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Raman spectroscopy shows difference in drugs at ultrahigh dilution prepared with stepwise mechanical agitation

DOI:10.51910/ijhdr.v15i1.811
Authors:
Tandra Sarkar at Indian Institute of Engineering Science and Technology, Shibpur
Tandra Sarkar
Atheni Konar at Indian Institute of Engineering Science and Technology, Shibpur
Atheni Konar
Nirmal Chandra Sukul at Visva Bharati University
Nirmal Chandra Sukul
Dipanwita Majumdar
Dipanwita Majumdar
Dipanwita Majumdar
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Objective: The present study aims at deciphering the nature of the water structure of two ultrahigh diluted (UHD) homeopathic drugs by Laser Raman Spectroscopy. Method: Two homeopathic drugs Sulphur and Natrum mur in three UHD 30cH, 200cH and 1000cH were selected for the study. Raman spectra of the drugs and their medium (90% ethanol) were obtained in the wave number region of 2600-3800 cm-1. The intensity ratio at vibration frequencies between 3200 and 3420 (R1) and that between 3620 and 3420 (R2) was calculated for each UHD as well as the control. Results: Raman spectra shows differences in intensities in different UHDs and their control in the stretching vibrations of CH and OH groups. The three UHDs of each drug show an inverse relationship with respect to the R1 values. However, for R2 the relationship of UHD for each drug is positive. Conclusion: R1 provides information about the relative number of OH groups with strong and weak hydrogen bonds. R2 suggests the relative number of OH groups with broken and weak hydrogen bonds. Judged from R1 values the lower is the rank of UHD, the stronger is the H-bond of the OH groups. In the light of R2 values, the higher is the UHD rank the more abundant is the free OH groups. So, hydrogen bond strength and free OH groups together make an effective UHD rank relating to Sulphur and Natrum mur.
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International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
2
Original Article
Raman spectroscopy shows difference in drugs at ultrahigh dilution
prepared with stepwise mechanical agitation
Tandra Sarkar, M.Sc1,3; Atheni Konar, M.Sc1,3; Nirmal Chandra Sukul*, Ph.D1,2; Dipanwita
Majumdar, M.Sc4.; Achintya Singha, Ph.D4; Anirban Sukul, Ph.D1
1Sukul Institute of Homeopathic Research, Santiniketan, West Bengal, India.
2Department of Zoology, Visva-Bharati University, Santiniketan, WB, India.
3Centre for Health Care Science and Technology, IIEST, Shibpur, WB, India.
4Department of Physics, Bose Institute, Kolkata, WB, India.
*Corresponding author. Email: ncsukul@gmail.com
Abstract
Objective: The present study aims at deciphering the nature of the water structure of two ultrahigh
diluted (UHD) homeopathic drugs by Laser Raman Spectroscopy.
Method: Two homeopathic drugs Sulphur and Natrum mur in three UHD 30cH, 200cH and 1000cH
were selected for the study. Raman spectra of the drugs and their medium (90% ethanol) were
obtained in the wave number region of 2600-3800 cm-1. The intensity ratio at vibration frequencies
between 3200 and 3420 (R1) and that between 3620 and 3420 (R2) was calculated for each UHD as
well as the control.
Results: Raman spectra shows differences in intensities in different UHDs and their control in the
stretching vibrations of CH and OH groups. The three UHDs of each drug show an inverse
relationship with respect to the R1 values. However, for R2 the relationship of UHD for each drug is
positive.
Conclusion: R1 provides information about the relative number of OH groups with strong and
weak hydrogen bonds. R2 suggests the relative number of OH groups with broken and weak
hydrogen bonds. Judged from R1 values the lower is the rank of UHD, the stronger is the H-bond of
the OH groups. In the light of R2 values, the higher is the UHD rank the more abundant is the free
OH groups. So, hydrogen bond strength and free OH groups together make an effective UHD rank
relating to Sulphur and Natrum mur.
Keywords: Sulphur, Natrum mur, high dilution, hydrogen bond, free OH.
International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
3
Introduction
Drugs at ultrahigh dilution (UHD) are
frequently used in homeopathy. The
concentration of original drug molecules in
the 12th centesimal dilution reaches to a
dilution beyond 1024, therefore, UHDs
(beyond Avogadro) are likely to be devoid of
original drug molecules. However, the UHDs
show specificity in their biological effects as
reported in various studies. Viewed from the
perspective of their biological function the
UHDs appear different from each other and
also from their diluent medium, in spite of the
fact that their chemical composition is same
as ethanol and water. This paper attempts to
address the pertinent question, “What makes
them different from each other?” using Laser
Raman spectroscopy. Earlier studies using
NMR, electronic, vibrational and Raman
spectroscopy have shown differences in
UHDs1-7.
The difference has been attributed to the
difference in the hydrogen-bonded water
structures. But the exact nature of the water
structures in UHDs has not yet been
elucidated. Recently, an FTIR study by our
group revealed variation in free water
molecules and hydrogen bonding in different
UHD’s8.
The Raman spectroscopy is complementary
to Infrared spectroscopy9. For this, we have
studied Raman spectra of Natrum muriaticum
(Nat-m) and Sulphur (Sulph) in 30cH, 200cH,
and 1000cH potencies with a view to
exploring the differences among them. When
a beam of light passes through a transparent
material a small amount of radiation energy is
scattered. The scattered energy consists of
radiation mostly of incident frequency called
Raleigh scattering. However, certain discrete
frequencies above and below the frequency of
the incident beam are scattered, and this is
referred to as Raman Scattering.
When a sample of molecules, water-ethanol in
UHDs, is subjected to radiation beam of a
particular frequency the electric field
experienced by each molecule varies, and the
induced dipole also undergoes oscillations of
the same frequency. In addition, the molecule
undergoes internal motion such as vibration
or rotation. The vibrational or rotational
oscillation of the molecule would be
superimposed on the oscillating dipole of the
same molecule. This results in changes in the
polarization of the molecule. When a
molecule is put in a static electric field its
positively charged nucleus and negative
electrons are attracted to the negative and
positive pole of the field, respectively. This
causes an induced dipole moment in the
molecule, which is thus polarized.
In this study, we examined the vibrational
Raman Spectra of our samples using laser as a
source of radiation. The laser provides a
narrow monochromatic coherent beam. The
rare gas laser, here Argon, gives intensities
which are one million times greater than that
of sunlight10.
Materials and Methods
Medicines: UHDs of Natrum muriaticum (Nat-
m) and Sulphur (Sulph) in 30cH, 200cH, and
1000cH potencies were used for the
experiments. All drugs, purchased in sealed
vials from the local market, Kolkata, were
products of Dr. Reckeweg, Germany. The
drugs were in 90% ethanol as mentioned on
their label. The control consisted of 90%
ethanol prepared from absolute ethanol by
adding the appropriate volume of High
Performance Liquid Chromatography (HPLC)
water. In this study, we did not use UHDs of
International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
4
alcohol as a second control because UHD
alcohol may have some biological action.
Obviously, a drug cannot serve as a control
for another drug11. Aliquots of the control, as
well as the drugs, were mixed with an
appropriate volume of HPLC water to reduce
the ethanol content to 25%. The optical
density of all the samples was measured by a
UV-VIS spectrophotometer at 200nm and
found to be same.
Raman spectra: Raman measurements were
carried out at 23°C using a micro-Raman
setup (Lab Ram HR, Jobin Yvon) equipped
with an Argon ion laser of wavelength 488nm
and a CCD detector. The spectra were
collected within the wave number region of
2600-3800 cm-1 and analyzed after suitable
baseline correction. The experiments were
repeated thrice and the results were similar.
Here, we have considered the results of the
second experiment.
In water-ethanol solution OH bond undergoes
changes in integral intensity and contour
shape. The intensity ratio of vibrational
frequencies at 3200 and 3420 cm-1(R1)
provides information about the relative
number of OH groups with strong and weak
hydrogen bonds. The intensity ratio of the
modes at 3620 and 3420 cm-1(R2) suggests
the relative number of OH groups with
broken and weak hydrogen bonds12,13. In this
study, we have calculated these two ratios
from the Raman spectra of all the drug
samples and the control used.
Results
Raman spectra in the region of stretching
vibrations of CH (2800-3000cm-1) and OH
(3100-3700cm-1) are presented for different
UHDs of drugs and their aqueous ethanol
control. Fig. 1 shows the spectra of Sulph
30cH and ethanol control, and Fig. 2 Sulph
200cH and 1000cH. In all cases, CH bands of
ethanol have distinct sharp peaks and OH
bands of ethanol and water broad peaks
(Figs.1, 2). The peaks of CH and OH bands of
the drug (Sulph 30cH) and the control and
also of Sulph 200cH and 1000cH with the
same ethanol content 25% show variation in
intensities.
Fig.-1: Raman spectra of Sulph 30cH and aqueous
ethanol control, both in 25% ethanol in the region
of CH and OH stretching vibration.
Fig.-2: Raman spectra of Sulph 200cH and 1000cH
in 25% ethanol in the region of CH and OH
stretching vibration.
International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
5
Intensity Ratio: The intensity ratios R1 and
R2 for control, Sulph 30cH and Nat-m 30cH in
25% ethanol show marked difference from
each other (Figs 3, 4). The ratio values rise
gradually with Sulph 30cH at the bottom and
Nat-m 30cH at the top with the control lying
close to Sulph (Figs.3, 4).
Fig.-3: Intensity ratio I3200/I3420 showing the
difference in Control, Sulph 30cH and Nat-m 30cH.
All in 25% ethanol.
Fig.-4: Intensity ratio I36 20/I3420 showing the
difference in control, Sulph 30cH and Nat–m 30cH.
All in 25% ethanol.
The intensity ratio R1 for control, Sulph 30cH,
Sulph 200cH and Sulph 1000cH is inversely
related to the UHD ranks (Fig.5). The position
of the control is slightly lower than that of
Sulph 30cH (Fig.5).
Fig.-5: Intensity ratio I3200/I3420 showing negative
relation in three potencies of Sulph All in 25%
ethanol.
The intensity ratio R2 for control and three
different UHDs of Sulph rises gradually with
the increase in UHD ranks from the 30th to the
1000th (Fig.6).
Fig.-6: Intensity ratio I3620/I3420 showing positive
relation in three potencies of Sulph. All in 25%
ethanol.
The control lies slightly above Sulph 30cH.
Like Sulph, intensity ratio R1 shows a
negative relation between the three UHD
International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
6
ranks of Nat-m (Fig. 7). The ratio R2 shows a
positive relation with three UHD ranks of
Nat–m (Fig.8) as in Sulph.
Fig.-7: Intensity ratio I3200/I3420 showing negative
relation in three potencies of Nat-m. All in 25%
ethanol. The position of the intensity ratio of the
control is at 1.09.
Fig.-8: Intensity ratio I3620/I3420 showing positive
relation in three potencies of Nat-m. All in 25%
ethanol. The position of the intensity ratio of the
control is at 0.06.
Discussion
The amplitudes of CH and OH stretching
bands increase with the increase in ethanol
content in aqueous ethanol solutions13. In our
study, the amplitudes of CH and OH
stretching bands vary between the control
and the drugs (Fig.1) and in different UHD
ranks of the same drug (Fig. 2) in spite of the
fact that ethanol content is same (25%) in all
the samples. The difference in hydrogen bond
and in bond strength may be responsible for
this.
Intensity Ratio: The intensity ratio R1 is
maximum at 20-25% ethanol concentration
indicating strongest hydrogen bonding of OH
groups of water and ethanol12,13. At this
concentration of ethanol, there occurs
structural rearrangement leading to
stabilization and strengthening of water
structure by ethanol molecules, and
strengthening of hydrogen bonds between
OH groups13.
The ratio R2 decreases as ethanol
concentration increases in water-ethanol
solutions13. The authors hypothesize that
with higher ethanol concentration free OH
groups, not linked with a hydrogen bond,
decreases both in water and ethanol as the
ethanol content increases13. But in our study
Nat-m 30cH has strong hydrogen bond and
maximum free OH groups followed by Sulph
30cH (Figs 3,4). Control is close to Sulph.
Thus, we see that in 25% ethanol the drug,
which has the strongest hydrogen bonding of
the OH groups, also has a maximum number
of free OH groups. Judged from the clinical
perspective Nat-m is a deep and long-acting
drug14,15. Although control and Sulph are close
to each other with respect to the free OH
groups, the latter has stronger hydrogen
bonding of OH groups than the control
(Fig.3). This makes Sulph an effective drug
distinct from the control. It may be
mentioned here that UHDs of aqueous
ethanol control also has some biological effect
International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
7
which could be reduced or eliminated by
dilution with water 1:100016.
In the case of the three UHD ranks of Sulph
and Nat-m, the highest rank 1000cH occupies
the lowest position while the lowest rank 30th
the highest position as evidenced by the ratio
R1 (Figs. 5,7). It means the lower is the rank
the stronger is the H-bond of the OH groups.
But as per the ratio R2, the higher is the rank
the more abundant is the free OH groups
(Figs 6, 8). In homeopathy, the higher is the
UHD rank of a drug the deeper is its
therapeutic effect. For Sulph as well as Nat-m
the deeper efficacy depends more on the free
OH groups rather than on the hydrogen bond
strength of the OH groups. Or, it may depend
on both the factors taken together.
Why do hydrogen bond strength and free OH
groups vary in different UHDs although their
ethanol content is same? The answer lies in
the method of preparation of HD drugs. The
method involves successive dilution of a
mother tincture or a starting material and
mechanical agitation1. Therefore, these two
factors may contribute to the variation in H-
bond strength and free OH groups in UHDs. It
is known that mechanical agitation or
sonication could change the solution
structure in aqueous ethanol, and this change
could produce a biological effect. Haseba et
al17 observed that weak ultra-sonication of
aqueous ethanol solution accelerates the
thermal motion of water molecules in the
solution. This acceleration disperses ethanol
molecules amongst water molecules making
the solution more compact and homogeneous
as compared to the non-sonicated solution.
The authors further observed that sonicated
aqueous ethanol behaves like a mature spirit
being tastier and less toxic than ordinary
alcohol. This means that different UHDs of
drugs might have acquired different levels of
compactness and homogeneity of water
molecules due to different degrees of
mechanical agitation applied. Sonication,
instead of succussion, also makes an effective
UHD1. Therefore, we can say that three
factors like hydrogen bond strength, free OH
groups and compactness cum homogeneity of
water molecules together make an effective
UHD with specificity.
Conclusion
The intensity ratio R1 provides information
about the relative number of OH groups with
strong and weak hydrogen bonds. The ratio
R2 suggests the relative number of OH groups
with broken and weak hydrogen bonds.
Judged from R1 values the lower is the UHD
rank, the stronger is the H-bond of the OH
groups. In the light of R2 values, the higher is
the UHD rank the more abundant is the free
OH groups. So, hydrogen bond strength and
free OH groups together make an effective
UHD relating to Sulphur and Natrum
muriaticum.
Acknowledgements
T.S. and A.K. duly thank Dr. Bholanath
Chakravarty Memorial Trust for providing
research fellowship during the tenure of
work.
Conflict of Interest
None declared.
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Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
8
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International Journal of High Dilution Research 2016; 15(1):2-9
Available online at www.highdilution.org
Cite as: Sarkar T, Konar A, Sukul NC, Majumdar D, Singha A, Sukul A. Raman spectroscopy shows difference in drugs at
ultra-high dilution prepared with stepwise mechanical agitation. Int J High Dilution Res. 2016;15(1): 2-9.
9
Received: Dec 28, 2015. Accepted: March 10, 2016.
© International Journal of High Dilution Research.
Not for commercial purposes.
... In our earlier study, we have demonstrated that these ultrahigh dilutions (UHD) have water structures specific to their UHD rank and original drug. The water structures in UHDs mainly relate to free OH and hydrogen bond strength as evidenced by Raman and vibrational spectroscopy [2][3][4] . UHDs of different drugs are known to produce specific therapeutic effects on the patients depending on their individual symptoms [5][6] . ...
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Article
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... Raman spectroscopy has seldom been used in homeopathy research. 39,40,46 The studies performed by Sarkar et al. 46 and Konar et al. 39 showed an increase in the vibration frequencies of liquid homeopathic potencies, since Raman is able to identify differences between solution and ultradiluted drugs. 47 Our Raman results suggest that the tritura-tion process in the presence of zinc causes changes in the vibration frequencies, which are proportional to the increase in trituration (►Fig. ...
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Article
  • Nov 2021
Background Recent experimental results supporting the dynamization process show modification in the characteristics of solid mixtures. Objective The present work aims to evaluate the physicochemical properties of metallic zinc and lactose, evidencing the interactions between all chemical components presented in dynamized solid mixtures by analytical techniques. Methods Mixtures of zinc and lactose (1:9 w/w) were successively triturated at the same proportion according to the Brazilian Homeopathic Pharmacopoeia, receiving the designation of 10−1 – 10−6 (1dH – 6dH). All samples were submitted to the following characterization techniques: Atomic Absorption Spectrometry (AAS), Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), Differential Scanning Calorimetry (DSC), Thermogravimetry (TG), and Raman Spectroscopy (RS). Results AAS results detected 97.0% of zinc in the raw material, and the triturated zinc lactose system (ZnMet) presented mean values similar to those expected for the physical mixtures: i.e., 9.94%, 1.23%, and 0.11% in the three first proportions (10−1, 10−2, 10−3), respectively. SEM images showed particle size reduction due to the trituration process. The XRD assays of ZnMet 10−3 and 10−6 indicated peak changes at 12.3° and 43.26°, probably associated with modifications of inter-atomic crystalline spacing. The thermal analysis results of dynamized samples suggest modifications in the chemical interaction between zinc and lactose induced by the physical forces applied. RS experiments showed variation in vibration frequencies due to the dynamization procedure, in which marked ZnMet 10−6 spectral modifications were detected at 357, 477, 1086 and 1142 cm−1, and in the wavelength range 860–920 cm−1. Conclusion These results highlight the importance of applying suitable characterization methods to improve our understanding of the properties of homeopathic solid mixtures, whereas the uses of sensitive tools evidence the influence of trituration on the crystalline properties and in the enthalpy variation of dynamized samples.
... [15][16][17][18] Therefore, Raman spectroscopy allows us to understand hydrogen bonding in water and in aqueous solutions in general. [19][20][21] In this study, we established a correlation between the different states of hydrogen bonding of alcohol, which is a vehicle for homeopathic medicine, due to the effect of potentization and measured from Raman data, and the electrical measurement, which provides the ionic state of the liquid. We selected the two homeopathic medicines Chininum purum and Acidum benzoicum for such studies for a specific reason: the former is basic in nature, while the latter is acidic in nature. ...
... To estimate the effect of succussion of alcohol in absence and presence of the medicine on the strength of the stretching OH vibrations, we measured the change in the intensity of OH stretching bands at frequencies 3240 cm À1 , 3420 cm À1 , and 3620 cm À1 , corresponding to strong, weak, and broken hydrogen bonds in water respectively. 19,20 Voltage measurements with 91% alcohol and the two medicines at the three different potencies were made using the electrochemical cell (►Fig. 1A). ...
Investigation of the Origin of Voltage Generation in Potentized Homeopathic Medicine through Raman Spectroscopy
Article
  • Jan 2019
Background For the study of homeopathic medicines in proper perspective, emerging techniques in material science are being used. Vibrational spectroscopy is one such tool for providing information on different states of hydrogen bonding as an effect of potentization. The associated change in electrical properties is also correlated with this effect. Objective From the vibrational spectra, the changes in hydrogen bonding due to dilution followed by unidirectional vigorous shaking (together termed potentization) of 91% ethanol and two homeopathic medicines Chininum purum and Acidum benzoicum have been studied. The aim was to correlate the result with the change in the electrical properties of the system. Methods Raman spectroscopy was used to study the vibrational spectra. A U-shaped glass tube (electrochemical cell), where one arm contained bi-distilled water and the other arm alcohol/homeopathic medicine (the arms being separated by a platinum foil), was used to measure the voltage generated across two symmetrically placed platinum electrodes. Results For all samples, it was observed that potentization affected the intensity of OH stretching bands at the frequencies 3240 cm−1, 3420 cm−1 and 3620 cm−1, corresponding to strong hydrogen bond, weak hydrogen bond and broken hydrogen bond, respectively. With the increase in potency, in the presence and absence of the two medicines in ethanol, the number of OH groups linked by strong hydrogen bonds decreased, while the number of OH groups with weak hydrogen bonds increased. With the increase in potentization, the number of OH groups with broken hydrogen bonds showed a difference in the presence and absence of the medicine. The voltage measurements for ethanol show that, with succussion, the magnitude of voltage increased with the two medicines at lower potencies, but not at higher potency where the voltage is lower. Acidum benzoicum, which is acidic in nature, had higher voltage values (113mV, 130 mV and 118 mV at 6C, 30C and 200C, respectively), compared with Chininum purum, which is basic in nature (20 mV, 85 mV and 65 mV at 6C, 30C and 200C, respectively). Conclusion The experimental results indicate a correlation between the vibrational and electrical properties of the homeopathic medicines Acidum benzoicum and Chininum purum at different potencies.
... But potencies above 12cH, whose dilution is 10 24 , cross the Avogadro number and are, therefore, devoid of original drug molecules. Using FTIR and Raman spec-troscopy and also differential scanning calorimetry (DSC) we have demonstrated that the physical basis of potencies of a drug consists of two factors like hydrogen bond strength of the OH groups and also free OH groups or non-hydrogen bonded water molecules [1][2][3][4][5]. It is hypothesized that potencies initiate their action on cell membrane proteins as in oral mucosa or plant leaves on which the drug is applied [6]. ...
... Water molecules also bind to many parts of a protein nonspecifically. We have already observed that the physical basis of potency consists of two factors, free OH groups and hydrogen bond strength of the OH groups [1][2][3][4][5]. These factors contribute to the specificity of a potency to binding with binding pockets in a protein. ...
High and Ultra Low Concentrations of Sodium Chloride Initiate their Action on Binding Sites of a Protein
Experiment Findings
Full-text available
  • Nov 2017
... No statistics beyond descriptive were reported in all seven experiments (six publications) assessed. 25,53,74,[149][150][151] Further, most experiments used unsuccussed ethanol as a control, thereby leaving out the possibility of unspecific effects due to the succussion procedure. In the present state of the research, there is only weak evidence for specific properties of homeopathic samples identifiable by Raman spectroscopy. ...
Physicochemical Investigations of Homeopathic Preparations: A Systematic Review and Bibliometric Analysis—Part 3
Article
  • Oct 2020
Objectives: In parts I and II of our review of physicochemical research performed on homeopathic preparations, we identified relevant publications and analyzed the data in terms of individual experiments, looking for the most promising techniques that were used in the past. In this third part, we analyze the results of the experiments seeking to extract information about the possible modes of action underpinning homeopathic preparations. Methods: We summarized the results from the 11 experimental areas previously introduced, extracting the general findings and trends. We also summarized the results in terms of specific research topics: aging, medium used for potentization, sample volume, temperature, material of potentization vessel, and, finally, the use of molecules to probe homeopathic samples. Results: We identified a number of effects that appear consistently throughout the data: Differences to controls seem to increase with: time, moderate temperature, small samples volume, and in ionic medium, whereas high temperatures seem to abolish differences to controls. Based on the present analysis, there is no consistent evidence to date for the nanoparticle hypothesis to explain specific homeopathic treatment effects. However, the quantum coherence domain hypothesis, the dynamic water cluster hypothesis, and the weak quantum theory are still contenders and need to be further assessed experimentally. Conclusions: The field requires further targeted experimentation to validate past findings reporting differences between homeopathic dilutions and controls, and to expand these findings by specifically testing the three main working hypotheses that are currently at hand.
... In the present experiment we used a potency of Merc cor and ordinary ethanol without dynamization. So dynamization might have added to the strength of hydrogen bonds in Merc cor 200 cH due to mechanical agitation during the preparation of the potency (Sarkar et al. 2016). Hydrogen bond is actually an electrostatic attraction between 2 close water molecules. ...
Free and Bound Water in Three Different Concentrations of a Homeopathic Drug Mercurius corrosivus 200 cH and its Vehicle Ethanol
Article
Full-text available
  • Feb 2019
Mercurius corrosivus 200 cH, prepared by successive dilution followed by succussion and ethanol without succussion are both in 90% ethanol. Each of them was diluted first with water 1 : 1000 and then each dilution as a unit was further diluted with appropriate volume of water to make 0.5, 2 and 4% solution. Each of these diluted samples of Merc cor/ethanol was mixed separately with lactose in the proportion of 50 μl per 1g lactose. One lactose sample was mixed with water alone at 50 μl per 1g lactose. All the lactose samples were separately tested by thermogravimetry (TG) to assess the amount of free and bound water. Each test sample was heated from 30 o C to 200 o C at the rate of 10 o C per min. While TG curve indicates mass loss (%) as a function of temperature , differential thermogravimetry (DTG) shows rate of change of mass loss in relation to temperature. Lactose samples containing 0.5, 2 and 4% ethanol unit have 7.7, 10.7 and 7.9% free water, respectively. Lactose samples with 0.5, 2 and 4% Merc cor 200 cH unit have 12.3, 5.2 and 8.3% free water, respectively. Lactose sample with water (1 : 50µl) has 3.5% free water at 36.4°C temperature and 5% bound water at 147.7°C. Bound water varies between 3.5 and 4.6% in both ethanol and Merc cor samples at a temperature of 146-147.7 o C. Loss of free water in both Merc cor and ethanol occurred between 45.3-63.3°C. There is no linearity in relationship between free water and total water content and between thermal energy needed to remove free water and the total water content in all the test samples.
Raman Spectroscopy of High Dilutions of Two Drugs in Aqueous Ethanol Solution Shows Variation in Clathrate Hydrate Crystals
Article
Full-text available
  • Feb 2022
Physicochemical Investigations of Homeopathic Preparations: A Systematic Review and Bibliometric Analysis—Part 2
Article
  • Jul 2019
Objectives: In Part 1 of the review of physicochemical research performed on homeopathic preparations the authors identified relevant publications of sufficient reporting quality for further in-depth analysis. In this article, the authors analyze these publications to identify any empirical evidence for specific physicochemical properties of homeopathic preparations and to identify most promising experimental techniques for future studies. Methods: After an update of the literature search up to 2018, the authors analyzed all publications in terms of individual experiments. They extracted information regarding methodological criteria such as blinding, randomization, statistics, controls, sample preparation, and replications, as well as regarding experimental design and measurement methods applied. Scores were developed to identify experimental techniques with most reliable outcomes. Results: The publications analyzed described 203 experiments. Less than 25% used blinding and/or randomization, and about one third used adequate controls to identify specific effects of homeopathic preparations. The most promising techniques used so far are nuclear magnetic resonance (NMR) relaxation, optical spectroscopy, and electrical impedance measurements. In these three areas, several sets of replicated high-quality experiments provide evidence for specific physicochemical properties of homeopathic preparations. Conclusions: The authors uncovered a number of promising experimental techniques that warrant replication to assess the reported physicochemical properties of homeopathic preparations compared with controls. They further discuss a range of experimental aspects that highlight the many factors that need to be taken into consideration when performing basic research into homeopathic potentization. For future experiments, the authors generally recommend using succussed (vigorously shaken) controls, or comparing different homeopathic preparations with each other to reliably identify any specific physicochemical properties.
Vibrational and Raman spectroscopy provide further evidence in support of free OH groups and hydrogen bond strength underlying difference in two more drugs at ultrahigh dilutions
Article
Full-text available
  • Jan 2016
Objective: To confirm that free water molecules and hydrogen bond strength of OH groups underlie difference between two homeopathic drugs at ultrahigh dilution (UHD). Method: FTIR and Laser Raman spectra of UHDs of X-ray and Magnetis Poli Ambo were obtained in the wave number regions of 2400-4000 cm⁻¹ and 2400-4200 cm⁻¹, respectively. Mother tincture (MT) were prepared by exposing ethanol water to X-radiation for X-ray and magnetic field for Magnetis. Spectra of the reference water and the three UHDs of Ethanol were also taken. All the samples were in water-ethanol solution in which the ethanol content was 25%. For FTIR the difference spectrum (absorbance of a UHD minus absorbance of reference water) was obtained after normalization of the spectrum at 3410 cm⁻¹. For Raman spectra the intensity ratio at vibration frequencies between 3200 and 3420 cm⁻¹ (R1), and that between 3620 and 3420 cm⁻¹ (R2), were calculated for each UHD. The intensity at 3600 cm⁻¹ in the difference spectra (FTIR) represents the number of free water molecules in UHDs. R2 values in Raman scattering suggest the same thing. Results: The data in both cases follow almost a similar pattern of difference among the UHDs studied here. For example, X-ray: FTIR 14 < MT < 32 < 8, Raman: 14 < 8 < 32 < MT. Magnetis: FTIR: 14 < MT < 8 < 32, Raman:14 < 8 < 32 < MT. R1 values, which indicate hydrogen bond strength in the OH groups, differ from each other in the UHDs and MTs studied. Conclusion: The UHDs and MTs differ from each other with respect to the two factors like free water molecules and hydrogen bond strength.
Manifestation of hydrogen bonds of aqueous ethanol solutions in the Raman scattering spectra
Article
Full-text available
  • Mar 2011
Spectra of Raman scattering of light by aqueous ethanol solutions in the range of concentrations from pure water to 96% alcohol are studied. For water, 25%, and 40% solutions of ethanol in water, as well as for 96% alcohol the Raman spectra are measured at temperatures from the freezing point to nearly the boiling point. The changes in the shape of the stretching OH band are interpreted in terms of strengthening or weakening of hydrogen bonds between the molecules in the solution. The strongest hydrogen bonding of hydroxyl groups is observed at the ethanol content from 20 to 25 volume percent, which is explained by formation of ethanol hydrates of a definite type at the mentioned concentrations of alcohol. This is confirmed by means of the method of multivariate curve resolution, used to analyse the Raman spectra of aqueous ethanol solutions. With growing temperature the weakening of hydrogen bonding occurs in all studied systems, which consists in reducing the number of OH groups, linked by strong hydrogen bonds. (laser applications and other problems in quantum electronics)
Differences in Median Ultraviolet Light Transmissions of Serial Homeopathic Dilutions of Copper Sulfate, Hypericum perforatum, and Sulfur
Article
Full-text available
Homeopathic remedies are produced by potentising, that is, the serial logarithmic dilution and succussion of a mother tincture. Techniques like ultraviolet spectroscopy, nuclear magnetic resonance, calorimetry, or thermoluminescence have been used to investigate their physical properties. In this study, homeopathic centesimal (c) potencies (6c to 30c) of copper sulfate, Hypericum perforatum, and sulfur as well as succussed water controls were prepared. Samples of these preparations were exposed to external physical factors like heat, pressure, ultraviolet radiation, or electromagnetic fields to mimic possible everyday storage conditions. The median transmissions from 190 nm to 340 nm and 220 nm to 340 nm were determined by ultraviolet light spectroscopy on five measurement days distributed over several months. Transmissions of controls and potencies of sulfur differed significantly on two of five measurement days and after exposure to physical factors. Transmissions of potencies exposed to ultraviolet light and unexposed potencies of copper sulfate and Hypericum perforatum differed significantly. Potency levels 6c to 30c were also compared, and wavelike patterns of higher and lower transmissions were found. The Kruskal-Wallis test yielded significant differences for the potency levels of all three substances. Aiming at understanding the physical properties of homeopathic preparations, this study confirmed and expanded the findings of previous studies.
Raman and IR spectroscopy research on hydrogen bonding in water–ethanol systems
Article
Full-text available
  • Sep 2010
Vibrational spectroscopy provides invaluable information about hydrogen bonding in aqueous solutions. To study changes in H-bonding due to increase of ethanol concentration in water, we perform research on water–ethanol binary mixtures with various mixing ratios using a combination of Raman scattering and IR absorption techniques. We study Raman spectra from 200 to 4000 cm−1 excited at 488 nm and IR spectra from 500 and 4000 cm−1 for solutions with different ethanol concentrations from pure water to pure ethanol. Using the intensity ratio of OH stretching band taken at 3200 and 3420 cm−1 for Raman spectra and at 3240 and 3360 cm−1 for IR spectra we evaluate the strength of H-bonding. Maximal strength of H-bonding in water–ethanol mixture corresponds to ethanol concentration 15–20% w/w. We explain it by the presence of transient ethanol hydrates similar in composition to gaseous clathrates with stoichiometric water/ethanol ratio 5:1. Further weakening of H-bonding with ethanol concentration is caused by the formation of chain aggregates from ethanol/water molecules. In addition, we apply other approaches, such as multivariate curve resolution-alternating least squares analysis, decomposition of water Raman stretching band, and comparison of water Raman stretching band in ethanol solutions to that of gas clathrates to support this hypothesis.
High-field 1H T1 and T2 NMR relaxation time measurements of H2O in homeopathic preparations of quartz, sulfur, and copper sulfate
Article
Full-text available
Quantitative meta-analyses of randomized clinical trials investigating the specific therapeutic efficacy of homeopathic remedies yielded statistically significant differences compared to placebo. Since the remedies used contained mostly only very low concentrations of pharmacologically active compounds, these effects cannot be accounted for within the framework of current pharmacology. Theories to explain clinical effects of homeopathic remedies are partially based upon changes in diluent structure. To investigate the latter, we measured for the first time high-field (600/500 MHz) 1HT(1) and T(2) nuclear magnetic resonance relaxation times of H2O in homeopathic preparations with concurrent contamination control by inductively coupled plasma mass spectrometry (ICP-MS). Homeopathic preparations of quartz (10c-30c, n = 21, corresponding to iterative dilutions of 100(-10)-100(-30)), sulfur (13x-30x, n = 18, 10(-13)-10(-30)), and copper sulfate (11c-30c, n = 20, 100(-11)-100(-30)) were compared to n = 10 independent controls each (analogously agitated dilution medium) in randomized and blinded experiments. In none of the samples, the concentration of any element analyzed by ICP-MS exceeded 10 ppb. In the first measurement series (600 MHz), there was a significant increase in T(1) for all samples as a function of time, and there were no significant differences between homeopathic potencies and controls. In the second measurement series (500 MHz) 1 year after preparation, we observed statistically significant increased T(1) relaxation times for homeopathic sulfur preparations compared to controls. Fifteen out of 18 correlations between sample triplicates were higher for controls than for homeopathic preparations. No conclusive explanation for these phenomena can be given at present. Possible hypotheses involve differential leaching from the measurement vessel walls or a change in water molecule dynamics, i.e., in rotational correlation time and/or diffusion. Homeopathic preparations thus may exhibit specific physicochemical properties that need to be determined in detail in future investigations.
Research on Homeopathy: State of the Art
Article
Full-text available
  • Nov 2005
In this paper, we review research on homeopathy from four perspectives, focusing on reviews and some landmark studies. These perspectives are laboratory studies, clinical trials, observational studies, and theoretical work. In laboratory models, numerous effects and anomalies have been reported. However, no single model has been sufficiently widely replicated. Instead, researchers have focused on ever-new models and experiments, leaving the picture of scattered anomalies without coherence. Basic research, trying to elucidate a purported difference between homeopathic remedies and control solutions has also produced some encouraging results, but again, series of independent replications are missing. While there are nearly 200 reports on clinical trials, few series have been conducted for single conditions. Some of these series document clinically useful effects and differences against placebo and some series do not. Observational research into uncontrolled homeopathic practice documents consistently strong therapeutic effects and sustained satisfaction in patients. We suggest that this scattered picture has to do with the fourth line of research: lack of a good theory. Some of the extant theoretical models are reviewed, including placebo, water structure, silica contamination, energy models, and entanglement models. It emerges that local models, suggesting some change in structure in the solvent, are far from convincing. The nonlocal models proposed would predict that it is impossible to nail down homeopathic effects with direct experimental testing and this places homeopathy in a scientific dilemma. We close with some suggestions for potentially fruitful research.
Variation in free and bound water molecules in different homeopathic potencies as revealed by their Fourier Transform Infrared Spectroscopy (FTIR)
Article
  • Jan 2014
Homeopathic potencies 12cH and above cross the Avogadro number, and as such do not contain any original drug molecules in their aqueous ethanol medium. It is thought H-bonded water structures preserved by ethanol carry the information of initial drug molecules. Potentized drugs show some differences with respect to their infrared (IR) absorption spectra. In a water-ethanol solution, free water molecules vary according to the concentration of ethanol. In the present study the concentration of ethanol has been kept constant at 0.03 molar fractions in 6 different homeopathic potencies. Aim: to see whether different homeopathic potencies having fixed ethanol content show variation in FTIR spectra and also free water molecules. Two potencies like 8cH and 32cH of three homeopathic drugs Natrum mur, Cantharis and Nux vomica were used in the study, and their ethanol concentration was kept fixed at 0.03 molar fraction. The control was considered to be aqueous ethanol at the same concentration. Spectrum of reference water was also taken. Fourier transform infrared (FTIR) absorption spectra were obtained in the wave number region of 4000 - 2800 cm-1. The half-width at half-maximum was measured for each spectrum. The intensity of each spectrum was normalized at 3410 cm-1 close to the peak. The difference spectrum (absorbance of drug solution - absorbance of reference water) for each drug and the control was obtained. FTIR spectra showed variation in absorbance intensity on both the high and low frequency side of the O-H stretching band in different drugs as well as the control. The C-H stretching band of 2977 cm-1 also showed variation in intensity in different drugs. In the difference spectra the absorbance intensity at the dip at 3630 cm-1 varied in different drugs and the control. The decrease in intensity at 3630 cm-1 and subsequent rise in intensity at lower frequency region represent the level of free water molecules and strong alcoholic O-H band around 3250 cm-1, respectively. The drug and the control solutions show distinct variation in their FTIR spectra. The drugs have different levels of bound and free water molecules although their ethanol concentration is same.
High Dilution Effects: Physical and Biochemical Basis
Article
  • Jan 2005
Since the subject of high dilution effects is still a subject for debate, this volume provides evidence in support of effects from control clinical studies, clinical records from veteran physicians, controlled experiments on animals and plants, and in vitro tests without any organisms (Chapter II). An overview of the methods for preparing drugs at ultra high dilution is also provided as well as the basic principles of homeopathy, which has been alleviating human suffering through the use of these drugs for several hundred years (Chapter I). Chapter III provides physical basis of high dilutions as evidence from the NMR, IR, UV and fluorescence spectra of those drugs. Since water is used as the diluents media, the structure and dynamics of water polymers in relation to high dilution are discussed in order to facilitate easy comprehension of this physical aspect, the basic principles of spectroscopy are also described. Chapter IV focuses on the mechanism of action of potentized drugs in the living system, discussing the structure of the cell, the plasma membrane, the integral proteins on the membrane, the interaction between these proteins and high dilutions and the manifestations of the therapeutic effects of high dilutions. Some aspects, peculiar to homeopathy, such as the chief miasm psora, and the literalities and time modalities of symptoms and drug action are interpreted from a scientific perspective. Chapter IV ends with a brief discussion on water structures and the origin of life to show the natural evolution of high dilution effects. The book not only helps in understanding the physical basis of high dilutions and their mechanism of action in organisms but provides many new avenues of investigation into this interdisciplinary field of science. © 2005 Springer Science + Business Media, Inc. All rights reserved.
Sepia 200 cH in 1:1,000 dilution counteracts the effect of salt stress in cowpea seedlings but vehicle 90% ethanol proves ineffective in the same dilution
Article
  • Jan 2012
Background: Soil salinity severely affects crop yield all over the world. In a recent study, we found that homeopathic drug Natrum muriaticum 200 cH improved growth in germinating cowpea [Vigna unguiculata (L.) Walp] seeds. Aim: In the present study, we tested homeopathic drug Sepia succus, which is complementary to Nat-m on cowpea seedlings under salt stress. Methods: Cowpea seedlings grown over moist filter paper in Petri dishes were divided in 4 groups: (1) control in sterile water, (2) in 50 mM NaCl solution, (3) seeds pretreated with 90% ethanol diluted with water 1:100 and then transferred to 50 mM NaCl solution, and (4) seeds pretreated with Sep 200 cH diluted with water 1:100 and transferred to 50 mM NaCl solution. In another experiment the groups were same, but the dilution of 90% ethanol and Sep 200 cH was 1:1,000 instead of 1:100 to further reduce the ethanol content in both drug and its vehicle 90% ethanol, to minimize or abolish the effect of alcohol. The data were analyzed by ANOVA followed by Student's t-test. Results: Sep 200 cH in both 1:100 and 1:1,000 dilutions significantly increased the growth, sugar, chlorophyll, protein and water content of seedlings as compared to the untreated salt-stressed group. The effect with the 1000th dilution of Sep 200 cH was more pronounced compared to the 100th dilution. Vehicle 90% ethanol in 1:100 dilution produced some positive effects on the seedlings, but its 1000th dilution produced no such effect. Conclusions: Sep 200 cH counteracted the effects of salt stress in cowpea seedlings, and its 1000th dilution was more effective than the 100th dilution. The effect of alcohol was totally eliminated with the 1000th dilution of 90% ethanol. Therefore, the 1000th dilution could retain the drug effect and eliminate the vehicle effect.
Fundamentals of Molecular Spectroscopy
Article
  • Jan 1966
Variation in Fourier Transform Infrared Spectra of Some Homeopathic Potencies and Their Diluent Media
Article
  • Nov 2005
The aim of this study was to determine whether potentized homeopathic drugs and their diluent media differ from each other with respect to their Fourier transform infrared (FTIR) spectra. FTIR spectra of Nux vomica 30C, Lycopodium 30C, Santonin 30C, Cina 30C, Cina 206C, Cina 1006C, and their diluent media 90% ethanol and Ethanol 30C were obtained in the wave number range of 2000-1000 cm1 at 20 degrees C. Potassium bromide powder soaked with the potencies, pressed into pellets, and air dried were used to measure the spectra. Because water structures in homeopathic potencies are thought to carry specific information on drug molecules and because O-H bending vibrational band (v2) exclusively belongs to water, the study was restricted to the bands in that wave number region. Alcohol has no absorption in the O-H bending region. The potencies were found to differ from each other and their diluent media in the number of v2 bands, their wave number (cm1), shape, and half-width (cm1) of the bands. The number and other characteristics of the v2 band represent the number of hydrogen-bonded water species and their hydrogen-bonding strength, respectively. The potencies and their diluent media therefore differ from each other in the number of hydrogen-bonded water species and their hydrogen-bonding strength. The observation that KBr pellets soaked with a potentized drug retains its specific spectral absorption properties simply confirms that medicated sucrose globules, used in homeopathic dispensing, are capable of retaining the therapeutic properties of the drug.