calculated using following equation 5: Where, W Control and W Treated are the weight loss of the control and Biofield Energy Treated samples, respectively.

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 600ºC 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 6: Where, T Control and T Treated are the decomposition temperature of the control and Biofield Energy Treated samples, respectively.
Percent change in the enthalpy of decomposition (∆H) was calculated using following equation 7: 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 shown in Figure 1. Very sharp and intense peaks in both the diffractograms 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 Table 1. The crystallite size was calculated with the help of Scherrer equation [43,44]  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 be introduced a new polymorphic form of zinc chloride, which could have improved bioavailability as compared with the control sample.

Particle Size Distribution (PSD) Analysis
The particle size data (d 10 , d 50 , and d 90 ) of both the control and Biofield Energy Treated zinc chloride was investigated and the results are presented in Table 2. It was evident that Biofield Energy Treatment has the ability to enhance 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 increased by 8.33%, 6.27%, and 1.83%, respectively in the Biofield Energy Treated sample as 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 3.05 m 2 /g and it was decreased to 2.87 m 2 /g in the Biofield Energy Treated zinc chloride. The result showed 5.9% decrease in surface area of the Biofield Energy Treated sample with respect to the control sample. Kale et al. [50] reported that introduction of the external force leads to the transform the fine particles into larger particles. The various reasons to increase the particle size are enhanced the powder flowability, improved product shape and appearance [50]. It is presumed that the Biofield Energy Healing Treatment (i.e. consider as external force) might improve the powder flowability, shape and appearance of zinc chloride. XRD data also supported that the Biofield Energy Healing Treatment could alter the size, shape, and appearance of 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 Figure 2. The peaks found in the control IR spectrum were at 3585, 3518, 1607, and 511 cm -1 , whereas the Biofield Energy Treated zinc chloride showed the peaks at 3588, 3518, 1607, and 507 cm -1 . The strong and broad absorption band centered at 3585 and 3588 cm -1 , while the H-O-H bending motion was noticed at 1607 cm -1 in the FT-IR spectra indicated the presence of the lattice water in both the control and Biofield Energy Treated zinc chloride samples. The fingerprint region of the Biofield Energy Treated and control samples was remained same. From the literature, it has been found that metal stretching absorption band for inorganic materials was found in the 750-100 cm -1 region [51]. In control spectrum, metal-halogen (Zn-Cl) stretching was shown at 511 cm -1 , whereas it was displayed in the Biofield Energy Treated sample at 507 cm -1 . Overall, IR peaks of the Biofield Energy Treated zinc chloride did not show any significant changes with respect to the control sample.

Ultraviolet-Visible Spectroscopy (UV-Vis) Analysis
The UV-visible spectra of the both control and Biofield Energy Treated zinc chloride are presented in Figure 3. The UV-visible spectra exhibited that the maximum absorbance (λ max ) of both the control and Biofield Energy Treated samples were at 196.4 and 196.3 nm, respectively with a minor shift of absorbance maxima from 1.9984 to 1.9784 in the control and Biofield Energy Treated samples, respectively. The UV absorbance occurs due to the different 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 happened 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 [52]. The UV-vis spectroscopic analysis of zinc chloride showed no change in the λ max of the Biofield Energy Treated sample as compared to the control sample. Thus, the structural configuration or activation energy of the Biofield Energy Treated sample was not changed as compared to the control sample.

Thermal Gravimetric Analysis (TGA)
The TGA thermograms of both the control and Biofield Energy Treated samples are presented in Figure 4. The TGA thermogram of both the control and Biofield Energy Treated samples exhibited two steps thermal degradation and mentioned in Table 3. The pattern of the thermal degradation of the control zinc chloride was nearly matched with the reported data [53]. The weight loss of the 1 st step (8.37%) and 2 nd step (85.87%) degradation in the Biofield Energy Treated zinc chloride was significantly reduced by 13.98% and 2.43%, respectively as compared with the control sample (1 st step = 9.73% and 2 nd step = 88.01%).  The total weight loss of the Biofield Energy Treated sample (94.24%) was significantly reduced by 3.58% compared with the control sample (97.74). Overall, the TGA study revealed that the Biofield Energy Treated zinc chloride was thermally more stable as compared with the control sample.

Differential Scanning Calorimetry (DSC) Analysis
The DSC thermograms of the control and Biofield Energy Treated zinc chloride are presented in Figure 5. The decomposition temperature and enthalpy of decomposition of both the control and Biofield Energy Treated zinc chloride are presented in Table 4. A significant increase (15.75%) in the decomposition temperature (T decomp ) was observed in the Biofield Energy Treated zinc chloride (489.06°C) as compared to the control sample (422.50°C). The onset and endset decomposition temperatures of the Biofield Energy Treated sample were significantly increased by 16.61% and 16.05%, respectively compared with the control sample. The control zinc chloride exhibited the enthalpy decomposition of 55.42 J/g, whereas the Biofield Energy Treated sample showed 131.40 J/g. The results as shown in Table 4, indicated a significant increase in the enthalpy of decomposition by 137.10% with respect to the control sample. It is anticipated that Biofield Energy Treatment might enhance the internal energy in zinc chloride that leads to increase in the enthalpy of 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 shelf-life for any pharmaceutical formulations [54].

Conclusions
The current study revealed that the Biofield Energy Healing Treatment (The Trivedi Effect ® ) had the notable effects on the physicochemical and thermal properties of zinc chloride. The PXRD data displayed the significant alteration of the crystallite size and relative intensities of the XRD peaks in The Trivedi Effect ® sample with respect to the control sample. The average crystallite size of The Trivedi Effect ® Treated sample was significantly enhanced by 41.22% with respect to the control sample. The particle size analysis indicated a significant increment in the particle size of the treated zinc chloride at d 10 , d 50 , and d 90 by 8.33%, 6.27%, and 1.83%, respectively with respect to the control sample. Consequently, the surface area analysis Healing Treatment on the Physical, Structural, and Thermal Properties of Zinc Chloride indicated 5.9% reduction in surface area of the treated sample with respect of the control sample. The weight loss of the treated zinc chloride in 1 st and 2 nd step degradation was significantly decreased by 13.98% and 2.43%, respectively compared with the control sample. The DSC analysis exhibited a significant increase in the decomposition temperature by 15.75%, along with a noticeable enhancement of the enthalpy of decomposition by 137.10% in the treated zinc chloride with compared with the control sample. The TGA and DSC analysis exhibited that the treated zinc chloride was found to be thermally more stable compared with the control sample. Briefly, The Trivedi Effect ® -Energy of Consciousness Healing Treatment might introduce a new thermally stable polymorphic form of zinc chloride that can have improved powder flowability, size, and appearance. The treated sample could be more stable during manufacturing, delivery or storage conditions than the untreated sample. Hence, the 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, immunological disorders, aging, stress, cancer, Wilson's disease, viral diseases, parakeratosis, hypogeusia, anorexia, dysosmia, geophagia, hypogonadism, growth retardation, etc.