Pulsatilla pratensis Anhang
http://ir.dut.ac.za/bitstream/handle/10321/324/Erasmus_2004.pdf?sequence=1&isAllowed=y
[Fourie Erasmus]
A comparative study of the NMR spectra of parallel potencies of Pulsatilla pratensis, prepared
according to Hahnemannian and Anthroposophical
Extended Medicine methods.
The purpose of this study was to analyse and compare the NMR spectra of
three analogous ultra-high dilutions, prepared according to the classical Hahnemannian method
and two Anthroposophical methods (Wala and Weleda).
Comparison was made in terms of the chemical shift and relative
integration values of the CH2, CH3, H2O and OH signals obtained via NMR
spectroscopic methods.
All three samples analysed were theoretically identical, except for the
method of dynamization specific to each method.
Comparison was drawn between all three methods. It was hypothesized that
the method of dynamization plays a significant role
in the potentization process and therefore
in the establishing of the physico-chemical
identity of each individual sample. This in turn could indicate a
pharmacological uniqueness in each sample.
The experiment was conducted as per the limitations of the scientific
method.
All samples were diluted and potentized to D12
potency level. Samples were produced in 16ml volumes and transported to the NMR
spectroscopy laboratory at the University of Kwa Zulu
Natal, Pietermaritzburg.
During the last forty years, NMR spectroscopy has been utilized to an
increasing degree in researching the physico-chemical
composition of homoeopathic preparations.
In particular, NMR appears to be a useful experimental technique in a
continuing attempt to examine proposed theories as to the mechanisms present in
the production of homoeopathic remedies (Weingartner:1990)
A Varian 500MHz INOVA spectrometer was used in the experiment, having a
5mm broadband switchable probe and a 5mm inverse detection probe. The III pulse
angle
was set at 90° and the temperature maintained at 25° C. A volume of 1.75
ml of each sample was drawn into a coaxial tube by means of a new micropipette.
Acetone was
used as an external lock and ethanol as the reference. NMR spectroscopic
analysis was conducted on three samples of each group.
The data was recorded and represented in the form of NMR spectra,
providing chemical shift and integration values for the CH2, CH3, H2O and OH
peaks.
All three sample groups were statistically compared by application of
the Pairwise Comparison test to the chemical shifts
and the relative integration values. The level of significance was set at
α = 0.05 for all test comparisons.
The results of this study showed that statistically significant
differences were observedon NMR spectroscopy between Hahnemannian and Anthroposophical
potentising methodologies, most notably in the
chemical shift of the CH2 and CH3 signals.
These differences would make their equivalent use in the pharmacological
industry questionable. This study also served to validate the use of NMR
spectroscopy as a valuable tool in researching ultra-high dilutions.
The homoeopathic process of preparing medicine was introduced by
Hahnemann in the fifth edition of the Organon, in
1831. It is characterized by four distinguishing features (Gaier
1981:456):
1. It is a purely mechanical and mathematico-physical process.
2. The procedure involves neither uncertain,
unreliable nor immeasurable factors.
3. The resultant product is stable and can
readily be maintained that way.
4. The process is theoretically illimitable,
though it becomes laboriously time consuming in the higher range of potencies.
Potentized substances possess certain
attributes:
1. Quantitative (chemical) reduction linked to
qualitative increment of therapeutic (reactive) property.
2. Physical solubility (even of substances,
like metals, believed to have been insoluble).
3. Physiological assimilability and
bioavailability.
4. Altered therapeutic activity (suppression of
primary (direct), and enhancement of secondary (reactive) effect of drugs.
In homoeopathic potentization three scales are
used:
1. The decimal scale where the first potency
contains one tenth part of the crude substance and each succeeding potency
contains one tenth part of the potency immediately preceding. The decimal
potency is indicated by the numerals denoting the deconcentration
with the suffix D or X.
2. The centesimal scale where the first potency
contains one hundredth part of the crude substance and each succeeding potency
contains one one-hundredth part of the potency immediately preceding. The
decimal potency is indicated by the numerals denoting the deconcentration
with the suffix C.
3. The quinquagenimillesimal
(50-millesimal) scale, involving a different method of preparation altogether,
resulting in each potency level containing one fifty thousandth of the
preceding level.
For the purposes of this study the Hahnemannian
sample was serially diluted on a 1:10 scale with 100 succussions
between each dilution.
ANTHROPOSOPHICAL EXTENDED MEDICINE
Anthroposophical extended medicine is one of a
number of practical applications of the work of the Austrian scientist and
philosopher, Rudolf Steiner (1861-1925), the founder of Anthroposophy.
Anthroposophy seeks to extend the understanding of man in a perspective
exceeding purely material or mechanical means. It is a philosophical
and scientific approach to creation, emerging from, but extending beyond
the foundations of natural science. In particular, it rejects the reductionist
approach and recognizes man as consisting of not only the physical body, but
also the soul and spirit (Evans & Rodger 1992:10).
The application of the principles of Anthroposophy has reached far
beyond the purely philosophical. Anthroposophical
principles have been successfully applied to various fields, resulting in an
innovative approach to not only medicine, but also new forms of education, art,
architecture, caring for the handicapped, agriculture and economics (Evans
& Rodger 1992:10).
Anthroposophical medicine came about as a result of
a group of medical doctors recognizing that this extended physiology, of
regarding man not only as a physical body,
but as an integrated four-fold being, had remarkable implications for
medical treatment.
Through Steiner’s collaboration with a Dutch doctor, Ita
Wegman, the foundation for a new approach to medicine
was laid. Their collaboration resulted in a book for the medical profession, „Fundamentals
of Therapy’, and the first Anthroposophical clinic was
opened at Arlesheim, Switzerland. As Steiner was not
a medical doctor, he worked with qualified practitioners in the development of anthroposophical medicine. He insisted that it should extendorthodox medicine rather than become an alternative.
Thus all anthroposophical practitioners
qualify first in conventional medicine, thereafter do further study in the
understanding of man in health and illness from the anthroposophical
perspective (Evans & Rodger 1992:10).
The aim of anthroposophical medicine is to
stimulate the natural healing forces in the patient. These are the life forces
which maintain the physical body and opposes decay. Steiner describes man in
terms of this life force, as a four-foldbeing (Evans
& Rodger 1992:21).
The physical body,
The etheric body.
Comprised of non-physical formative forces, particularly active in growth and
nutrition.
The astral body. Expressing itself particularly
in the nervous system.
The ego. Representing man’s spiritual core and
self-consciousness.
This is expressed in the muscular activity and the blood.
Anthroposophical medicine thus seeks to understand
illness in holistic terms, based on the way these four aspects of man
interrelate to form a whole.
Anthroposophical medicine consists of two distinct
productions: Wala and Weleda.
Potencies most often employed are produced in a 1:10 dilution ratio and lie
within the decimal scale.
Dynamization however, differs significantly from
the Hahnemannian method.
The Weleda succession technique requires that
the container be moved in an oscillatory motion of a figure eight (the sign of
infinity)
1. Wala preparations undergo dynamization by a swift, horizontal movement of the arm
from back to front, resulting in a vortex being created within the container
2. Anthroposophical potentising
continues for a period of two and a half minutes for plant substances and four
minutes for metals, whereafter the liquid is allowed
to settle until all movement ceases.
Each respective action; the completion of one period oscillation and one
period vortex creation, is considered to be equivalent to one period of dynamization in the Hahnemannian
method.
THE ROLE OF POTENTIZATION IN HOMOEOPATHY
By experimental evidence, the effect of homoeopathic preparations in succussed high dilutions on a living organism is no longer
anecdotal. Positive results in studies on cellular elements, plants and animals
disprove the possibility of a simple placebo effect (Smith &
Boericke:1968).
Still, the mechanism of succussed high
dilutions and its action on an organic system remains undecided. Most theories
endorse a scheme of some physical restructuring of the solvent, as a result of
both serial dilution and succussion of the substance
(Anagnostatos:1991; Barnard:1965; Smith & Boericke:1968).
Suggested theories commonly focus on complex organised hydrogen bonded
molecules in ethanol-water mixtures, or electromagnetic coherence and resonance
phenomena.
In an attempt to understand the mechanism of succussed
high dilutions, the divergence from a causal, biochemical model is necessary,
as much of succussed high dilution medicinal
substances fall beyond Avogadro’s limit, where theoretically no original solute
is present in the substance. Even below this limit (D24), the chemical
bioavailability
is usually too insignificant to produce a biochemical effect on the
physical body, or in fact, to easily justify a causal effect within an orthodox
scientific paradigm.
Although investigations within current scientific paradigms are
essential, a non-reductionist approach as suggested by Wallach (2000) may
afford a better opportunity
to understanding the phenomenon.
Barnard and Stephenson first hypothesized that it is not the solute but
the structure of the solvent that is the active participant, and thus the
phenomena of interest in succussed high dilutions,
since many remedies are diluted to such an extent that there is theoretically
none of the original solute remaining in the remedy. They postulated the hypothesis
of stereospecific solvent molecule polymers formed by
association with the original solute (Sacks:1983).
These polymers would self-replicate during the process of serial
dilution and succussion. Addition of monomers in a
specific pattern occurs until a certain length is reached, whereafter
it is broken by the shearing force of the applied succussion.
New, shorter polymers lengthen in the same manner until maximum dimensions are
reached.
The process would repeat itself throughout the dilution and succussion process. These polymers are deemed to be the
informational molecules which are “recognized” by biological systems. (Anagnostatos et al.’s (1991) model of succussed
dilutions centred on the concept of clathrates. He
hypothesised the specific organization of molecules
of the solvent in homoeopathic microdilutions
which can maintain the properties of an initial substance not effectively
present (Anagnostatos et al.:1991).
This is based on the idea of the formation of shells of organised
hydrogen-bonded molecules of the solvent (clathrates)
around aggregates of a small number of molecules
of base substance.
Together with different inertial properties, the succussion
forces clusters of base molecules out of their clathrates,
with new clathrates forming around them. The
displaced clathrate leaves a hollow in the matrix, a
“core clathrate”, and a “mantle” forms around this
core. At the point where no base substance is present, the application of succussive force results in core clathrates
moving out of mantle clathrates and stimulate the
formation of new clathrates. This process is
perpetuated to result in a specific molecular matrix, bearing the informational
imprint of the original substance (Ross 1997:8).
The work of Resch and Gutmann
(1991) pointed to a highly organised structure inherent in water which is able
to be substance-specifically modified by interaction with an added substance or
solute.
It was proposed that a “super molecular system” forms within succussed solutions. This is distinguishable from normal
liquid water through “solvation spheres” or
“hydration shells” forming around hydrophilic molecules, and a network of
“inner surface” molecules at the interface of hydrophobic molecules.
Hydrophobic molecules within a liquid may adopt structural information from the
added substance. This would be preserved within its oscillating expression and,
in turn, exhibit a strong influence on the oscillating pattern of the liquid as
a whole.
The dilution process results in an interface between the solute and the
solvent, which allows for the transfer and integration of the structural
information content into the
new dilution.
Berezin (1991) presented a model based on
isotopic diversity. He proposes a model of homoeopathic action centred on the
patterning of stable isotopes in water.
This argument is based on the notion that the succussion
process results in a non-equilibrium state within the liquid, with an excess of
free energy. This would make for a system vulnerable to pattern formation.
Dissolved molecules would be able to cause re-ordering and positional
arrangement of isotopes within water, water having three isotopic degrees of
freedom; H to D and 170 or 180 to 160.
As a change of a singular neutron in a substance with atomic mass 200
could cause a variation of 0,5%, it would follow that such isotopic change
could cause substantial variations in atomic vibrational
frequencies, bond strengths and changes in chemical activity. Isotopic
combinations provide immense information storage capability.
Fragments would be sufficient to provide the structural information
requirements to a next stage dilution. An example of such a degenerate system
is that of crystallisation where a „micro-change’ in the lattice structure will
result in an ordered structure formation conducive to that change throughout
the rest of the crystal.
Current theories on the mechanism of homoeopathy more and more demand
the ability of lateral thinking. Within quantum theory the opportunity may have
appeared
whereby the link between consciousness and physical matter may move from
a pseudo-scientific regard into the domain of true science. The development of
this notion
has been expanded from the works of Bohm,
Schrödinger and Bohr amongst others (Davies &Brown 1986:32).
In spite of rigorous care and precision, scientific research in
homoeopathy tends to show unrepeatable and anomalous results. It has been
suggested that this may not be completely independent of and uninfluenced by
the researcher. The „Pauli–effect’ is a simple example of the observer as
unintentional participant in a scientific experiment (McEvoy
& Zarate 1991:96).
Robert Oppenheimer stated that “the physical world is not completely
determinate. There are predictions you can make about it but they are
statistical; and any event has in
it the nature of a surprise, of the miracle, of something that you could
not figure out. Physics is predictable - but within limits; its world is
ordered but not completely causal”.
He also remarks that every atomic event is individual. It is not, in its
essentials, reproducible” (Whitmont 1991:4).
Wallach (2000) proposed a non-local interpretation of the homoeopathic
phenomenon. He suggests that a more precise explanation of the mechanism of
homoeopathy is
more likely to be found in conjectures made around concepts based on
quantum theory, rather than theorizing within a purely physico-chemical
paradigm.
The concept on non-locality is perhaps best illustrated in the EPR
paradox.
According to the Copenhagen interpretation presented by Bohr, the
existence of an external world independent of an observer is problematic. One
is in effect, unable to
solve the problem of how the universe exists without an observer looking
at it. Dealing with phenomena, appearances and regularities in phenomena, he
essentially claims
that reality is ultimately ambiguous and unspecifiable,
as affirmed in the EPR paradox. Herein Bohr refuted Einstein’s locality
principle of separateness of phenomena.
Bohr basically states that quantum mechanics does not permit a
separation between the observer and the observed. Any observed phenomenon (in
the case of the EPR
thought experiment; the two electrons) and the observer are part of a
single system – independent of distance and the speed of light, and therefore,
time. It has thus been stated that the EPR experiment does not demonstrate the
incompleteness of quantum theory, but the naiveté of assuming local conditions
in atomic systems. Once they
have been connected, atomic systems are neverseparate
(McEvoy & Zarate
1991:166).
Walach developed this concept of
non-locality to be applied to the mechanism of homoeopathy. This has also been
demonstrated in the works of Edward Whitmont, who
emphasises that the homoeopathic approach is finalistic and phenomenalistic,
rather than causalistic – thus a symbol,
representational of a whole. Bohm suggested that in
the implicate order, mind and matter can be looked at in a similar way, that
quantum mechanics may see mind and matter as enfolded. He has further stated
that within the framework of quantum mechanics, phenomenal reality comes about
from a deeper order in which it is enfolded or implied. In order to extrapolate
on the meaning of this
innate property of implication in the physical universe, he uses the
example of the hologram; each part of the photographic plate contains
information about the whole.
The whole is unfolding from each region on the photographic plate (Davies
& Brown 1986:118).
Wallach (2000) also suggests that the effect of the homoeopathic remedy
is not a causal one as would be explained in an orthodox sense, but rather
through a system of
“signs” or concepts. Thus a universal non-local and a causal means are
present within the substance. This universal interconnectedness of all creation
may be the mechanism whereby homoeopathy acts through consciousness.
The work of Carl Jung would underline this very strongly. The occurrence
of archetypal symbols and the universal meaning contained therein is very
appropriate in the scientific domain – from physics and psychology to
homoeopathy and philosophy (Jung 1993:384).
This was also a conclusion reached in a discussion between Jung and
Pauli; that psychological states and physical events could be a causally
connected through an element
of meaning (Davies 2001:38).
The homoeopathic remedy thus becomes a symbol with a specific element of
meaning. The meaning that is so contained in the remedy as symbol, may also
serve as a deeper understanding of the fundamental principle in homoeopathy: “Similia Similibus Curentur” – let like be cured by like. It is important to
also extend this concept to Anthroposophy, which is based ultimately, on a
foundation of “spiritual science” where the unseenis
in fact the template for physical matter and its behaviour.
Speaking on the nature of man, Rudolf Steiner had remarked that “(the
understanding of man) rests upon the recognition of a hidden something behind that
which is
manifest to the outer senses and to the intellect brought to bear upon
their perceptions.
These senses and this intellect can apprehend only a part of all that
(is) the total human entity...” (Wilson 1985:10).
Oppenheimers words , to some degree, are echoed
in Steiner’s insistence that total material fails to account for the
complexities of the universe and of human existence.
NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY
Nuclear magnetic resonance (NMR) spectroscopy is a characterization
technique whereby the physico-chemical
environment of specific nuclei is deduced from information obtained about the
nuclei within an analysed sample. This in turn
provides a means whereby the molecular identity of a substance may be
expressed and described in scientific terms.
NMR RESEARCH IN HOMOEOPATHY
In 1965 Barnard developed the idea of the potentization
process resulting in the transfer of informational content from the solute to
the solvent. He proposed the formation
of stereospecific solvent molecule polymers in
water (solvent). These polymers would be structurally determined by the
original solute and the spatial changes it caused in
the solvent and were theorised to be induced to self-replicate and split
by the energy provided by succussion (Sacks:1983).
Smith and Boericke investigated Barnard’s
theory by method of NMR spectroscopy. Their experiment with various Sulphur
potencies, lead to the following conclusions
with regard to the H2O and OH sections of the spectra: Solvent structure
is changed in unsuccussed serial solutions when
compared to undiluted solvents.
Solvent structure is further changed by succussion
of serial dilutions when compared to unsuccussed
dilutions and undiluted solvent.
These changes become more extreme as the dilutions approach and pass the
negative function of Avogadro’s number.
These results extended to the hypothesis that catalysis of exchange
between C2H5OH and H2O protons of ethanol-water mixtures is very likely to take
place in homoeopathic preparations (Weingartner:1990).
Additional proof of the effect of succussion,
compared to identical dilution, was carried out by studying the effect of
serial succussions and dilutions of bradykinin triacetate (BKTA). It was established, that
there is a definite, recurring, reproducible change in NMR patterns by succussion and that these patterns are reproducible and may
be caused by water polymers (Smith & Boericke:1968).
Sacks (1983) conducted an experiment on 32 different potencies of high
and ultra high dilutions; Sulphur in mother tincture, 6x, 12x, 22x, 23x, 24x,
25x, 30x, 200x, 1M, 10M, 50M, CM and seven other remedies
in 30C. It was graphically shown that, whereas the ethanol-water control
spectra did not vary throughout the experiment, the hydroxyl region changed in
a variety of ways.
Practically all of the succussed high dilution
samples were distinctly different from ethanol, and many were different from
each other. However, no clear pattern of peak shape was discernible from
Sulphur 6x through CM, nor among the seven remedies of 30C potency.
Different quantities of ethanol-water control caused very minor
variations in size, but no variation in shape of hydroxyl peaks. Different
quantities of succussed high dilutions distinctly
altered the shape of the curve or the hydroxyl peaks. Clear differences emerged
on the NMR spectra of succussed high dilutions and
ethanol-water control in this study. The differences lie in the hydroxyl
regions. In ethanol-water controls two peaks are visible, one belonging to
ethanol and the other to water. In the succussed
samples the peaks become less distinct and even merge. Sacks concluded that
this may indicate differences in relationship of ethanol and water hydroxyl
groups. It may also be some manner of artefact. In a statistical trial
conducted by Weingartner (1990), the 1H NMR spectra
of homoeopathic Sulphur-potencies and their solvent were compared with
each other. At a significance level of 99.9% the recorded spectra could
be distinguished with respect to the relative intensities of the H2O and the OH
signals.
It was hypothesised that the 1H NMR spectra of Sulphur D23 and its
solvent have statistically different relative intensities of the H2O and OH
signals with respect to
the mean intensity of the CH2 signal.
It was confirmed in three separate runs that the quantities in the 1H
NMR-spectra of Sulphur D23 differed systematically from those of ethanol 87%
spectra and that the
same quantities of Sulphur D13 do not. No distinction between the
Sulphur D23 and the ethanol 87% group of samples could be made in the shifts
relative to 1.186 ppm,
in the integrals and in the coupling constants. The intensities of the
H2O and the OH signals relative to CH2 however, were drastically different in
the Sulphur D23 and
in the ethanol group. Sulphur D13 spectra did not differ so obviously
from the solvent-spectra. In the experiment, different spectrometers were
utilized to exclude the possibility of machine artefact (Weingartner:1990).
Lasne et al. showed that in higher degrees of
dilution, significant differences exist in spin-spin relaxation times between potentized substances and their solvents.
This result was also obtained independently by a coworker
of Resh and Gutmann
(Weingartner:1990).
In 1992 Demangeat performed a study comparing vortexed potencies of Silicea (in
0.9% saline solution) and versus pure saline solvent prepared according to the
French Homoeopathic Pharmacopoeia. Dilution levels were measured at levels
ranging from 1.66 x 10-5 to 1.66x 10-29 mol/l. The relaxation times T1 as well
as the T1/T2 values
of the pure saline solution were found to be lower than the values of
the Silicea potencies. The differences were
statistically significant at p<0.034 and p<0.018,
respectively. Comparison was also made with agitated dilutions of
solvent (0.9 % NaCl), and distilled water. T1/T2
values were observed to increase with the addition
of a solute in all solvent comparisons.
Theories given for the observed differences were the interaction of solutes
with water molecules through hydrogen bonding, and perhaps also electrostatic
forces linked to
their dipolar moment. Isotope effects similar to the theories of Berezin (1991) were also considered, as was the structure
breaking effect of the solute first introduced by
Resch and Guttman (Davies
2001:30).
More recently, in comparing Hahnemannian and Korsakovian potentising methods
in analogous samples, Davies (2001) confirmed that statistically significant
differences was found in almost all intra-potency comparisons for each method
and its respective control. This confirmed a hypothesis that changes occur
during potentizations which are specific to each
method employed. He further draws the conclusion that the method of dilution is
an integral part of the remedy’s physico-chemical
information content,
and therefore not equivalent in homoeopathic practice.
SUMMARY
It is clear that the principle of dynamization
is of critical importance, both in establishing a medically active preparation,
as well as in establishing a particular physico-chemical
identity in a sample. The effect of succussion is
arguably investigable through NMR and creates the possibility of further
understanding of homoeopathy as a whole, in particular the potentization
process and its effects.
A comparison between Hahnemannian and Anthroposophical potentization
thus warrants investigation. Not only are Anthroposophical
remedies analogous to homoeopathic preparations, save for the succussion method, but they are also commonly utilized interchangeably,
especially in the OTC (= over-the-counter drug) industry. A comparative
analysis would thus be informative as to its actual similarity.
With various NMR studies done analysing homoeopathic preparations, Anthroposophical preparations have not been subjected to
the same scrutiny.
With the only variable in this experiment being the succussion
technique, any result, positive or negative, will reflect on the process of succussion and associated theories and allow greater
enquiry with regards to their appropriateness and accuracy.
CHAPTER THREE: MATERIALS AND METHODS
PRODUCTION OF SAMPLE POTENCIES
All preparation and investigation of samples was conducted within a
laboratorial setting. Thus the only variables that may successfully be controlled
are those within the laboratory. The researcher made allowance for
uncontrollable variables, for example cosmological influences, as purported to
be of significance in Anthroposophical thought.
All samples were prepared by hand under laminar flow at Pharma Natura laboratories,
according to standardized GHP, as per method 5a and Anthroposophical
Extended Medicine methodology respectively.
Serial dilution was maintained in 1:10 with 62%, 43%, 30% and 15%
ethanol successively up to the 4th level of dilution (D4).
Thereafter a concentration of 15% ethanol
was maintained in all samples. Succussion of
the Hahnemannian (homoeopathic) sample was constant
at 100 succussions per dilution level. Both anthroposophical samples underwent succussion
for a duration of two and a half minutes per dilution level. All potencies were
prepared from the same source of Pulsatilla pratensis Ř, in
glassware
of identical nature and batch. Care was also taken to ensure that all
ethanol used in deconcentration was of the same source.
A parallel process of production was employed to minimize differences in
the conditions surrounding the manufacture of the potencies. Production of all
samples was conducted by a singular individual, within the same time period
(one day).
On completion all preparations were stored in amber glass bottles to
reduce the possible influence of sunlight.
Completed samples were packaged in insulated containers to minimize
movement within the samples and to maintain a constant temperature during transportation.
Precautions were taken to eliminate the possibility of any contaminants
in the samples, which may influence NMR spectra readings. This was done
according to the standards in the British Pharmacopoeia for sterilisation of
glass apparatus. All glass bottles used in the preparation of samples were
rinsed in distilled water and underwent dry heat sterilization at 180° C for 35
minutes. All the bottles were left to cool before the manufacturing commenced.
NMR MEASUREMENT OF SAMPLES
NMR analysis of the samples was conducted at the University of Kwa-Zulu Natal, department of Chemistry.
The operation and running of the machine was performed by Mr. Craig
Grimmer, resident NMR technician at the university. The spectrometer used was a
Varian 500MHz INOVA Spectrometer, having a 5mm broadband switchable probe and a
5mm inverse detection probe. The pulse angle was set at 90° and the temperature
was thermostatically controlled, remaining constant at 25° C. 1.75 ml of each
sample was drawn into a coaxial tube by means of a micropipette. The experiment
was conducted using acetone as
an external lock and ethanol as the reference.
NMR-spectra were recorded of the CH3, CH2, H2O and OH signals of three
separate samples of each respective preparation and expressed in the chemical
shift and relative integration values. Readings were repeated sixteen times for
every sample to eliminate inconsistencies and inaccuracies.
Spectra of the OH, H2O, CH2, and CH3 signals were recorded and expressed
in terms of chemical shifts and relative integration values. All data were
transferred into Microsoft Word and printed.
STATISTICAL ANALYSIS
Chemical shift and relative integration values of CH2, CH3, OH and H2O
signals were recorded and utilized in statistical analysis. Chemical shift
values were then measured for reliability using Cronbach’s
alpha. Tests for normality were done by means of the Kolmogorov-Smirnov
and Shapiro-Wilk tests.
A significant outcome implies the conclusion that the distribution is
not normal, whereas a non-significant outcome implies only that no deviation
from normality has been demonstrated - not that it doesn’t exist. A negative
result however, is some evidence of normality and all the evidence we have,
which leads us to carry out the analysis as
if normality had been demonstrated.
The choice of the test to use depends on both the sample size and
whether the user would rather err on the side of being too conservative or the
opposite.
The two tests of normality used for this statistical analysis was the Kolmogorov-Smirnov test and the Shapiro-Wilk
test.
The Shapiro-Wilk test tends to reject the null
hypothesis more readily than one would wish; whereas the Kolmogorov-Smirnov
test is too conservative, retaining the null hypothesis too often.
CHAPTER FOUR: THE RESULTS
CRITERIA GOVERNING THE ADMISSIBILITY OF DATA
The sensitive nature of the experiment demands great care and precision
during every stage of the experiment. Care was taken to exclude external
factors inasmuch was possible that may affect the integrity of the samples. The
appropriate precautions were taken in manufacturing, storage, transport and
analysis. The effect of electromagnetic radiation on the samples is uncertain.
Great effort was made in the prevention of variability in samples and analysis,
as already explained in 3.1 and 3.2. Samples were constantly maintained under
the same conditions and atmospheric exposure was kept to a minimum.
Similarly, contamination was avoided by the singular use of glassware
throughout.
Data containing chemical shift and relative integration values of the
CH3, CH2, H2O and OH peaks were subjected to statistical analysis as set out in
3.3.
SUMMARY OF STATISTICAL ANALYSIS
From the above results it is noted that there are no statistically significant
differences between Wala, Weleda
or Hahnemannian methods of potentization
reflected in
the chemical shift values of the OH peaks.
There are, however, statistically significant differences in the
chemical shift values of both the CH2 and CH3 peaks between Wala,
Weleda and Hahnemannian
methods
of potentization, interchangeably.
In terms of the relative integration values, statistically significant
differences were noted between Wala and Hahnemannian methods of potentization
for both the OH and CH3 peaks. Wala and Weleda methods of potentization
revealed statistically significant differences only in the CH3 peaks of the
relative integration values.
DISCUSSION
Of all the components in the NMR study, the chemical shift is more
sensitive and indicative of any changes in the local environment than the
relative integration values.
Thus the results of this study can best be explained in considering the
chemical shift values, as it is most susceptible in reflecting structural
changes in the sample being investigated.
In the intergroup comparisons, CH2 and CH3 showed statistically
different results in terms of chemical shift. Due to the molecular structure of
ethanol, the polar hydroxyl group exerts an influence on the molecular electron
configuration of ethanol – most notably at the location of the methylene group.
The chemical shift peaks are also subject to local diamagnetic effects,
local paramagnetic effects and electric field effects. Anisotropic effects due
to neighbouring groups in
the molecule would generate local magnetic field effects, and these
would in turn affect chemical shifts. In the case of ethanol, the combination
of the local group anisotropy and the electric field effects due to the polar
hydroxyl group makes the CH2 chemical shift most susceptible to external
influences like dynamization. This is affirmed in
statistically significant differences found in the CH2 chemical shift values of
all three methods analyzed.
A theory on the influence of serial dilution and succussion
on the specific organization of molecules within homoeopathic microdilutions, as proposed by Anagnostatos
et al. (1991), is adopted as base wherefrom the findings of this project may
find support.
The clathrate-model states that through
grinding and dilution into a solvent, the formation of small clusters of an
original substance occurs in a solution.
These clusters exhibit substantial stability and possess the
characteristic and highly symmetrical shape of the specific substance.
Water molecules surrounding each cluster forms hydrogen bonds with one
another, resulting in a shell with a shape similar to that of the cluster.
These formations of water molecules are termed clathrates.
Subsequent succussion allows the clusters to
overcome the water cohesion forces and relocate to a new position, with
resultant new clathrates forming around it.
After the cluster relocation, the broken original clathrate
shell repairs itself, possessing a void in the interior, similar to the
relocated cluster. This now becomes a “core-clathrate.”
Another “mantle-clathrate” is now formed around the
core clathrate.
The role of substance clusters is now taken over by the core clathrates. Their symmetric and compact structure allows
for extra stability and, thus, behaviour like large
complex molecules, i.e., molecules with a much larger mass and different
inertial properties than regular water molecules.
With serial dilution and succussion this
process is perpetuated. It is thus put forward that the properties of the
initial substance can be traced by the properties of the
shaped voids in the solvent. Thus the specific homoeopathic remedy,
resulting from serial dilution and dynamization may
have the characteristic properties of an original substance not physically
present.
The results of this study can be interpreted on the basis of the clathrate model.
The potentization methods in all three methods
analyzed serve to dynamically redistribute the energy within higher order
structures.
Hahnemannian potentization
provides a linear, additive effect on the remedy solution, resulting in the
effects as described by the standardized clathrate
model.
The Wala method applies a non-linear energy
distribution to clathrate structures, thus resulting
in, and re-enforcing the formation of higher order clathrate
structures.
In order to explain differences in the results associated with the Weleda method one would need to take a quantum mechanical
approach. According to Heisenberg’s Uncertainty Principle, the clathrates are non-static, which means they possess an
inherent natural frequency associated with them. The Weleda
method employs a harmonic distribution of energy at a fixed
frequency. This may result in resonance between the input energy and the
clathrate oscillations. In turn, this may serve to
break up the clathrates and serve to create higher
order
structures within the remedy, via a dynamic re-distribution of energy.
It may thus be concluded that the effect of vorticity
(Wala), non-linearity (Weleda)
and linearity (Hahnemannian) in the applied force of dynamization, all serve to alter the remedy to
varying extents.
CONCLUSIONS
The results of this study showed that statistically significant
differences were observed in the chemical shift values of the CH2 and CH3
signals for all three methods investigated.
Relative integration values showed significant differences between the Wala and Weleda method for the
CH3 signal, and between the Wala and Hahnemannian method
for both OH and CH3 signals.
Based on the results of this investigation, it may thus be reasoned that
the method of dynamization does play a significant
and crucial role in establishing a particular
molecular environment within a succussed
dilution, and therefore, in the development of a distinct physico-chemical
identity of a remedy. It may therefore also follow
that the hypothesis that different methods of dynamization exert an
individualizing effect on a remedy and thus may imply inherent differences, is
satisfied.
RECOMMENDATIONS
1. Standardization of the pharmaceutical process Strict standardisation
of the potentization process is necessary to allow
scientific evaluation and reproducibility. As potential human error is
unavoidable in any procedure, succussion may
therefore require control with calibrated machinery.
2. Magnetic field strength
The effect of the strong magnetic field used in high resolution NMR
spectroscopy may have an uncertain effect on the samples analysed. The
utilization of spectrometers of different field strengths, ranging from very
low to very high may be advisable in analysing ultra high dilution samples.
3. External factors
Although external factors in this study are controlled as far as
possible, concerns may still exist regarding the exact nature of the potentising process, particularly in the production of Anthroposophical samples. Within Anthroposophy, certain
prohibitions are placed on the production process, e.g. potentising
may only be carried out between 2.30 h. and 10.30 h. and between 14.30 h. and
22 h. Also, when potentising metals, the day of the
month must be indicated as suitable according to the Wala
and Weleda Potentising
Calendars; potentising is prohibited during certain
celestial occurrences, e.g. sun and moon eclipses.
The extent of these theoretical influences can not easily be verified
scientifically. Still, strict observations of these parameters establish
guidelines in standardizing samples.
Chemical influences, e.g. the absorption of moisture during the potentising process and variations in factors like at
atmospheric oxygen may play a role, one may also question
excessive control as unnatural to the potentization
process. One must therefore attempt to replicate the potentization
process as is done in practise as closely as possible, with
as few variations as possible (Davies 2001).
4. Analysis of a wider variety of substances and samples
The use of a wide variety of substances prepared according to
standardised methods would allow a better comparison of trends that arise
within NMR spectroscopic analysis.
Large amounts of anomalous results instead of definite trends could
indicate that the use of NMR spectroscopy should be re-evaluated as a tool for
analysing homoeopathic substances.
Vorwort/Suchen Zeichen/Abkürzungen Impressum