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American Journal of Physical Chemistry 2015 46: 65-70 Published online December 22 2015 http://www.sciencepublishinggroup.com/j/ajpc doi: 10.11648/j.ajpc.20150406.14 ISSN: 2327-2430 Print ISSN: 2327-2449 Online Mass Spectrometry Analysis of Isotopic Abundance of 13 C 2 H or 15 N in Biofield Energy Treated Aminopyridine Derivatives Mahendra Kumar Trivedi 1 Alice Branton 1 Dahryn Trivedi 1 Gopal Nayak 1 Gunin Saikia 2 Snehasis Jana 2 1 Trivedi Global Inc. Henderson USA 2 Trivedi Science Research Laboratory Pvt. Ltd. Bhopal Madhya Pradesh India Email address: publicationtrivedisrl.com S. Jana To cite this article: Mahendra Kumar Trivedi Alice Branton Dahryn Trivedi Gopal Nayak Gunin Saikia Snehasis Jana. Mass Spectrometry Analysis of Isotopic Abundance of 13 C 2 H or 15 N in Biofield Energy Treated Aminopyridine Derivatives. American Journal of Physical Chemistry. Vol. 4 No. 6 2015 pp. 65-70. doi: 10.11648/j.ajpc.20150406.14 Abstract: 2-Aminopyridine 2-AP and 26-diaminopyridine 26-DAP are two derivatives of aminopyridines that act as an important organic intermediates mostly used in medicines dyes and organic sensors. The aim of the study was to evaluate the impact of biofield energy treatment on isotopic abundance ratios of 2 H/ 1 H 13 C/ 12 C or 15 N/ 14 N in aminopyridine derivatives using gas chromatography-mass spectrometry GC-MS. The 2-AP and 26-DAP samples were divided into two parts: control and treated. The control sample remained as untreated while the treated sample was further divided into four groups as T1 T2 T3 and T4. The treated group was subjected to Mr. Trivedi’s biofield energy treatment. The GC-MS spectra of 2-AP and 26- DAP showed five and six m/z peaks respectively due to the molecular ion peak and fragmented peaks of aminopyridine derivatives. The isotopic abundance ratio of 2 H/ 1 H 13 C/ 12 C or 15 N/ 14 N were calculated for both the derivatives and significant alteration was found in the treated samples as compared to the respective control. The isotopic abundance ratio of 2 H/ 1 H 13 C/ 12 C or 15 N/ 14 N in treated samples of 2-AP was decreased by 55.83 in T1 and significantly increased by 202.26 in T4. However in case of 26-DAP the isotopic abundance ratio of 2 H/ 1 H 13 C/ 12 C and 15 N/ 14 N in the treated sample showed a significant increase up to 370.54 in T3 with respect to the control. GC-MS data suggested that the biofield energy treatment on aminopyridine derivatives had significantly altered the isotopic abundance of 2 H 13 C or 15 N in the treated 2-AP and 26- DAP as compared to the control. Keywords: Biofield Energy Treatment 2-Aminopyridine 26-Diaminopyridine Gas Chromatography-Mass Spectrometry 1. Introduction Most of the elements occurred in nature as a mixture of isotopes. The relative abundances of isotopes is different at different places on the earth and remained constant for years. The distribution of contaminant sources of any molecule on a native or global scale can be understood by determining the isotopic abundance ratio 1. Any kinetic process that leads to the local depletion or enhancement of isotopes in organic molecules can be successfully determined using gas chromatography-mass spectrometry GC-MS 2. These deviations from perfect chemical equivalence are termed as isotope effects. The isotopic abundance ratio is commonly reported in terms of atom percent and determined by high resolution mass spectrometry HRMS spectrometry 3. For example 13 C Atom percent 13 C 13 C/ 12 C+ 13 C×100 1 Moreover the rate of chemical reaction may vary with the mass of the nucleus with different isotopic substitutions which slightly affect the partitioning of energy within the molecules 4. The aminopyridines selected for this study are heterocyclic pharmacophores for many bioactive small organic molecules 5. The growing interest of these molecules pharmacologically due to its rare tendency to be oxidized like aniline derivatives 6. Aminopyridines avoid such problems due to its reduced oxidation potential thus make it a safer

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American Journal of Physical Chemistry 2015 46: 65-70 66 alternatives for drug design. Additionally they are advantageous because they have exhibited strong absorption and emission spectra and thus useful as fluorescent tags while investigating the activities on a target 7. Aminopyridines are used as a drug for symptomatic treatment of multiple sclerosis by blocking potassium channels and prolonging action potentials 8 9. It has been used as intermediate for the synthesis of pharmaceutical agents such as piroxicam sulfapyridine tenoxicam and tripelennamine 6 10. Besides diaminopyridines are mostly used for synthesis of dyes cosmetics drugs and explosives 11 12. It is also used as intermediate for the synthesis of epoxy curing agents polyamides and preparation of analgesic phenazo- pyridine hydrochloride 13. Additionally both the aminopyridine derivatives were successfully utilized as organic fluorescence sensors for the detection of metal cation 14. The isotopic abundances of ratios of 2 H/ 1 H 13 C/ 12 C or 15 N/ 14 N could be locally altered by kinetically driven chemical reactions. There is an alternative and well-known approach Mr. Trivedi’s biofield energy treatment also known as The Trivedi Effect ® that can be applied on aminopyridine derivatives to undergo the isotopic changes. The physicochemical properties of various molecules and crystals were already altered by utilizing Mr. Trivedi’s biofield energy treatment 15-17. The National Center for Complementary and Alternative Medicine NCCAM has recommended the use of energy therapy as a part of Complementary and alternative medical therapies CAM in the healthcare sector 18. CAM includes numerous energy- healing therapies in which the biofield therapy is a form of putative energy therapy that is being widely used worldwide to improve the overall health of human beings. Humans have the ability to harness energy from the environment/universe that can be transmitted to any objects around. Based on the previous results achieved by the biofield energy treatments by Mr. Trivedi in various fields biofield energy treated 2-AP and 26-DAP were taken for mass spectroscopy studies to evaluate the isotopic abundance ratio of 2 H/ 1 H 13 C/ 12 C or 15 N/ 14 N PM+1/PM where PM is the primary molecule and PM+1 is isotopic molecule. 2. Experimental 2.1. Materials Both 2-aminopyridine 2-AP and 26-diaminopyridine 26-DAP were procured from S. D. Fine Chemicals Pvt. Limited India. 2.2. Method Each of the 2-AP and 26-DAP sample was distributed into two parts where one part was referred as control and the other part was considered as treated sample. The treated sample was further divided into four groups i.e. T1 T2 T3 and T4 and handed over to Mr. Trivedi for biofield energy treatment under standard laboratory conditions. Mr. Trivedi provided the treatment through his energy transmission process to the treated groups without touching the sample. The control and treated samples were characterized using gas chromatography-mass spectrometry GC-MS. 2.3. GC-MS Spectroscopy The gas chromatography-mass spectroscopy GC-MS analysis was performed on Perkin Elmer/auto system XL with Turbo mass USA having detection limit up to 1 picogram. The GC-MS spectra were obtained in the form of abundance vs. mass to charge ratio m/z. The isotopic abundance ratio PM+1/PM was expressed by its deviation in treated samples as compared to the control. The percentage change in isotopic ratio PM+1/PM was calculated from the following formula: Percent change in isotopic abundance ratio 100 2 where R Treated and R Control are the ratios of intensity at PM+1 to PM in mass spectra of treated and control samples respectively. 3. Results and Discussion 3.1. GC-MS Study of 2-AP Figure 1. GC-MS spectrum of control 2-aminopyridine. The GC-MS spectra of control and treated T1 T2 T3 and T4 samples are presented in Fig. 1 and 2 respectively.

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67 Mahendra Kumar Trivedi et al.: Mass Spectrometry Analysis of Isotopic Abundance of 13C 2H or 15Nin Biofield Energy Treated Aminopyridine Derivatives Mass spectrum of 2-AP is well matched with the reported literature 19. MS spectra showed five peaks including molecular ion peak for both the control and treated samples of 2-AP. Five major peaks at m/z94 67 51 41 and 28 were observed that corresponded to the following ions respectively: C 5 H 6 N 2 + C 4 H 5 N + C 4 H 3 + C 3 H 5 + and C 2 H 4 + ions. The molecular ion peak is the base peak in both control and treated samples at m/z94. The other four peaks were observed in both control and all treated 2-AP samples due to the fragmentation of molecular ion inside mass spectrum. Further m/z67 51 41 and 28 peaks were observed might be due to first reduction of the molecular ion to pyrrole which again fragmented to give buten-3-yne propene and ethane respectively. The intensity ratio and calculated percentage isotopic abundance ratio of all three elements presented in Table 1. Moreover the isotopic abundance ratio PM+1/PM in control and treated 2-AP was plotted in Fig. 3. It was clearly seen from the Table 1 and Fig. 3 that the isotopic abundance ratio of PM+1/PM of 2-AP sample was decreased in T1 55.83 and T2 25.12 but increased significantly in T3 117.19 and T4 202.26 after biofield energy treatment as compared to the control. The decrease in the isotopic abundance ratio of PM+1/PM in T1 and T2 may have nominal effect on the bond energies and reactivity of the molecules of treated samples. However the increased isotopic abundance ratio of PM+1/PM in T3 and T4 may increase the number of higher isotopes PM+1 in the molecule. It may directly affect the bond strength of the C-H N-H and N-C bonds. The increased isotopic abundance ratio of PM+1/PM in the treated T3 and T4 sample may increase µ effective mass and binding energy in the 2-AP molecule with heavier isotopes and this may result in enhance binding energy and stability of the molecule 20. Table 1. GC-MS isotopic abundance analysis result of 2-aminopyridine. Peak Intensity Control Treated T1 T2 T3 T4 m/z of PM 100 100 100 100 100 m/z of PM+1 14.13 6.24 10.58 30.69 42.71 PM+1/ PM 0.141 0.062 0.105 0.306 0.427 Percent change -55.83 -25.12 117.19 202.26 Figure 2. GC-MS spectra of treated 2-aminopyridine samples T1 T2 T3 and T4.

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American Journal of Physical Chemistry 2015 46: 65-70 68 Figure 3. Percent change in the isotopic abundance PM+1/PM of 2- aminopyridine after biofield energy treatment as compared to the control. 3.2. GC-MS Study of 26-DAP Figure 4. GC-MS spectrum of control sample of 26-diaminopyridine. Mass spectra of both control and treated samples were shown in Fig. 4 and 5 respectively. The molecular ion peak was observed at m/z109 in mass spectra of both control and treated samples. The calculated relative intensity ratio of PM+1/PM and percentage abundance ratios are given in Table 2. Total five major peaks were observed in the mass spectra of both control and treated spectra of 26-DAP at m/z109 82 66 55 39 and 28 corresponded to the following ions respectively: C 5 H 7 N 3 + PM C 5 H 8 N + 12- dihydropyridine C 4 H 4 N + pyrrole C 4 H 7 + but-1-ene C 3 H 3 + but-1-ene and C 2 H 4 + ethene ions. In both the molecules 2- AP and 26-DAP molecular ion peak was observed as a base peak. However they exhibited different fragmentation pattern due to the structural and reactivity differences in them. The isotopic abundance ratio of PM+1/PM of control and treated 26-DAP was calculated and presented in Fig. 6. The isotopic abundance ratio of PM+1/PM of treated 26- DAP was increased upto 370.54 in T3 T17.30 T240.24 and T412.82 as compared to the control. The mass spectrum of control 26-DAP is well supported with the reported literature 21. If the lighter isotopes were substituted by heavier isotopes then the effective mass µ of the particular bond is increased and subsequently binding energy will be increased 20. The reverse may also happen if the heavier isotopes part of the chemical bonding were substituted by lighter isotopes in a molecule. Thus the increased isotopic abundance ratio of PM+1/PM in 2-AP and 26-DAP might increase the effective mass and binding energy after biofield energy treatment that may enhance the chemical stability of aminopyridine derivatives. On the contrary the slight decrease in isotopic abundance ratio of PM+1/PM in treated T1 and T2 in 2-AP might reduce the effective mass of the particular bond and binding energy will be decreased. Further the increased PM+1/PM will increase the effective mass which may decrease the reactivity of the aminopyridine derivatives. Table 2. GC-MS isotopic abundance analysis result of 26-diaminopyridine. Peak Intensity Control Treated T1 T2 T3 T4 m/z of PM 100 100 100 99.8 100 m/z of PM+1 6.71 7.2 9.41 31.51 7.57 PM+1/PM 0.067 0.072 0.094 0.315 0.075 Percent change 7.30 40.24 370.54 12.82 Table 3. Possible isotopic bonds in 2-aminopyridine and 26- diaminopyridine. Isotopes Bond Isotope type Reduced mass m Am B /m A + m B 12 C- 12 C Lighter 6.000 13 C- 12 C Heavier 6.260 1 H- 12 C Lighter 0.923 1 H- 13 C Heavier 0.929 2 H- 12 C Heavier 1.710 1 H- 15 N Heavier 0.940 2 H- 14 N Heavier 1.750 15 N- 12 C Heavier 7.200 14 N- 13 C heavier 6.500 15 N- 13 C Heavier 7.170 Where mA- mass of atom A mB- mass of atom B here A may be C or H and so on . Effective mass of some probable isotopic bonds were calculated and presented in Table 3. The result showed that µ of normal 12 C- 12 C µ6 and 1 H- 12 C µ0.923 bonds were increased in case of heavier isotopes i.e. 13 C- 12 C6.26 and 2 H- 12 C1.71 respectively. After biofield treatment bond strength stability and binding energy of the aromatic ring of 2-AP and 26-DAP molecules might be increased due to the higher effective mass µ after biofield energy treatment 22. The decreased reactivity of the aminopyridine derivatives may increase the stability of the aminopyridine based products in pharmaceutical industry by reducing the degradation kinetics in the finished products after production.

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69 Mahendra Kumar Trivedi et al.: Mass Spectrometry Analysis of Isotopic Abundance of 13C 2H or 15Nin Biofield Energy Treated Aminopyridine Derivatives Furthermore the organic molecules used for fluorescence sensors are exposed to UV radiation continuously for a longer period of time. Photo stability is a great concern in order to use them successfully as sensors without degradation. After biofield treatment the stability may enhance due to the presence of higher isotopes an enable them to expose the materials under UV radiation for longer period 14. Figure 5. GC-MS spectra of treated samples of 26-diaminopyridine T1 T2 T3 and T4. Figure 6. Percent change in the isotopic abundance of PM+1/PM of 26- diaminopyridine after biofield energy treatment as compared to the control. 4. Conclusions In summary aminopyridine derivatives 2-AP and 26- DAP were studied with GC-MS under the influence of biofield energy treatment and observed a significant change in isotope abundance of 2 H/ 1 H 13 C/ 12 C or 15 N/ 14 N as compared to the respective control samples. The percent change in isotope abundance ratio of PM+1/PM was increased upto 202.26 in 2-AP T4 while the isotopic abundance ratio was increased significantly upto 370.54 in treated 26-DAP T3 sample. The changes in isotopic abundance ratios have significant impact on bond energies and chemical reactivity of the molecules. Due to the enhancement in the isotopic abundance ratio of PM+1/PM in 2-AP and 26-DAP the reactivity may be reduced significantly by increase in the effective mass m and the binding energy of the treated sample. It can be concluded from the above observations that the enhancement of heavier isotopes in the aromatic ring as well as the functional groups may decrease the reactivity of the aromatic ring and the functional groups of aminopyridine derivatives consequently enabling their utility as pharmacophore in the pharmaceutical industry and as an active material in fluorescence sensors.

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American Journal of Physical Chemistry 2015 46: 65-70 70 Acknowledgements The authors would like to acknowledge the Sophisticated Analytical Instrument Facility SAIF Nagpur for providing the instrumental facility. We are very grateful for the support from Trivedi Science Trivedi Master Wellness and Trivedi Testimonials in this research work. References 1 Muccio Z Jackson GP 2009 Isotope ratio mass spectrometry. Analyst 134: 213-222. 2 Rieley G 1994 Derivatization of organic-compounds prior to gas-chromatographic combustion-isotope ratio mass- spectrometric analysis: Identification of isotope fractionation processes. Analyst 119: 915-919. 3 Weisel CP Park S Pyo H Mohan K Witz G 2003 Use of stable isotopically labeled benzene to evaluate environmental exposures. J Expo Anal Environ Epidemiol 13: 393-402. 4 Hoefs J 2009 Stable Isotope Geochemistry Isotope Fractionation Processes of Selected Elements. Springer-Verlag Berlin Heidelberg. 5 May BCH Zorn JA Witkop J Sherrill J Wallace AC et. al 2007 Structure-activity relationship study of prion inhibition by 2-aminopyridine-3 5-dicarbonitrile-based compounds: Parallel synthesis bioactivity and in vitro pharmacokinetics. J Med Chem 50:65-73. 6 Coleman MD 2010 Human Drug Metabolism. Role of Metabolism in Drug Toxicity. John Wiley Sons Ltd. New York. 7 Araki Y Andoh A Fujiyama Y Hata K Makino J 2001 Application of 2-aminopyridine fluorescence labeling in the analysis of in vivo and in vitro metabolism of dextran sulfate sodium by size-exclusion high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl 753: 209- 215. 8 Korenke AR Rivey MP Allington DR 2008 Sustained- release fampridine for symptomatic treatment of multiple sclerosis. Ann Pharmacother 42: 1458-1465. 9 New Drugs: Fampridine. 2011 Australian Prescriber 34: 119-123. 10 Gonzalez Cabrera D Douelle F Younis Y Feng TS Le Manach C et al. 2012 Structure activity relationship studies of orally active antimalarial 3 5- substituted 2- aminopyridines. J Med Chem 55: 11022-11030. 11 Naixing W Boren C Yuxiang O 1993 Synthesis of N-246- trinitrophenyl-N’- 2 4 dinitrobenzofuroxano-3 5-dinitro- G- diaminopyridine. J Energ Mater 11: 47-50. 12 Schwalbe CH Williams GJB Koetzle TF 1987 A neutron diffraction study of 2 6-Diaminopyridine at 20K. Acta Cryst C 43: 2191-2195. 13 Freeman HG Gillern MF Smith HA 1976 Rapid curing hydrophilic resin compositions. US 3947425 A. 14 Dickson SJ Paterson MJ Willans CE Anderson KM Steed JW 2008 Anion binding and luminescent sensing using cationic rutheniumII aminopyridine complexes. Chem Eur J 14: 7296-7305. 15 Trivedi MK Patil S Tallapragada RM 2012 Thought intervention through bio field changing metal powder characteristics experiments on powder characteristics at a PM plant. Future Control and Automation LNEE 173: 247-252. 16 Trivedi MK Patil S Shettigar H Gangwar M Jana S 2015 Antimicrobial sensitivity pattern of Pseudomonas fluorescens after biofield treatment. J Infect Dis Ther 3: 222. 17 Patil S Nayak GB Barve SS Tembe RP Khan RR 2012 Impact of biofield treatment on growth and anatomical characteristics of Pogostemon cablin Benth.. Biotechnology 11: 154-162. 18 Barnes PM Powell-Griner E McFann K Nahin RL 2004 Complementary and alternative medicine use among adults: United States 2002. Adv Data 343: 1-19. 19 http://webbook.nist.gov/cgi/cbook.cgiIDC504290Mask2 00Mass-Spec. 20 Smith BC 2011 Fundamentals of Fourier transform infrared spectroscopy CRC Press Taylor and Francis Group Boka Raton New York. 21 http://webbook.nist.gov/cgi/cbook.cgiIDC141866Mask2 00Mass-Spec 22 Mook W Vries J 2003-2004 Environmental isotopes in the hydrological cycle principles and applications. International atomic energy agency Vienna 1-271.