Journal of Plant Sciences
2015; 3(6): 285-293
Published online November , 2015 (http://www.sciencepublishinggroup.com/j/jps)
ISSN: 2331-0723 (Print); ISSN: 2331-0731 (Online)
Physicochemical Characterization of Biofield Treated
Orchid Maintenance/Replate Medium
Mahendra Kumar Trivedi
, Alice Branton
, Dahryn Trivedi
, Gopal Nayak
, Ragini Singh
Trivedi Global Inc., Henderson, NV, USA
Trivedi Science Research Laboratory Pvt. Ltd., Bhopal, Madhya Pradesh, India
email@example.com (S. Jana)
To cite this article:
Mahendra Kumar Trivedi, Alice Branton, Dahryn Trivedi, Gopal Nayak, Ragini Singh, Snehasis Jana. Physicochemical Characterization of
Biofield Treated Orchid Maintenance/Replate Medium. Journal of Plant Sciences. Vol. 3, No. 6, 2015, pp. 285-293.
Orchids are used worldwide for indoor decoration, vanilla production, and beverage preparation. They are also
reported for their therapeutic efficacy in brain-related problems. The in vitro micropropagation technique was used for their
propagation using the orchid maintenance/replate (OMR) medium. The current study was based on analysing the effect of
biofield energy treatment on the physicochemical properties of OMR medium. A part of the sample was treated with Mr.
Trivedi’s biofield energy; various physicochemical properties were analyzed and compared with the untreated (control) part.
The X-ray diffraction analysis revealed the decrease in crystallite size of treated sample (132.80 nm) as compared to the
control (147.55 nm). The particle size analysis revealed 20.78% increase in average particle size and 39.29% increase in d
(size below which 99% particles are present) of the treated OMR medium as compared to the control. Moreover, the surface
area of the treated sample was reduced by 3.9%, supporting the data of particle size analysis. The thermal analysis studies
revealed an increase in the thermal stability of the treated OMR medium as compared to the control. The analysis was done by
using differential scanning calorimetry that showed increase in melting point (1.23%) and latent heat of fusion (135.7%); and
thermogravimetric analysis that reported increase in onset temperature and maximum thermal degradation temperature of the
treated sample as compared to the control. Besides, the CHNSO analysis revealed the increase in percentage of nitrogen
(22.22%) as well as the presence of sulphur in the treated sample. The Fourier transform infrared and UV-visible spectroscopy
also showed the differences in the spectra of the treated sample as compared to the control OMR medium. Hence, the overall
data revealed the impact of biofield energy treatment on the physicochemical properties of the treated sample that might be
used in better way in the in vitro culture techniques as compared to the control sample.
Orchid Maintenance/Replate Medium, Biofield Energy Treatment, In vitro Micropropagation,
Complementary and Alternative Medicines
Orchids belonging to family Orchidaceae are widely
used due to their medicinal properties in several countries
around the world . Several kind of research had shown
their therapeutic efficacy in case of hysteria, nervous
irritability, and other brain related dysfunctions. Besides,
several species are reported as a febrifuge, clearing
tapeworms, treating skin diseases, etc. [2, 3]. It was
reported that the biological activity was due to the presence
of alkaloids such as strychnine, morphine, nicotine,
reserpine, etc. . Moreover, the orchid family is also
important for its horticulture uses. It is used for commercial
production of vanilla . In Turkey, it is used in the
preparation of a traditional beverage called as salep . It
has great importance as cut flowers and indoor decoration.
The general method of propagation of orchid is asexual but
it can produce only 2-4 plants per year as it is very slow
process [6, 7]. Hence, in vitro micropropagation technique
is frequently used that produces plantlets using tissue
culture techniques . Micropropagation is the process
used to replicate the plant using plant seed or tissue in the
laboratory under the sterile conditions . The most
important factor in successful tissue culture of plant cells is
286 Mahendra Kumar Trivedi et al.: Physicochemical Characterization of Biofield
Treated Orchid Maintenance/Replate Medium
the composition of the medium. It provides the essential
nutrients for the survival of plant cells or tissues and the
optimum conditions such as pH, osmotic pressure, etc. .
Orchid maintenance/replate (OMR) medium is such type of
medium that consists of accurately defined organic and
inorganic chemical additives in the form of macro- and
micronutrients . The constituents of the medium are
shown in Table 1. It differs from original orchid replate
medium as it contains banana powder to promote rooting
and growth, and agar as a gelling agent that provides
physical support . Despite several advantages, it suffers
the problem of hygroscopicity due to which it needs to be
protected from atmospheric moisture and proper storage
conditions (2-8°C) . Hence, some alternative is needed
that can improve its properties thereby its use as orchid
Table 1. Components of the orchid maintenance/replate medium.
S. No. Ingredient milligram/litre
1 Potassium nitrate 950.00
2 Ammonium nitrate 825.00
3 Calcium chloride.2H
4 Magnesium sulphate 90.34
5 Potassium phosphate monobasic 85.00
6 Manganese sulphate.H
7 Boric acid 3.10
8 Potassium iodide 0.42
9 Molybdic acid (sodium salt).2H
10 Zinc sulphate.7H
11 Copper sulphate.5H
12 Cobalt chloride.6H
13 Ferrous sulphate.7H
14 EDTA disodium salt.2H
15 myo - Inositol 100.00
16 Thiamine hydrochloride 10.00
17 Pyridoxine hydrochloride 1.00
18 Nicotinic acid (Free acid) 1.00
19 Peptone 2000.00
20 Banana powder 30000.00
21 Sucrose 20000.00
22 MES 1000.00
23 Agar 7000.00
24 Activated charcoal 2000.00
MES: 2-(n-morpholino)ethanesulfonic acid; EDTA: Ethylenediamine
The biofield energy treatment is reported to affect the
health of human beings via interacting with their biofield
. The National Centre for Complementary and
Alternative Medicine (NCCAM), which is the part of
National Institute of Health (NIH) also include the energy
medicines as complementary and alternative medicines
(CAM) . It is a putative form of energy that is produced
by own emissions of the body and surrounds the body of all
living organisms. However, the frequency of this energy
depends on the physiological and mental state of the person.
The living systems are continuously exchanging this energy
with their surroundings to maintain themselves [16, 17]. The
non-living things also possess the biofield energy as
everything in the universe made up of same constituents;
however, they are not able to change their energy aura by
more than 2% . Thus, the human has the ability to
harness the energy from the environment and can transmit it
to any living or non-living object. Mr. Trivedi is also known
to possess unique biofield energy, and the treatment is called
as The Trivedi Effect
. It is known for its impact on various
living organisms and non-living materials including
microorganisms , pharmaceutical compounds , and
yield, growth, and anatomical characteristics of plants [21,
22]. Hence, based on the importance of OMR medium and
the outcomes of the biofield energy treatment, the study was
designed to analyse the impact of Mr. Trivedi’s biofield
energy treatment on various physicochemical properties of
the OMR medium.
2. Materials and Methods
Orchid maintenance/replate (OMR) Medium was procured
from HiMedia Laboratories, India. The sample was divided
into two parts and coded as control and treated. Mr. Trivedi’s
biofield energy treatment was provided to the treated part
while no treatment was given to the control part. For
treatment, the treated part was handed over to Mr. Trivedi in
sealed pack under standard laboratory conditions. Mr. Trivedi
provided the treatment to the treated part through his unique
energy transmission process, without touching the sample.
The control and biofield treated samples were further
characterised using various analytical techniques.
2.1. X-ray Diffraction (XRD) Study
The Phillips Holland PW 1710 X-ray diffractometer
system was used to obtain the X-ray powder diffractogram of
control and treated samples. The X-ray generator was
equipped with a copper anode with nickel filter operating at
35kV and 20mA. The XRD system used 1.54056Å
wavelength of radiation. The data were collected from the 2θ
range of 10°-99.99° and a counting time of 0.5 seconds per
step along with a step size of 0.02°.
The crystallite size (G) was calculated from the Scherrer
G = kλ/(bCosθ)
Where, k is constant (0.94), λ is the X-ray wavelength
(0.154 nm), b in radians is the full-width at half of the peak
and θ is the corresponding Bragg’s angle.
Moreover, the percent change in crystallite size was
calculated using the following equation:
Percent change in crystallite size = [(G
denotes the crystallite size of control and
treated powder samples, respectively.
2.2. Particle Size Analysis
For particle size analysis, laser particle size analyzer
SYMPATEC HELOS-BF was used, having a detection
range of 0⋅1-875 µm. Two parameters of particle sizes viz.
(size below which 50% and 99% particles are
Journal of Plant Sciences 2015; 3(6): 285-293 287
present, respectively) were calculated. The percent change
in average particle size (d
) was calculated using following
represents the average
particle size of control and treated samples, respectively.
Similarly, the percent change in particle size d
2.3. Surface Area Analysis
The surface area was measured by the Brunauer–Emmett–
Teller (BET) surface area analyser, Smart SORB 90. The
percent change in surface area was calculated using
%changeinsurfacearea = S
are the surface area of control
and treated samples respectively.
2.4. Thermal Analysis
The thermal stability profile of OMR medium was
analyzed using DSC and TGA/DTG studies. The impact of
biofield treatment was analyzed by comparing the results of
treated sample with that of the control sample.
2.4.1. Differential Scanning Calorimetry (DSC) Study
The DSC analysis of control and treated samples was
carried out using Perkin Elmer/Pyris-1. The samples were
heated at a rate of 10°C/min under air atmosphere (5
mL/min). The thermograms were collected over the
temperature range of 50°C to 300°C.
2.4.2. Thermogravimetric Analysis/Derivative
The effect of temperature on the stability of the control and
treated sample of OMR medium was analyzed using Mettler
Toledo simultaneous thermogravimetric analyser
(TGA/DTG). The heating temperature was selected from
room temperature to 350ºC with a heating rate of 5ºC/min
under air atmosphere.
2.5. CHNSO Analysis
The control and treated samples of OMR medium were
analyzed using CHNSO analyzer using Model Flash EA 1112
series, Thermo Finnigan Italy.
2.6. Fourier Transform-Infrared (FT-IR) Spectroscopic
For FT-IR characterization, the samples were crushed,
mixed with spectroscopic grade KBr and pressed into pellets
with a hydraulic press. The FT-IR spectra were recorded on
Shimadzu’s Fourier transform infrared spectrometer (Japan)
in the frequency range 4000-350 cm
. The FT-IR spectral
analysis was used to determine the effect of biofield energy
on the strength of bonds and stability of compounds present
in OMR medium.
2.7. UV-Vis Spectroscopic Analysis
The UV-Vis spectral analysis was measured using
Shimadzu UV-2400 PC series spectrophotometer. The
spectrum was recorded using 1 cm quartz cell that has a slit
width of 2.0 nm.
3. Results and Discussion
3.1. X-Ray Diffraction (XRD)
The X-ray powder diffractograms of control and treated
samples of OMR medium showed a series of sharp peaks in
the regions of 10º<2θ>40º. In the control sample, the peaks
were observed at 2θ equal to 11.67°, 16.72°, 18.72°, 18.88°,
19.64°, 21.49°, and 25.23°. However, the treated sample
showed the peaks at 2θ equal to 11.61°, 13.05°, 16.67°,
17.73°, 18.74°, 22.04°, 24.64°, and 25.17°. In addition, the
most intense peak in control sample was observed at 2θ equal
to 25.23°; however, in treated sample it was observed at
22.04°. It indicated that the relative intensities of XRD peaks
were altered in the treated OMR medium as compared to the
control sample. Besides, the crystallite size of the control
sample was found as 147.55 nm whereas; in the treated
sample it was found as 132.80 nm. It suggested that
crystallite size of the treated sample was significantly
decreased by 10% as compared to the control. The changes in
the relative intensities of peaks revealed the presence of
microstrain may be due to the biofield treatment. It may
result in change in dislocation densities and atomic
displacements that might be the reason for decreased
crystallite size [23, 24].
3.2. Particle Size Analysis
The particle size of control and treated samples of OMR
medium are presented in Fig. 1. It showed that the d
were 26.03 and 236.92 µm, respectively in the control
sample. However, in treated sample, the d
found as 31.44 and 330.01 µm, respectively. It revealed that
was increased by 20.78% and d
was increased by
39.29% in the treated sample as compared to the control. The
temperature has a significant effect on the particle size of the
sample . Hence, it is presumed that the biofield treatment
may provide some energy to the sample that resulted in
increased particle size as compared to the control sample.
Moreover, the particle size was directly related to the
viscosity and gelling property of the compound . Hence,
the treated OMR medium sample with increased particle size
might improve the gelling property of media, as the large
particles has less tendency to broken down and has stronger
water holding capacity as compared with the small particles.
288 Mahendra Kumar Trivedi et al.: Physicochemical Characterization of Biofield
Treated Orchid Maintenance/Replate Medium
Fig. 1. Particle size analysis of control and treated samples of OMR medium.
3.3. Surface Area Analysis
The surface area of control and treated samples of OMR
medium was investigated using BET method. The control
sample showed a surface area of 2.327 m
/g; however, the
treated sample of OMR medium showed a surface area of
/g. It showed that the surface area was decreased by
3.91% in the treated OMR medium sample as compared to
the control. The decrease in surface area of treated OMR
medium sample may be due to the increase in the particle
size as evident from the particle size analysis. Besides, the
OMR medium has the problem of hygroscopicity  and
the surface area was directly related to the hygroscopic water
content of the sample . Hence, it is assumed that the
treated OMR medium sample with decreased surface area
might reduce the problem of hygroscopicity as compared
with the control sample.
3.4. Thermal Analysis
3.4.1. DSC Analysis
Fig. 2. DSC analysis of control and treated samples of OMR medium.
The DSC thermograms of control and treated samples of
OMR medium are presented in Fig. 2. The thermogram of
control sample showed an endothermic peak at 180.36°C due
to the melting of the sample. In this process, the amount of
heat absorbed (latent heat of fusion, ∆H) was recorded as
23.00 J/g. The similar endothermic peak was observed in
Journal of Plant Sciences 2015; 3(6): 285-293 289
treated sample at 182.58°C and ∆H was recorded as 54.20
J/g. The result of DSC analysis revealed slight alteration
(1.23% increase) in the melting temperature along with
135.7% increase in ∆H. The particle size can influence the
melting temperature and ∆H of the corresponding sample as
it is directly related to the melting properties [28, 29]. Hence,
it might be a reason for the increase in melting temperature
and ∆H of the OMR medium as the particle size was found
increased after the biofield treatment.
3.4.2. TGA/DTG Analysis
The TGA/DTG studies analyse the pattern of thermal
decomposition of the sample during heating. The TGA/DTG
thermograms of the control and treated samples of OMR
medium are presented in Fig. 3. The thermogram of control
sample showed the degradation of the sample in three steps.
Moreover, the first step degradation of the sample was started
at 175.72°C and ended at 203.38°C. Besides, the treated
sample showed two-step degradation, where the first step
commenced at 187.61°C and completed at 237.96°C. It
indicated that the onset temperature of degradation was
increased in the treated sample as compared to the control.
Besides, DTG thermogram data showed that T
observed at 189.19°C in the control sample while 210.41°C
in the treated OMR medium. It indicated that T
increased by approximately 21°C in the treated sample as
compared to the control. Furthermore, the increase in onset
temperature of decomposition and T
in the treated sample
of OMR medium with respect to the control sample may be
correlated with the increased thermal stability. The particle
size has a significant impact on the onset and peak
temperature and they were found directly proportional to
each other . Hence, the increase in particle size of OMR
medium after treatment might be a reason for the increase in
thermal stability. Besides, it is well known that the OMR
medium faces high-temperature treatment (e.g., autoclaving)
before used as culture media where it may suffer from the
problem of thermal degradation. Hence, the treated sample
with increased thermal stability might help in increasing the
efficacy and shelf-life of the treated sample as compared to
Fig. 3. TGA/DTG analysis of control and treated samples of OMR medium.
3.5. CHNSO Analysis
The CHNSO analysis was used to measure the percentage
of elements present in the given sample. The result of
CHNSO analysis of control and treated samples are presented
in Table 2. The data revealed that the percentage of nitrogen
was significantly increased by 22.22% whereas; the
290 Mahendra Kumar Trivedi et al.: Physicochemical Characterization of Biofield
Treated Orchid Maintenance/Replate Medium
percentage of carbon, hydrogen, and oxygen was slightly
decreased as 3.62, 8.51, and 1.92%, respectively in the
treated sample as compared to the control. Besides, the
treated sample showed the presence of sulphur that was not
detected in the control sample. It is well known that nitrogen
is the main component of media that is provided by
ammonium nitrate, potassium nitrate, and peptone. The
increased percentage of nitrogen in the treated sample may
help to improve the growth of orchid culture as compared
with the control.
Table 2. CHNSO data of orchid maintenance / replate medium.
Element Control Treated Percent change
Nitrogen 0.63 0.77 22.22
Carbon 43.90 42.31 -3.62
Hydrogen 6.93 6.34 -8.51
Oxygen 31.18 30.58 -1.92
Sulphur ND 0.27
ND: not detected
3.6. FT-IR Spectroscopic Analysis
Fig. 4. FT-IR spectra of control and treated samples of OMR medium.
The FT-IR spectra of OMR medium (control and treated
samples) are shown in Fig. 4. The sample contains several
ingredients such as ammonium nitrate, disodium EDTA,
ferrous sulphate, potassium nitrate, nicotinic acid, sucrose,
inositol, thiamine hydrochloride, and pyridoxine
hydrochloride, etc. Hence, the major vibration peaks were
observed (Table 3) related to the functional groups present in
these ingredients. The peak at 3336 cm
in the control
sample was assigned to N-H stretching of ammonium nitrate
and O-H stretching (carboxylic acid) due to nicotinic acid
and disodium EDTA  ; however, the broadness of
peak suggests the hydrogen bonding within the compound.
Besides, in the treated sample it was shifted to a lower
frequency at 3323 cm
. Further, the C-H stretching peaks of
disodium EDTA appeared at 2983 and 2896 cm
control sample, whereas, in the treated sample, the peaks
Journal of Plant Sciences 2015; 3(6): 285-293 291
appeared at 2941 and 2898 cm
. The peak due to pyridine
ring of nicotinic acid and pyridoxine HCl was observed at
in the control, while 2833 cm
in the treated
sample. Similarly, the peak at 1753 cm
in the control
sample was assigned to C=O stretching of lactone ring
present in sucrose; however, it was observed at 1735 cm
the treated sample . The peak at 1595 cm
in the control
sample appeared as doublet and it was assigned to ring
stretching of the pyridine ring of nicotinic acid and
pyridoxine HCl . The peak may also merge with the peak
due to S-O bond of CuSO
and P-O bond of KPO
Besides, in the treated sample the corresponding peak was
observed as a singlet at 1608 cm
. Furthermore, the peak at
in the control that was shifted to 1417 cm
treated sample was assigned to N-O symmetric stretching of
and pyrimidine ring of thiamine HCl [31, 32]. The
peaks at 1234 cm
in the control sample and 1236 cm
treated sample was assigned to thiazole ring breathing of the
thiamine HCl . Moreover, the peak at 1146 and 1130 cm
in the control and treated samples, respectively was assigned
to S-O bond in FeSO
, and pyrimidine ring of
thiamine HCl. The peak due to C-O stretching of alcohol
group in pyridoxine HCl was observed at 1047 cm
control and 1058 cm
in the treated sample. The ring
breathing mode of inositol was observed at 1001 cm
control and 999 cm
in the treated sample. Further, the peak
at 858 cm
in both, control and treated sample was assigned
to B-O bond of boric acid and C-H out of plane bending of
thiazole ring in thiamine HCl. The IR peaks of control
sample were well matched with the reported literature. The
FT-IR spectra of the treated sample showed different IR
frequencies of respective functional groups as compared to
the control. It suggested the impact of biofield energy
treatment on the bond strength and dipole moment of the
compounds present in the OMR medium. However, further
studies are needed to analyse the effect of this treatment on
the specific compounds and their functions in OMR medium.
Table 3. Vibration modes observed in orchid maintenance/replate medium.
Functional group Compound Wavenumber (cm
1 O-H stretching,
2 C-H stretching Disodium EDTA,
3 C-H stretching Agar 2815 2833
4 C=O stretching
(lactone) Sucrose 1753 1735
5 Ring stretching
Pyridoxine HCl 1595 1608
9 C-O stretching (C-
OH) Pyridoxine HCl 1047 1058
10 Ring breathing
(carbon ring) Inositol 1001 999
11 B-O stretching Boric acid 858 858
12 Ring deformation
(cycloalkane) Inositol 561 551
3.7. UV-Vis Spectroscopic Analysis
The UV spectra of OMR medium (control and treated
samples) are shown in Fig. 5. The UV spectrum of control
sample showed absorption peaks at λ
equal to 212 and 257
nm. However, the biofield treated sample showed absorption
peaks at λ
equal to 212 and 275 nm. The peak at λ
nm in the control sample was shifted to higher wavelength i.e.
275 nm in the treated sample. It is hypothesized that biofield
energy treatment might affect the HOMO→LUMO transition
within the compounds of OMR medium due to which the peak
257 nm was shifted to 275 nm in the treated sample.
Fig. 5. UV-Vis spectra of control and treated samples of OMR medium.
292 Mahendra Kumar Trivedi et al.: Physicochemical Characterization of Biofield
Treated Orchid Maintenance/Replate Medium
The XRD study showed 10% decrease in the crystallite
size of treated sample along with alteration in the relative
intensities of the peaks. It may occur due to the presence of
microstrains that might be generated after biofield energy
treatment. Moreover, the average particle size and d
increased in treated sample by 20.78% and 39.29%,
respectively as compared to the control. The surface area
data supported the results of particle size analysis and
revealed that the surface area was decreased by 4% in the
treated sample. The increased particle size and reduced
surface area might improve the gelling properties and
reduce the problem of hygroscopicity of the treated sample.
Besides, the melting temperature and ∆H was found
increased in the treated sample as compared to the control.
The TGA results also revealed that the onset temperature of
degradation and maximum degradation temperature was
increased in the treated sample. The increased thermal
stability may help in increasing the efficacy and shelf-life
of the treated sample as compared to the control.
Furthermore, the CHNSO analysis revealed increased
percent of nitrogen along with the presence of sulphur in
the treated sample as compared to the control. The FT-IR
and UV-vis spectra of the treated sample also revealed the
changes as compared to the control. The overall study
revealed the impact of biofield treatment on the physical,
thermal and spectroscopic properties of the OMR medium
that could make it more useful as compared to the control.
Authors greatly acknowledge the support of Trivedi
Science, Trivedi Master Wellness and Trivedi Testimonials in
this research work. The authors would also like to
acknowledge the whole team from the Sophisticated
Analytical Instrument Facility (SAIF), Nagpur and MGV
Pharmacy College, Nashik for providing the instrumental
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