# Spectroscopic Characterization of Chloramphenicol and Tetracycline: An Impact of Biofield Treatment

Mahendra Kumar Trivedi, Shrikant Patil, Harish Shettigar, Khemraj Bairwa, Snehasis Jana

► To cite this version:

Mahendra Kumar Trivedi, Shrikant Patil, Harish Shettigar, Khemraj Bairwa, Snehasis Jana. Spectroscopic Characterization of Chloramphenicol and Tetracycline: An Impact of Biofield Treatment. Pharmaceutica Analytica Acta, 2015, 6 (7), pp.395. hal-01376485

HAL Id: hal-01376485

https://hal.science/hal-01376485

Submitted on 5 Oct 2016

HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers.

L'archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d'enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés.

<figure><img src="image_1.png" /></figure>

Distributed under a Creative Commons Attribution 4.0 International License
# Spectroscopic Characterization of Chloramphenicol and Tetracycline: An Impact of Biofield Treatment

Mahendra Kumar Trivedi<fnref n="1" />, Shrikant Patil<fnref n="1" />, Harish Shettigar<fnref n="1" />, Khemraj Bairwa<fnref n="2" /> and Snehasis Jana<fnref n="2" />*

## Abstract

**Objective:** Chloramphenicol and tetracycline are broad-spectrum antibiotics and widely used against variety of microbial infections. Nowadays, several microbes have acquired resistance to chloramphenicol and tetracycline. The present study was aimed to evaluate the impact of biofield treatment for spectroscopic characterization of chloramphenicol and tetracycline using FT-IR and UV-Vis spectroscopy.

**Methods:** The study was performed in two groups (control and treatment) of each antibiotic. The control groups remained as untreated, and biofield treatment was given to treatment groups.

**Results:** FT-IR spectrum of treated chloramphenicol exhibited the decrease in wavenumber of NO₂ from 1521 cm⁻<fnref n="1" /> to 1512 cm⁻<fnref n="1" /> and increase in wavenumber of C=O from 1681 cm⁻<fnref n="1" /> to 1694 cm⁻<fnref n="1" /> in acylamino group. It may be due to increase of conjugation effect in NO₂ group, and increased force constant of C=O bond. As a result, stability of both NO₂ and C=O groups might be increased in treated sample as compared to control. FT-IR spectrum of treated tetracycline showed the downstream shifting of aromatic C-H stretching from 3085-3024 cm⁻<fnref n="1" /> to 3064-3003 cm⁻<fnref n="1" /> and C=C stretching from 1648-1582 cm⁻<fnref n="1" /> to 1622-1569 cm⁻<fnref n="1" /> and up shifting of C-N stretching from 965 cm⁻<fnref n="1" /> to 995 cm⁻<fnref n="1" />. It may be due to enhanced conjugation effect in tetracycline, and increased force constant of C-N (CH₃) bond of tetracycline as compared to control. The results indicated the enhanced stability of treated tetracycline as compared to control. UV-Vis spectra of biofield treated chloramphenicol and tetracycline showed the similar lambda max (λmax) to their respective control. It revealed that the chromophore groups of both antibiotics remained same as control after the biofield treatment.

**Conclusion:** Based on FT-IR spectroscopic data, it is speculated that due to increase in bond strength and conjugation effect after biofield treatment, the chemical stability of both the drugs might be increased as compared to control.

**Keywords:** Chloramphenicol; Tetracycline; Biofield treatment; Fourier transform infrared spectroscopy; Ultraviolet spectroscopy

## Introduction

Chloramphenicol and tetracycline are structurally dissimilar broad-spectrum antibiotics that commonly act by inhibiting the protein synthesis in microbes. These are extensively used in several microbial infections including Gram-negative, Gram-positive bacteria, chlamydiae, and rickettsiae [1-3]. Chloramphenicol reversibly binds to 50S ribosomal subunit of microbes and prevents the binding of aminoacyl tRNA to 50S ribosomal subunit. Thus, it inhibits protein synthesis in bacteria, which is essential for bacterial growth [3,4]. Despite of its broad spectrum antimicrobial activities, it also has some adverse effects such as gray baby syndrome, aplastic anemia, and bone marrow depression [3,5]. Nowadays, chloramphenicol is not much effective due to development of resistance in the variety of microbes. Microbial resistances against chloramphenicol occur by several mechanisms like enzymatic (acetyltransferases and phosphotransferases) inactivation; decreasing the membrane permeability, mutation/modification in target site, and presence of efflux pumps [6]. Tetracycline exerts both bacteriostatic and bactericidal mode of action against the majority of aerobic, anaerobic, Gram-negative, and Gram-positive bacteria. It binds with 30S ribosomal subunit of microbes and block the binding of activated aminoacyl-tRNA to the A site of ribosome. Thus, it blocks the insertion of new amino acids to the nascent peptide chain [2,7,8]. Microbes acquire resistance to tetracycline by evolving the efflux pump and/or by ribosomal protection protein [9,10].

Antimicrobial resistance is now conceiving as global threat; as a result of need of new antimicrobial drugs is constantly increasing.

Therefore, global scientific community has attempted to discover a concept to overcome the microbial resistance i.e. reintroduction of previously used antibiotics active against multi drug resistant (MDR) bacteria. Therefore, several antimicrobial agents like chloramphenicol, tetracycline, etc are reemerging after some modifications as valuable alternatives for the treatment of difficult-to-treat microbial infections [11,12].

Stability of drug is of great importance for its efficacy. In addition, drug degradation may lead to byproducts formation. Abachi et al. showed that stability of chloramphenicol was independent of pH between 4 to 6.2, and it was susceptible to hydrolysis in aqueous media [13]. Lv et al. suggested that chloramphenicol was not stable in suspension form [14]. Liang et al evaluated the stability of tetracycline in methanol solution using UV-Visible spectroscopy, HPLC, and TLC methods. The report showed that tetracycline decomposed quickly

*Corresponding author: Snehasis Jana, Trivedi Science Research Laboratory Pvt. Ltd., Hall-A, Chinar Mega Mall, Chinar Fortune City, Hoshangabad Rd., Bhopal- 462026, Madhya Pradesh, India, Tel: +91-755-6660006; Email: publication@trivedisrl.com

Received June 10, 2015; Accepted July 21, 2015; Published July 24, 2015

Citation: Trivedi MK, Patil S, Shettigar H, Bairwa K, Jana S, et al. (2015) Spectroscopic Characterization of Chloramphenicol and Tetracycline: an Impact of Biofield. Pharm Anal Acta 6: 395. doi:10.4172/21532435.1000395

Copyright: © 2015 Trivedi MK, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

<footnote n="1">¹Trivedi Global Inc., 10624 S Eastern Avenue Suite A-969, Henderson, NV 89052, USA</footnote>

<footnote n="2">²Trivedi Science Research Laboratory Pvt. Ltd., Hall-A, Chinar Mega Mall, Chinar Fortune City, Hoshangabad Rd., Bhopal-462026, Madhya Pradesh, India</footnote>
with the influence of light and atmospheric oxygen, and formed several
degradation products [15,16]. Therefore, an alternative approach is needed
that can increase the shelf life of poorly stable drug.

Recently an alternative approach, biofield treatment is recognized to
change the various physical and structural properties at the atomic level
of various living and non-living things. It is well established that electrical
current coexist along with the magnetic field inside the human body in the
form of vibratory energy particles like ions, protons, and electrons [16,17].
Willem Einthoven discovered an electrocardiography in 1924 to measure
the human biofield. Later on, Harold Saton Burr gave the hypothesis that
every dynamic process in the human body had an electrical significance.
Recently, it confirmed that all the electrical process happening in human
body generates magnetic field [18]. It can be observed using some
medical technologies such as electrocardiography, electromyography,
and electroencephalogram. The electromagnetic field of the human body
is known as biofield and energy linked with this field is called biofield
energy [19-21]. Thus, a human has the ability to harness the energy from
environment or universe and can transmit into any living or nonliving
object around this globe. The object(s) always receive the energy and
responding into useful way; this process is known as biofield treatment
(The Trivedi Effect*). Mr. Trivedi's biofield treatment has altered the
physicochemical and structural properties of metals and ceramics [22-24].
The growth and anatomical characteristics of medicinal plant also changed
after biofield treatment [25]. The biofield treatment enhanced the yield and
quality of agriculture product [26]. Moreover, the changes in antimicrobial
susceptibility and biotype number of pathogenic microbe have been
reported after biofield treatment [27].

Conceiving the concept of antimicrobial reuses, the present study was
aimed to evaluate the impact of biofield treatment on spectral properties of
two antibiotics i.e. chloramphenicol and tetracycline.

Materials and Methods

Study design

The chloramphenicol and tetracycline (Figure 1) samples were
procured from Sigma-Aldrich, MA, USA; and each antibiotic was
divided into two parts: control and treatment. The control samples
remained as untreated, and treatment samples were handed over in
sealed pack to Mr. Trivedi for biofield treatment under laboratory
condition. Mr. Trivedi provided this treatment through his energy
transmission process to the treatment groups without touching the
objects. After that, the control and treated samples of each antibiotic
were analyzed using Fourier transform infrared (FT-IR) spectroscopy
and Ultraviolet-Visible (UV-Vis) spectroscopy.

FT-IR spectroscopic characterization

FT-IR spectra were recorded on Shimadzu's Fourier transform
infrared spectrometer (Japan) with frequency range of 4000-500 cm⁻¹.
The FT-IR spectral analysis of chloramphenicol and tetracycline were
carried out to evaluate the impact of biofield treatment at atomic level
like bond strength, stability, and rigidity of chemical structure [28].

UV-Vis spectroscopic analysis

UV-Vis spectra of chloramphenicol and tetracycline were acquired
on a Shimadzu UV-2400 PC series spectrophotometer with 1 cm quartz
cell and a slit width of 2.0 nm. The analysis was carried out at wavelength
range of 200-400 nm. UV-Vis spectroscopic analysis was performed to
evaluate the effect of biofield treatment on structural property of tested
antibiotics (chloramphenicol and tetracycline) [28].

**Results and Discussion**

FT-IR spectroscopic analysis

FT-IR spectra of control and treated chloramphenicol are shown
in Figure 2. FT-IR spectrum of control sample showed absorption
peaks at 3352-3246 cm⁻¹ that were assigned to O-H and N-H stretching,
respectively. IR peaks at 3081 cm⁻¹ were assigned to aromatic C-H
stretching. Vibrational peaks at 1681 and 1559 cm⁻¹ were assigned to
C=O and C=C stretching, respectively. Further, the NO₂ and C-Cl
stretching were observed at 1521 cm⁻¹ and 662 cm⁻¹, respectively; and
N-H bending was appeared at 1518 cm⁻¹. The FT-IR data of control
chloramphenicol was well supported by the literature data [29].

The FT-IR spectrum of treated sample of chloramphenicol showed
IR absorption peaks for O-H and N-H stretching at 3243 cm⁻¹ and
aromatic C-H stretching was appeared at 3079 cm⁻¹. The vibrational

<figure><img src="image_2.png" /><figcaption>Figure 1: Chemical structure of (a) chloramphenicol and (b) tetracycline.</figcaption></figure>

<figure><img src="image_3.png" /><figcaption>Figure 2: FT-IR spectra of chloramphenicol (control and treated).</figcaption></figure>
peak of carbonyl group (C=O) and C=C stretching were attribute to IR peaks at 1694 and 1560 cm⁻¹, respectively. Additionally, NO₂ stretching and N-H bending peaks were appeared at 1512 and 1510 cm⁻¹, respectively; and C-Cl stretching was appeared at 667 cm⁻¹.

Altogether, the result suggested that C-H and C=C stretching peaks of aromatic ring were observed in similar frequency region in both control and treated sample of chloramphenicol. It indicated that stability and rigidity of phenyl ring was retained in treated sample as like to control. The vibrational peaks for C=O stretching (acylamino group) was observed towards higher frequency region i.e. from 1681 to 1694 cm⁻¹ that may be due to an increase in force constant of C=O group in treated chloramphenicol. Further, FT-IR spectra of treated chloramphenicol showed decrease in wavenumbers of NO₂ i.e. from 1521 to 1512 cm⁻¹. It might be occurred due to increase of conjugation effect between NO₂ group and phenyl ring [30]. Force constant is directly proportional to bond strength [28]. As the force constant of C=O was increased due to biofield treatment, the stability of acylamino moiety in treated chloramphenicol should also be increased. Additionally, the conjugation effect between NO₂ group and phenyl ring may lead to increase in chemical stability of treated chloramphenicol as compared to control.

The FT-IR spectra of control and treated samples of tetracycline are shown in Figure 3. FT-IR spectrum of control sample showed the absorption peaks for N-H and O-H stretching at 3341-3329 cm⁻¹ and aromatic C-H stretching at 3085-3024 cm⁻¹. The vibrational peaks at 2995-2863 cm⁻¹ and 1648-1582 cm⁻¹ were assigned to CH₃ stretching and C=C stretching, respectively. Aromatic C-H bending was appeared at 1458 cm⁻¹ and CH₃ bending was appeared at 1357 cm⁻¹. The aromatic in plane and out plane deformation peaks were appeared at 1247-1000 cm⁻¹ and 567-501 cm⁻¹, respectively. Vibrational peak at 965 cm⁻¹ was assigned to C-N stretching. The FT-IR data of control sample was well supported by reported data [31].

The FT-IR spectra of treated tetracycline showed the vibrational peaks at 3342-3325 cm⁻¹ that were attributed to N-H and O-H stretching. Vibrational peaks at 3064-3003 cm⁻¹ and 2955-2835 cm⁻¹ were assigned to C-H and CH₃ (methyl) stretching, respectively. C=C stretching peaks were appeared at 1622-1569 cm⁻¹. Bending vibrational peaks for C-H and CH₃ groups were appeared at 1454 and 1357 cm⁻¹, respectively. Vibrational peaks at 1247-1000 cm⁻¹, and 995 cm⁻¹ were assigned to C-H in plane deformation and C-N stretching, respectively. The out plane ring deformation peaks were appeared 567-501 cm⁻¹.

The FT-IR data of control and treated tetracycline showed that N-H and O-H stretching peaks are observed in the similar frequency region in both samples. This suggests no changes in the amide and hydroxyl group of treated tetracycline as compared to control. IR absorption peaks for C-H (aromatic) stretching were shifted to lower frequency i.e. from 3085-3024 cm⁻¹ to 3064-3003 cm⁻¹ and likely the CH₃ stretching was shifted to lower frequency i.e. from 2995-2863 cm⁻¹ to 2955-2835 cm⁻¹. IR absorption peak for C=C stretching was also appeared at lower frequency region i.e. from 1648-1582 cm⁻¹ to 1622-1569 cm⁻¹ in treated sample as compared to control. It is well known that resonance or conjugation of C=C double bond or carbonyl group provides the more single bond character to C=C bond that lowers the force constant of respective bond [28,30]. Further, resonance and/or conjugation effect also provide the stability to chemical compound. Therefore, it is speculated that biofield treated tetracycline might be more chemically stable due to enhanced conjugation effect in C and D ring, with respect to control. This could be correlated to enhancement in stability of treated tetracycline with respect of control. The C-N stretching peak in treated

<figure><img src="image_4.png" /><figcaption>**Figure 3:** FT-IR spectra of tetracycline (control and treated).</figcaption></figure>

tetracycline was observed at higher wavenumber than control (i.e. from 965 to 995 cm⁻¹). It could be due to increased bond strength of C-N (CH₃) group that might be increased stability of treated tetracycline as compared to control.

## UV-Vis spectroscopy

UV spectra of control and treated samples of chloramphenicol are shown in Figure 4. It showed no significant change in the lambda max ($λ_{max}$) of treated sample (273.8 nm) as compared to control (272.2 nm), which indicated no changes in chromophore group of treated chloramphenicol with respect to control. UV spectra of control and treated sample of tetracycline are shown in Figure 5. Both spectra showed three absorption peaks at 362.60, 268.80, and 220.60 nm in control sample and 362.60, 268.80, and 221 nm in treated sample, which indicated similar pattern of UV absorbance in both the samples. This suggested no changes in chromophore group of treated tetracycline with respect to control. Overall, the UV spectra of both the antibiotics showed no significant changes in $λ_{max}$ as compared to control. Based on this, it could be concluded functional groups or their position did not altered in treated sample after biofield treatment. To the best of our knowledge, this is the first report showing an impact of biofield treatment on spectral properties (force constant, dipole moment, and bond strength) of chloramphenicol and tetracycline as compared to control.

## Conclusion

Altogether, the FT-IR data showed an alteration in the wavenumber of some functional groups like C=O and NO₂ in chloramphenicol and C=C and C-N (CH₃) in tetracycline with respect to control groups. This could be observed due to some alteration at the atomic level of both
<figure><img src="image_5.png" /><figcaption>**Figure 4:** UV spectra of (a) control and (b) treated chloramphenicol.</figcaption></figure>

<figure><img src="image_6.png" /><figcaption>**Figure 5:** UV spectra of (a) control and (b) treated tetracycline.</figcaption></figure>

antibiotics by the influence of biofield treatment. The results of present study suggest the impact on force constant, bond strength and dipole moment of both antibiotics that may be altered the chemical stability of biofield treatment as compared to control.

## Acknowledgement

The authors would like to acknowledge the whole team of MGV Pharmacy College, Nashik for providing the instrumental facility. Authors would also like to

thank Trivedi science, Trivedi master wellness and Trivedi testimonials for their support during the work.

## References

1. Ruiz NM, Rámirez-Ronda CH (1990) Tetracyclines, macrolides, lincosamides & chloramphenicol. Bol Asoc Med P R 82: 8-17.

2. Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. *Microbiol Mol Biol Rev* 65: 232-260.

3. Feder HM Jr, Osier C, Maderazo EG (1981) Chloramphenicol: A review of its use in clinical practice. *Rev Infect Dis* 3: 479-491.

4. Cundliffe E, McQuillen K (1967) Bacterial protein synthesis: the effects of antibiotics. *J Mol Biol* 30: 137-146.

5. Yunis AA (1988) Chloramphenicol: relation of structure to activity and toxicity. *Annu Rev Pharmacol Toxicol* 28: 83-100.

6. Fernández M, Conde S, de la Torre J, Molina-Santiago C, Ramos JL, et al. (2012) Mechanisms of resistance to chloramphenicol in *Pseudomonas putida* KT2440. *Antimicrob Agents Chemother* 56: 1001-1009.

7. Zakeri B, Wright GD (2008) Chemical biology of tetracycline antibiotics. *Biochem Cell Biol* 86: 124-136.

8. Speer BS, Shoemaker NB, Salyers AA (1992) Bacterial resistance to tetracycline: mechanisms, transfer, and clinical significance. *Clin Microbiol Rev* 5: 387-399.

9. Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. *Microbiol Mol Biol Rev* 65: 232-260.

10. Connell SR, Tracz DM, Nierhaus KH, Taylor DE (2003) Ribosomal protection proteins and their mechanism of tetracycline resistance. *Antimicrob Agents Chemother* 47: 3675-3681.

11. Liaqat I, Sumbal F, Sabri AN (2009) Tetracycline and chloramphenicol efficiency against selected biofilm forming bacteria. *Curr Microbiol* 59: 212-220.

12. Cassir N, Rolain JM, Brouqui P (2014) A new strategy to fight antimicrobial resistance: the revival of old antibiotics. *Front Microbiol* 5: 551.

13. Abachi FT, Bander F, Al-Deeb NN, Nasoh N, Gafar ZM (2010) Formulation and stability studies of chloramphenicol as ophthalmic eye drop. *Tikrit J Pharm Sci* 6: 70-77.

14. Lv FF, Li N, Zheng LQ, Tung CH (2006) Studies on the stability of the chloramphenicol in the microemulsion free of alcohols. *Eur J Pharm Biopharm* 62: 288-294.

15. Liang Y, Denton MB, Bates RB (1998) Stability studies of tetracycline in methanol solution. *J Chromatogr A* 827: 45-55.

16. Planck M (1903) Treatise on Thermodynamics (3rd edition) (english translated by Alexander OGG, Longmans, Green, London, UK).

17. Einstein A (1905) Does the inertia of a body depend upon its energy-content. *Ann Phys* 18: 639-641.

18. Maxwell JC (1865) A dynamical theory of the electromagnetic field. *Phil Trans R Soc Lond* 155: 459-512.

19. Rivera-Ruiz M, Cajavilca C, Varon J (2008) Einthoven's string galvanometer: the first electrocardiograph. *Tex Heart Inst J* 35: 174-178.

20. Burr HS (1957) Bibliography of Harold Saxton Burr. *Yale J Biol Med* 30: 163-167.

21. Rubik B (2002) The biofield hypothesis: its biophysical basis and role in medicine. *J Altern Complement Med* 8: 703-717.

22. Trivedi MK, Patil S, Tallapragada RM (2012) Thought intervention through biofield changing metal powder characteristics experiments on powder characterisation at a PM Plant. *Future Control Automation LNEE* 173: 247-252.

23. Trivedi MK, Patil S, Tallapragada RM (2013) Effect of biofield treatment on the physical and thermal characteristics of silicon, tin and lead powders. *J Material Sci Eng* 2: 125.

24. Trivedi MK, Patil S, Tallapragada RM (2013) Effect of biofield treatment on the physical and thermal characteristics of vanadium pentoxide powders. *J Material Sci Eng S11*: 001.
25. Altekar N, Nayak G (2015) Effect of biofield treatment on plant growth and adaptation. J Environ Health Sci 1: 1-9.

26. Lenssen AW (2013) Biofield and fungicide seed treatment influences on soybean productivity, seed quality and weed community. Agricultural Journal 8: 138-143.

27. Trivedi MK, Patil S (2008) Impact of an external energy on *Staphylococcus epidermis* [ATCC-13518] in relation to antibiotic susceptibility and biochemical reactions-an experimental study. J Accord Integr Med 4: 230-235.

28. Pavia DL, Lampman GM, Kriz GS (2001) Introduction to spectroscopy (3rd edn) Thomson learning, Singapore.

29. Sajan D, Sockalingum GD, Manfait M, Joe IH, Jayakumar VS (2008) NIR-FT Raman, FT-IR and surface-enhanced Raman scattering spectra, with theoretical simulations on chloramphenicol. J Raman Spectros 39: 1772-1783.

30. Seshadri TR, Subba Rao NV, Subrahmanyam B (1968) Effect of conjugation and complex formation on the Raman and I.R. frequencies of the carbonyl group. Proc Indian Acad Sci Sect A 68: 314-323.

31. Gunasekaran S, Varadhan SR, Karunanidhi N (1996) Qualitative analysis on the infrared bands of tetracycline and ampicillin. Proc Indian Natl Sci Acad Part A 62: 309-316.

## Submit your next manuscript and get advantages of OMICS Group submissions

**Unique features:**

* User friendly/feasible website-translation of your paper to 50 world's leading languages

* Audio Version of published paper

* Digital articles to share and explore

**Special features:**

* 400 Open Access Journals

* 30,000 editorial team

* 21 days rapid review process

* Quality and quick editorial, review and publication processing

* Indexing at PubMed (partial), Scopus, EBSCO, Index Copernicus and Google Scholar etc

* Sharing Option: Social Networking Enabled

* Authors, Reviewers and Editors rewarded with online Scientific Credits

* Better discount for your subsequent articles

Submit your manuscript at: http://www.editorialmanager.com/virology