ANMC Journal

1.       Introduction

6-mercaptopurine is an antimetabolite antineoplastic agent. It has immunosuppressant properties. The mechanism of action involves interference with nucleic acid synthesis by inhibiting purine metabolism [1,2].  It is used as a medication for cancer and autoimmune diseases, i.e., acute lymphocytic leukaemia, chronic myeloid leukaemia, Crohn's disease, and ulcerative colitis [3-5]. It was approved for medical use in the United States since 1953 and also listed as Essential Medicines by the World Health Organization [6]. Common side effects associated with the mercaptopurine therapy are diarrhoea, nausea, vomiting, loss of appetite, mouth sores, stomach and abdominal pain, fatigue, weakness, fever, sore throat, skin rash, pinpoint red spots on the skin, darkening of the skin, yellowing of eyes or skin, hair loss, easy bruising or bleeding, dark urine, black or tarry stools, bloody stools, bloody urine, painful or difficult urination, may suppress the production of blood cells (both white and red blood cells) and can be linked to the genetic polymorphisms [7-9]. Mercaptopurine is used in the pharmaceutical formulations for oral medications in the form of tablets and in liquid suspension [10-12]. Mercaptopurine is insoluble in water, acetone, chloroform, and diethyl ether, but slightly soluble in dilute sulphuric acid; and soluble in hot alcohol and dilute alkali solutions [12]. Many scientific communities throughout the globe are researching ways to improve solubility, dissolution, absorption, and bioavailability of pharmaceutical and nutraceutical compounds because the physicochemical properties of pharmaceutical or nutraceutical compounds are very important [13].

The Trivedi Effect^®-Biofield Energy Healing Treatment has shown a significant impact in the alterations of physicochemical properties such as crystallite size, particle size, surface area, thermal behaviour, and bioavailability profile of pharmaceutical and nutraceutical compounds [14-18]. Every living organism possesses a unique para-dimensional electromagnetic field around the body which generated from the continuous movement of the electrically charged particles (ions, cells, etc.), blood flow, and heart moments inside the body known as “Biofield” (Putative Energy Field). The Trivedi Effect^® (Biofield Energy) is a natural and is the only scientifically proven phenomenon in which a person can harness this inherently intelligent energy from the Universe and transmit it anywhere on the planet through the possible mediation of neutrinos [19]. Due to the increasing beneficial effects of Complementary and Alternative Medicine (CAM) therapies and integrative healthcare approaches including the Biofield Energy Therapies, alternative medicine is an emerging field treatment methods against various health conditions [20,21]. CAM therapies has been recognized by National Center of Complementary and Integrative Health (NCCIH) along with other therapies, which include Ayurvedic medicine, naturopathy, homeopathy, Tai Chi, Qi Gong, acupuncture, acupressure, healing touch, Reiki, hypnotherapy, Rolfing, etc. The CAM has been accepted by the most of the U.S. population because of several advantages [22,23].

Similarly, the Trivedi Effect^®-Consciousness Energy Healing Treatment has significant impact on the transformation in non-living materials and living organisms, and the outcomes were published in numerous peer-reviewed scientific journals. The Trivedi Effect^®-Biofield Energy Treatment has the unprecedented capability to transform the physicochemical, structural, and behavioural properties of organic compounds [24,25], metals [26,27], ceramics [28], polymers [29], microorganisms [30,31], living cells [32,33], to improve the overall productivity of crops [34,35], and alter several qualities of metals and ceramics [36,37]. Alice Branton (the USA) is a renowned Biofield Energy Healer.  She has the ability to harness Life Force from nature and transmit it to living and non-living object(s), anywhere in the world through thought intention, to change their characteristics, transform their behaviour and enhance their function. She is on a mission to winner the breakthrough science of Life Force Energy and its applications in pharmaceutical and nutraceuticals product research to usher in a new era of human health and wellness. The current study was carried out to evaluate the impact of the Trivedi Effect^®-Consciousness Energy Healing Treatment by Alice Branton on the physicochemical, thermal, and behavioural properties of 6-mercaptopurine using powder x-ray diffraction (PXRD), particle size analysis (PSA), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA)/ differential thermogravimetric analysis (DTG).

2.       Materials and Methods

2.1.   Chemicals and Reagents

6-Mercaptopurine monohydrate was purchased from Tokyo Chemical Industry Co., Ltd., Japan. All other chemicals used during the experiments were of analytical grade available in India.

2.2.   Consciousness Energy Healing Treatment Strategies

6-mercaptopurine was the test sample for the experiment, which divided into two equal parts. One part of mercaptopurine was treated with The Trivedi Effect^®-Energy of Consciousness Healing Treatment remotely under standard laboratory conditions for 3 minutes and known as The Trivedi Effect^® Treated (or Biofield Energy Treated) sample. This Biofield Energy Treatment was provided through the healer’s unique energy transmission process by the renowned Biofield Energy Healer, Alice Branton, USA, to the mercaptopurine. However, the second part of mercaptopurine was considered as a control sample (to this no Biofield Energy Treatment was provided). Further, the control sample was treated with a “sham” healer for the comparison purposes. The “sham” healer did not have any knowledge about the Biofield Energy Treatment. After treatment, the Biofield Energy Treated and untreated samples were kept in sealed conditions and characterized using PXRD, PSA, DSC, and TGA techniques.

2.3.   Characterization

2.3.1.         Powder X-ray Diffraction (PXRD) Analysis

The PXRD analysis of mercaptopurine was performed with the help of Rigaku MiniFlex-II Desktop X-ray diffractometer (Japan) [38,39]. The Cu Kα radiation source tube output voltage used was 30 kV, and tube output current was 15 mA. Scans were performed at room temperature. The average size of individual crystallites was calculated from PXRD data using the Scherrer’s formula (1):

                                                                                G = kλ/βcosθ                               (1)

Where k is the equipment constant (0.94), G is the crystallite size in nm, λ is the radiation wavelength (0.154056 nm for Kα1 emission), β is the full-width at half maximum (FWHM), and θ is the Bragg angle [40].

The percent change in crystallite size (G) of mercaptopurine was calculated using the following equation 2:

Where G_Control and G_Treated are the crystallite size of the control and the Biofield Energy Treated samples, respectively.

2.3.2.         Particle Size Analysis (PSA)

The particle size analysis of mercaptopurine was conducted on Malvern Mastersizer 2000, (UK) with a detection range between 0.01 µm to 3000 µm using wet method [41,42]. The sample unit (Hydro MV) was filled with a dispersant medium (sunflower oil) and operated the stirrer at 2500 rpm. The PSA analysis of mercaptopurine was performed to obtain the average particle size distribution. Where d (0.1) μm, d (0.5) μm, d (0.9) μm represent particle diameter corresponding to 10%_,50% and 90% of the cumulative distribution. D(4,3) represents the average mass-volume diameter, and SSA is the specific surface area (m²/g). The calculations were done by using software Mastersizer Ver. 5.54.

The percent change in particle size (d) for mercaptopurine at below 10% level (d₁₀)_, 50% level (d₅₀)_, 90% level (d₉₀), and D (4,3) was calculated using the following equation 3:

Where d_Control and d_Treated are the particle size (μm) for at below 10% level (d₁₀)_, 50% level (d₅₀)_, and 90% level (d₉₀) of the control and the Biofield Energy Treated samples, respectively.

The percent change in surface area (S) was calculated using the following equation 4:

Where S_Control and S_Treated are the surface area of the control and the Biofield Energy Treated mercaptopurine, respectively.

2.3.3.         Differential Scanning Calorimetry (DSC)

The DSC analysis of mercaptopurine was performed with the help of DSC Q200, TA instruments. The sample of ~1-2 mg was loaded to the aluminium sample pan at a heating rate of 10ºC/min from 30°C to 350°C [41,42]. The % change in melting point (T) was calculated using the following equation 5:

Where T_Control and T_Treated are the melting point of the control and treated samples, respectively.

The percent change in the latent heat of fusion (ΔH) was calculated using the following equation 6:

Where ΔH_Control and ΔH_Treated are the latent heat of fusion of the control and treated mercaptopurine, respectively.

2.3.4.         Thermal Gravimetric Analysis (TGA) / Differential Thermogravimetric Analysis (DTG)

TGA/DTG thermograms of mercaptopurine were obtained with the help of TGA Q50 TA instruments. The sample of ~2-5 mg was loaded to the platinum crucible at a heating rate of 10^ºC/min from 25^°C to 1000^°C with the recent literature [41,42]. The % change in weight loss (W) was calculated using the following equation 7:

Where W_Control and W_Treated are the weight loss of the control and the Biofield Energy Treated mercaptopurine, respectively.

The % change in maximum thermal degradation temperature (T_max) (M) was calculated using the following equation 8:

Where M _Control and M _Treated are the T _max values of the control and the Biofield Energy Treated mercaptopurine, respectively.

3.       Results and Discussion

3.1.   Powder X-ray Diffraction (PXRD) Analysis

The PXRD diffractograms of the control and the Biofield Energy Treated sample showed sharp and intense peaks (Figure 1). These sharp and intense peaks in the chromatograms indicated that both the samples were crystalline. The PXRD diffractograms of the control and the Biofield Energy Treated mercaptopurine samples showed the highest peak intensity at 2θ near to 27.47° and 27.36° (Table 1, entry 10). The peak intensities of the Biofield Energy Treated mercaptopurine were significantly altered compared to the control sample. Overall, the peak intensities of the Biofield Energy Treated sample were significantly altered in the range from -63.38% to 3.85% compared to the control sample (Table 1). Likewise, the crystallite sizes of the Biofield Energy Treated sample were significantly altered in the range from -55.26% to 228.26% compared to the control sample (Table 1). Overall, the average crystallite size of the Biofield Energy Treated mercaptopurine (320.65 nm) was significantly increased by 8.16% compared with the control sample (296.45 nm) (Table 1)

The peak intensity of each diffraction face on the crystalline compound changes according to the crystal morphology [43] and alterations in the PXRD pattern provide the proof of polymorphic transitions [44,45]. Therefore, any change observed in the crystallite sizes and peak intensities indicated the modification of the crystal morphology of the Biofield Energy Treated mercaptopurine compared to the control sample. The Trivedi Effect^®-Consciousness Energy Healing Treatment probably produced a new polymorphic form of mercaptopurine through the Biofield Energy via neutrino oscillations [19]. Different polymorphic forms of pharmaceuticals have the significant effects on the drug performance, such as bioavailability, therapeutic efficacy, and toxicity, because of their thermodynamic and physicochemical properties like melting point, energy, stability, and especially solubility, are different from the original form [46,47]. Thus, it can be assumed that the Trivedi Effect^® Treated mercaptopurine would be more efficacious in pharmaceutical formulations.

3.2.   Particle Size Analysis (PSA)

The particle size distribution analysis of both the samples was performed to analyse the change in the particle size, and surface area of the Biofield Energy Treated mercaptopurine compared to the control sample, and the data are presented in (Table 2). The particle size values of the control mercaptopurine at d₁₀, d₅₀, d₉₀, and D (4,3) were 34.82 µm, 99.13 µm, 190.68 µm, and 107.03 µm, respectively. Similarly, the particle sizes of the Biofield Energy Treated mercaptopurine at d₁₀, d₅₀, d₉₀, and D (4,3) were 25.55 µm, 74.91 µm, 157.20 µm, and 84.11 µm, respectively. The particle size values in the Biofield Energy Treated mercaptopurine were significantly decreased at d₁₀, d₅₀, d₉₀, and D (4,3) by 26.63%, 24.43%, 17.56%, and 21.41% compared to the control sample. Therefore, the specific surface area of the Biofield Energy Treated mercaptopurine (0.134 m²/g) was significantly increased by 38.57% compared to the control sample (0.0967 m²/g). Hence, it can be assumed that the Trivedi Effect^®-Consciousness Energy Healing Treatment might act as an external force for the conversion of the larger particles into smaller particles, thus increasing the surface area. It was reported that the particle size, shape, and surface area impact the solubility, dissolution rate, absorption, bioavailability, and even the therapeutic efficacy of a drug [47,48]. The solubility of mercaptopurine is almost insoluble in water, acetone, chloroform, and diethyl ether, whereas slightly soluble in dilute sulphuric acid; soluble in hot alcohol and dilute alkali solutions [12]. Thus, it is anticipated that the Biofield Energy Treated mercaptopurine would show the better therapeutic properties in pharmaceutical formulations by increasing solubility, dissolution, and absorption, which would be better for the pharmaceutical industry when used as a raw material for manufacturing.

3.3.   Differential Scanning Calorimetry (DSC) Analysis

DSC thermograms of both control and the Biofield Energy Treated mercaptopurine are presented in (Figure 2). The DSC thermograms of the control and the Biofield Energy Treated mercaptopurine showed two sharp endothermic peaks in the thermograms (Figure 2). The 1^st peak indicated the peak of evaporation of the bounded water from 6-mercaptopurine monohydrate. Similarly, the 2^nd peak indicated the melting point of 6-mercaptopurine. The thermogram pattern and melting point closely matched to the literature reported data [49]. The evaporation and melting point of the Biofield Energy Treated mercaptopurine were slightly altered by 0.49% and -0.44%, respectively compared with the control sample (Table 3).

The latent heat of evaporation (∆E_vaporation) of the Bio Field Energy Treated mercaptopurine (303.3 J/g) was slightly decreased by 1.46% compared with the control sample (307.8 J/g) (Table 3). However, the latent heat of fusion (∆H_fusion) of the Bio Field Energy Treated mercaptopurine (147.5 J/g) was significantly increased by 49.97% compared with the control sample (98.35 J/g) (Table 3). As per the literature, any change in the latent heat of fusion can be attributed to the disrupted molecular chains and the crystal structure [50]. Thus, it can be predicted that the Trivedi Effect^®-Consciousness Energy Healing Treatment might be responsible for the disruption of the molecular chains and crystal structure of mercaptopurine. The evaporation and melting temperatures of the Biofield Energy Treated samples did not alter much compared to the control sample, but the significant increase in the latent heat of fusion indicated that the thermal stability of the Biofield Energy Treated sample was increased compared to the control sample.

3.4.   Thermal Gravimetric Analysis (TGA)/Differential Thermogravimetric Analysis (DTG)

The TGA thermograms of the control and the Biofield Energy Treated mercaptopurine samples showed three steps of thermal degradation (Figure 3). The total weight loss of the Biofield Energy Treated mercaptopurine was decreased by 11.45% compared to the control sample (Table 4). Therefore, the residue amount was significantly increased by 160.07% in the Biofield Energy Treated mercaptopurine compared to the control sample (Table 4).

The DTG of the control and the Biofield Energy Treated mercaptopurine also showed three peaks in the thermograms (Figure 4). The maximum thermal degradation temperature (T_max) of the 1^st, 2^nd and 3^rd peaks of the Biofield Energy Treated sample altered by -2.26%, -0.46%, and 0.33% compared with the control sample (Table 4). Overall, TGA/DTG analysis of mercaptopurine samples revealed that the thermal stability of the Biofield Energy Treated sample was increased compared with the control sample. The improved thermal stability would improve the quality and self-life of mercaptopurine in the pharmaceutical formulations.

4.       Conclusion

The experimental results confirmed that the Trivedi Effect^®-Consciousness Energy Healing Treatment has significant effects on the particle size, surface area, and thermal properties of 6-mercaptopurine. The PXRD analysis indicated that the peak intensities and crystallite sizes of the Biofield Energy Treated mercaptopurine were significantly altered in the range from -63.38% to 3.85% and -55.26% to 228.26%, respectively compared to the control sample. The average crystallite size of the Biofield Energy Treated mercaptopurine was significantly increased by 8.16% compared with the control sample. The particle size values in the Biofield Energy Treated mercaptopurine were significantly decreased by 26.63%, 24.43%, 17.56%, and 21.41% at d₁₀, d₅₀, d₉₀, and D (4,3) compared to the control sample. Therefore, the specific surface area of the Biofield Energy Treated mercaptopurine was significantly increased by 38.57% compared to the control sample. The significant increase in the ∆H_fusion by 49.97% in the Biofield Energy Treated mercaptopurine indicated that the thermal stability of the Biofield Energy Treated sample was more compared to the control sample. The total weight loss was decreased by 11.45%; however the residue amount was significantly increased by 160.07% in the Biofield Energy Treated sample compared with the control sample. The Trivedi Effect^®-Consciousness Energy Healing Treatment might lead to the generation of a new polymorphic form of mercaptopurine which would offer better solubility, dissolution, absorption, and bioavailability compared with the control sample. The Trivedi Effect^®-Consciousness Energy Healing Treated mercaptopurine would be more efficacious in pharmaceutical formulations that might offer better therapeutic response against acute lymphocytic leukemia, chronic myeloid leukemia, Crohn's disease, and ulcerative colitis, etc.

5.       Acknowledgements

The authors are grateful to Central Leather Research Institute, SIPRA Lab. Ltd., Trivedi Science, Trivedi Global, Inc., Trivedi Testimonials, and Trivedi Master Wellness for their assistance and support during this work.

6.       Conflict of Interest

Authors declare no conflict of interest.

Figure 1: PXRD diffractograms of the control and the Biofield Energy Treated mercaptopurine.

Figure 2: DSC thermograms of the control and the Biofield Energy Treated mercaptopurine.

Figure 3: TGA thermograms of the control and the Biofield Energy Treated mercaptopurine.

Figure 4: DTG thermograms of the control and the Biofield Energy Treated mercaptopurine.

Table 1: PXRD data for the control and the Bio field Energy Treated mercaptopurine.

 Table 2: Particle size distribution of the control and the Biofield Energy Treated mercaptopurine.

Table 3: DSC data for both control and the Bio field Energy Treated samples of mercaptopurine.

Table 4: TGA/DTG data of the control and the Biofield Energy Treated samples of mercaptopurine.

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