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 4^th 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 Ø, 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