This is part of the Homeopathy journal club described here:

www.badscience.net/?p=490

doi:10.1016/j.homp.2007.03.006    
Copyright © 2007 Elsevier Ltd All rights reserved. The history of the Memory of Water

Yolène Thomas^{, a,
a}Institut Andre Lwoff IFR89, 7, rue Guy Moquet-BP8, 94 801 Villejuif Cedex, France
Received 26 March 2007;  accepted 27 March 2007.  Available online 31 July 2007.

‘Homeopathic dilutions’ and ‘Memory of Water’ are two expressions capable of turning a peaceful and intelligent person into a violently irrational one,’ as Michel Schiff points out in the introduction of his book ‘The Memory of Water’. The idea of the memory of water arose in the laboratory of Jacques Benveniste in the late 1980s and 20 years later the debate is still ongoing even though an increasing number of scientists report they have confirmed the basic results.

This paper, first provides a brief historical overview of the context of the high dilution experiments then moves on to digital biology. One working hypothesis was that molecules can communicate with each other, exchanging information without being in physical contact and that at least some biological functions can be mimicked by certain energetic modes characteristics of a given molecule. These considerations informed exploratory research which led to the speculation that biological signaling might be transmissible by electromagnetic means. Around 1991, the transfer of specific molecular signals to sensitive biological systems was achieved using an amplifier and electromagnetic coils. In 1995, a more sophisticated procedure was established to record, digitize and replay these signals using a multimedia computer. From a physical and chemical perspective, these experiments pose a riddle, since it is not clear what mechanism can sustain such ‘water memory’ of the exposure to molecular signals. From a biological perspective, the puzzle is what nature of imprinted effect (water structure) can impact biological function. Also, the far-reaching implications of these observations require numerous and repeated experimental tests to rule out overlooked artifacts. Perhaps more important is to have the experiments repeated by other groups and with other models to explore the generality of the effect. In conclusion, we will present some of this emerging independent experimental work.

Keywords: high dilution; memory; water; molecular signal; audio-frequency oscillator; computer-recorded signals

Article Outline

Historical overview: the early history of high dilution experiments

Exploring the physical nature of the biological signal

From high dilution to digital biology

The present situation

Acknowledgements

References

Historical overview: the early history of high dilution experiments

Presenting a brief history of what is known as the ‘Memory of Water’ is not an easy task mainly because one of the main actors, Jacques Benveniste, is no longer with us (Figure 1). There are always many controversies around cutting edge science, and especially with those whose lives have been spent pursuing unorthodox trails.

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Figure 1. Jacques Benveniste 1935–2004.

I first met Benveniste during a FASEB meeting in Atlanta in 1981 and joined his laboratory a few years later to set up my own Immunology team. I had the good fortune of being able to collaborate with him for over 16 years. At that time, he was at the top of his fame and gained an international reputation as a specialist on the mechanisms of allergies and inflammation with his discovery of the ‘Platelet Activating Factor’ (paf-acether) in 1970.^{[1] and [2]} Throughout his long career, working both in the US and in France, he was responsible for the development of new ways of approaching inflammation including the patenting by the French National Institute of Health and Medical Research (INSERM) of his innovative allergy test using blood cells called basophils (FR-patent-7,520,273). Jacques’ research into allergies took him deep into the mechanisms which create such responses: understanding how the smallest amount of a substance affects the organism. The life and work of Jacques Benveniste was not only written in water.

In the early 1980s, while heading up the unit INSERM 200, Jacques took a new member onto his staff, a young medical doctor, Bernard Poitevin, whose side-interest was homeopathy. ‘He asked me if he could try my basophil degranulation test on some homeopathic preparations’, Jacques recalled, ‘and I remember distinctly saying “OK, but all you will be testing is water”.’ Thus, Jacques expressed his skepticism but accepted the proposal.

After 5 years of research they empirically observed that highly dilute (i.e., in the absence of any physical molecule) biological agents nevertheless triggered the relevant biological systems. Intrigued but cautious, Jacques was a man who adhered to the facts. He ordered a two-year long series of retests, but the same results kept recurring. Finally, Poitevin and Benveniste submitted two papers which were published in peer review journals.^{[3] and [4]} Here, the work was treated as conventional research like many other manuscripts from peer-reviewed journals which can be found in the scientific literature on the effect of high dilutions (HD) (review in^{[5] and [6]}).

Following accepted scientific practice, Jacques then asked other laboratories to try to replicate the findings. In 1988, scientists from six laboratories in four countries (France, Canada, Israel and Italy) co-authored an article showing that highly diluted antibodies could cause basophil degranulation. This was established under stringent experimental conditions such as blind double-coded procedures. Further, the experimental dilution (anti-IgE) and the control one (anti-IgG) were prepared in exactly the same manner, with the same number of dilution and agitation sequences. The article was submitted to Nature.⁷ Nature’s referees could not fault Benveniste’s experimental procedures but could not comprehend his results. How can a biological system respond to an antigen when no molecules of it can be detected in the solution? It goes against the accepted ‘lock-and-key’ principle, which states that molecules must be in contact and structurally match before information can be exchanged. In the paper, Jacques suggested that specific information must have been transmitted during the dilution/shaking process via some molecular organization occurring in the water.

Finally, the editor of the journal, John Maddox agreed to publication, on condition that a ‘committee’ could verify Benveniste’s laboratory procedures. In July 1988, after two weeks after publication, instead of sending a committee of scientific experts, Maddox recruited—James Randi, a magician, and Walter Stewart, a fraud investigator. The three of them spent 5 days in the laboratory. Well, you all know what followed. Nature’s attempted debunking exercise failed to find any evidence of fraud. Nevertheless, they concluded that Benveniste had failed to replicate his original study.⁸ This marked the beginning of the ‘Water Memory’ war, which placed him in a realm of ‘scientific heresy’. As Michel Schiff later remarked in his book: ‘INSERM scientists had performed 200 experiments (including some fifty blind experiments) before being challenged by the fraud squad. The failure to reproduce⁸ only concerned two negative experiments’.⁹ Benveniste replied to Nature¹⁰ and reacted with anger, ‘not to the fact that an inquiry had been carried out, for I had been willing that this be done… but to the way in which it had been conducted and to the implication that my team’s honesty and scientific competence were questioned. The only way definitely to establish conflicting results is to reproduce them. It may be that we are all wrong in good faith. This is not crime but science…’.

As a consequence of the controversy that ensued, Jacques became increasingly isolated. Nonetheless the team repeated the work on a larger scale, entirely designed and run under the close scrutiny of independent statistical experts, and confirmed the initial findings in Nature.¹¹ These further experiments have been coolly received or ignored by most scientists at least partly because, given Jacques’ now-acrimonious relationship with Nature, they were published in a less renowned journal.

To date, since the Nature publication in 1988, several laboratories have attempted to repeat Benveniste’s original basophil experiments. Most importantly, a consortium of four independent research laboratories in France, Italy, Belgium, and Holland, led by M. Roberfroid at Belgium’s Catholic University of Louvain in Brussels, confirmed that HD of histamine modulate basophil activity. An independent statistician analyzed the resulting data. Histamine solutions and controls were prepared independently in three different laboratories. Basophil activation was assessed by flow-cytometric measurement of CD63 expression (expressed on cytoplasmic granules and on the external membrane after activation). All experiments were randomized and carried out under blind conditions. Not much room, therefore, for fraud or wishful thinking. Three of the four labs involved in the trial reported statistically significant inhibition of the basophil degranulation reaction by HD of histamine as compared to the controls. The fourth lab gave a result that was almost significant. Thus, the total result over all four labs was positive for histamine HD solutions.^{[12] and [13]} ‘We are,’ the authors say in their paper, ‘unable to explain our findings and are reporting them to encourage others to investigate this phenomenon’.

Different attempts have been made to substantiate the claim that serial dilution procedures are associated with changes in the water’s physical properties (^{[14] and [15]}and see Louis Rey contribution in this issue pages 170–174). Yet, the challenge of understanding the mechanisms of how HDs work, and the role of water in them, is a difficult one to say the least. Several possible scenarios have been suggested. One proposed by Giuliano Preparata and Emilio Del Giudice, is that long range coherent domains between water molecules (quantum electrodynamics, QED) gives high dilution laser-like properties.^{[16] and [17]} When the field matches the kinetic of the reaction, the latter becomes functional as the optimal field strength as for a radio receiver. It was to a scientific meeting in Bermuda that took place a few months before the Nature ‘affair’ erupted that these two physicists working at Milan University brought the theoretical basis for the memory of water. Another scenario predicts changes in the water structure by forming more or less permanent clusters.¹⁸ Other hypotheses will be discussed in this issue. High dilution experiments and memory water theory may be related, and may provide an explanation for the observed phenomena. As M. Schiff points out, only time and further research will tell, provided that one gives the phenomena a chance.⁹

Exploring the physical nature of the biological signal

Despite the difficulties after the Nature fracas, Jacques and his now-depleted research team continued to investigate the nature of the biological activity in high dilutions and aimed at understanding the physical nature of the biological signal. In his Nature paper, Jacques reasoned that the effect of dilution and agitation pointed to transmission of biological information via some molecular organization going on in the water. The importance of agitation in the transmission of information was explored by pipetting dilutions up and down ten times and comparing with the usual 10-s vortexing. Although the two processes resulted in the same dilution, basophil degranulation did not occur at HD after pipetting. So transmission of the information depended on vigorous agitation, possibly inducing a submolecular organization of water or closely related liquids (ethanol and propanol could also support the phenomenon). In contrast, dilutions in dimethylsulphoxide did not transmit the information from one dilution to the other. In addition, heating, freeze-thawing or ultrasonication suppressed the activity of highly diluted solutions, but not the activity of several active compounds at high concentrations. A striking feature was that molecules reacted to heat according to their distinctive heat sensitivity, whereas all highly diluted solutions ceased to be active between 70 and 80 °C. This result suggested a common mechanism operating in HDs, independent of the nature of the original molecule. In addition, in 1991 and in collaboration with an external team of physicists (Lab. Magnetisme C.N.R.S.-Meudon Bellevue, France), it was shown in twenty four blind experiments that the activity of highly dilute agonists was abolished by exposure to a magnetic field (50 Hz, 15×10⁻³ T, 15 min) which had no comparable effect on the genuine molecules. Moreover, it is worth pointing out that a growing number of observations suggest the susceptibility of biological systems or water to electric and low-frequency electromagnetic fields.^{[19], [20] and [21]} In addition, what is suggested from the literature is a possible role of electromagnetic fields regarding informational process in cell communication.^{[22], [23] and [24]}

At this stage, Jacques hypothesized that transmission of this ordering principle was electromagnetic in nature and move on to the idea that molecules could communicate via specific electromagnetic waves. If so, what molecule vibration modes are efficient and how can these signals be used to mimic some of the biological functions of a molecule without its physical presence?

From high dilution to digital biology

It was at the beginning of the nineties that a homeopathic physician, E. Attias convinced Jacques to try out an electrical device that he claimed transmitted chemical information. After a few positive trials with this machine, Jacques had another one built, which was used for later experiments. This second device was essentially a standard audio amplifier that, when connected to another coil, behaves as an audio-frequency oscillator. Between 1992 and 1996, we performed a number of experiments showing that we could transfer, in real time, molecular signals indirectly to water or directly to cells. Briefly, cells were placed in a 37 °C humidified incubator on one coil attached to the oscillator, while an agonist (or vehicle as control) was placed on another coil at room temperature. Here, the transfer was not a two step-process, as when water acts as an intermediary recipient of the molecular signal. In one such exploration, we showed that molecular signals associated with a common phorbol ester (phorbol-myristate-acetate) could be transmitted by physical means directly to human neutrophils to modulate reactive oxygen metabolite production. In 1996, I submitted an article about these experiments to several prestigious journals. The article was flatly rejected each time, on the grounds that we could not explain the underlying mechanism, in spite of the referees’ general opinions that our work was ‘state-of-the-art’ and was ‘provocative and intriguing and we have gone to great lengths to try to eliminate any biological variables that could bias our results.’ It was finally published in 2000.²⁵ Appended to this article were two affidavits, one from a French laboratory (F. Russo Marie, INSERM U332, Paris, France) testifying that they supervised and blinded the experiments we did in this laboratory; the other from an US laboratory (W. Hsueh, Department of Pathology, Northwestern University, Chicago) testifying that they did some preliminary experiments similar to ours, without any physical participation on our part, and detected the same effect.

Because of the material properties of the oscillator and the limitations of the equipment used, it is most likely that the signal is carried by frequencies in the low kilohertz range.²⁶ These considerations led to the establishment in 1995 of a more sophisticated procedure for the recording and retransmission of the molecular signals. DigiBio, a company that Jacques had set up in 1997 to finance his research, obtained in 2003 an approval for one of his French patents by the US Patent Office (6,541,978: method, system and device for producing signals from a substance biological and/or chemical activity). The characteristics of the equipment are described in Figure 2 and in.²⁶ Briefly, the process is to first capture the electromagnetic signal from a biologically active solution using a transducer and a computer with a sound card. The digital signals are stored (Microsoft sound files *.wav). The signal is then amplified and ‘played back’, usually for 10 min, from the computer sound card to cells or organs placed within a conventional solenoid coil. The digitally recorded signals can also be played back into untreated water, which thereafter will act as if the actual substance was physically present.

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Figure 2. Schematic drawing of the computer-recorded signals: capture, storage and replay:

• Shielded cylindrical chamber: Composed of three superposed layers: copper, soft iron, permalloy, made from sheets 1 mm thick. The chamber has an internal diameter of 65 mm, and a height of 100 mm. A shielded lid closes the chamber.

• Transducers: Coil of copper wire, impedance 300 Ω, internal diameter 6 mm, external diameter 16 mm, length 6 mm, usually used for telephone receivers.

• Multimedia computer (Windows OS) equipped with a sound card (5–44 KHz in linear steps), (Sound Blaster AWE 64, CREATIVE LABS).

• HiFi amplifier 2×100 watts with an ‘in’ socket, an ‘out’ socket to the speakers, a power switch and a potentiometer. Pass band from 10 Hz to 20 kHz, gain 1–10, input sensitivity +/− V.

• Solenoid coil: Conventionally wound copper wire coil with the following characteristics: internal diameter 50 mm, length 80 mm, R=3.6 Ω, three layers of 112 tums of copper wire, field on the axis to the centre 44×10⁻⁴ T/A, and on the edge 25×10⁻⁴ T/A.

All links consist of shielded cable. All the apparatus is earthed.

From 1995 to the present, several biologically active molecules (eg histamine, acetylcholine, caffeine, PMA, Melagatran… even homeopathic medicines such as Arnica montana) have been recorded, digitized and replayed to biological systems sensitive to the original molecular substance. Several biological models were used. The first one was a commonly used system by pharmacologists, called the Langendorff preparation. By injecting different vasoactive substances into the coronary artery of an isolated, perfused guinea pig heart and measuring the coronary flow, you can quantify the vasoconstricting or vasodilating effect of the agent. In typical experiments, the signal of acetylcholine (or water as control), a classical vasodilating molecule was recorded and digitized. The signal was then amplified and ‘played’ back onto water. The signal-carrying water is then injected into the isolated heart, and consequently the coronary flow increased. Interestingly, atropine, an acetylcholine inhibitor, inhibited both the effects of the molecular acetylcholine as well as the digital signal of acetylcholine. Of note, the order of the conditions and their repetitions was always randomized and blinded. Other models include: human neutrophil activation; detection of the recorded signal of bacteria (E. Coli and Streptococcus) by playing them to a biological system specific to the bacterial signal and; the inhibition of fibrinogen coagulation by a Direct Thrombin Inhibitor. Further details of three of these salient biological models have been previously described.²⁶ Together, these results suggested that at least some biologically active molecules emit signals in the form of electromagnetic radiation at a frequency of less than 44 kHz that can be recorded, digitized and replayed directly to cells or to water, in a manner that seems specific to the source molecules.²⁶

Assuming that we give credence to the phenomena described, one question naturally springs to mind: what do molecule vibration modes sound like? Can measurable signals been identified in the form of low frequency spectral components? Didier Guillonnet, an engineer in computer science, and at the time, a close collaborator of Jacques Benveniste admitted, ‘When we record a molecule such as caffeine, for example, we should get a spectrum, but it seems more like noise. We are only recording and replaying; at the moment we cannot recognize a pattern although the biological systems do.’ Jacques called this matching of broadcast with reception ‘co-resonance,’ and said it works like a radio set.

Among the various theoretical problems associated with such a signal, two appear particularly relevant. First, how is such information using water as an intermediary detected amongst much electromagnetic noise? In fact, it has been suggested that stochastic resonance is an important mechanism by which very weak signals can be amplified and emerge from random noise.²⁷ Second, the limitations of the equipment used here, suggest that the signal is carried by frequencies in the low kilohertz range, many orders of magnitude below those generally associated with molecular spectra (located in the infrared range). However, molecules may also produce much lower ‘beat’ frequencies (Hz to kHz) specific for every different molecule. The ‘beat frequency’ phenomenon may explain this discrepancy, since a detector, for instance a receptor, will ‘see’ the sum of the components of a given complex wave.²⁸ Clearly, more experimental and theoretical work is needed in order to unveil the physical basis of the transfer (and storage?) of specific biological information either between interacting molecules or via an electronic device.

Replicability: Although since the very beginning we have placed a great deal of emphasis on carrying out our work under the highest standards of methodology and that great effort has been made to isolate it from environmental artifacts, attempts to replicate these data in other laboratories yielded mixed results. For instance, in 1999, Brian Josephson, Nobel Laureate for Physics in 1973 invited Benveniste to the Cavendish Laboratory in Cambridge. He said, ‘We invited him to learn more about the research which seems both scientifically interesting and potentially of considerable practical importance. Jacques definitely recognized there was a problem with reproducing the effect. The situation seemed to be that in some circumstances you had reproduction and in others you didn’t; but the overall results were highly significant.’ We then realized the difficulty in ‘exporting’ a method, which is very far from conventional biology. There are many key variables that might be involved like, water purification, the container shape and material being used, the purity of chemicals, atmospheric conditions…. Only if these underlying variables are known could the experiments be reproducible. When the transfer is a two-step process using water as an intermediary support for transmitted molecular signals, it takes even more stringent conditions for the experiments to be repeatable. The digital signal is replayed onto the water, which may take or not take the signal depending, for instance, upon the local electromagnetic conditions. In this regard, it is interesting to note that the ‘informed water’ as in the HD experiments, loses its activity after heating or being exposed to magnetic fields.

More surprising and mysterious was the fact that in some cases certain individuals (not claiming special talents) consistently get digital effects and other individuals get no effects or perhaps block those effects (particularly when handling a tube containing informed water). The inhibition of fibrinogen–thrombin coagulation by a digitized thrombin inhibitor is a model particularly sensitive to experimenter effects and therefore may account for the difficulty in consistently replicating this experimental system. Despite the precautions taken to shield the information transfer equipment from magnetic or electromagnetic pollution, very little concern has been given to possible subtle human operator effects.²⁹ We dealt with this problem in some of our own studies and also in the course of one independent replication.³⁰

The present situation

Now that Jacques Benveniste is no longer with us, the future of the ‘digital biology’ is in the hands of those who have been convinced of the reality of the basic phenomena. It is up to them to explore with other models the generality of the effect. Most likely they will succeed if they combine full biological and physical skills to understand the nature of the biological signals.

In this regard, since June 2005, Luc Montagnier, the co-discoverer of HIV, is conducting experiments (detection of the recorded signals of various micro-organisms derived from human pathologies) which, confirm and extend the original finding. In 2006, he set up a company called Nanectis. Perhaps the most impressive emerging data is from a US group located in La Jolla, CA.

In barely four years, they have conducted novel research programs and expanded the original technology into a series of potential industrial applications. Since 2004, they have obtained several US patents (6,724,188; 6,952,652; 6,995,558; 7,081,747) and applied for International Patents (WO 06/015038: system and method for collecting, storing, processing, transmitting and presenting very low amplitude signals; WO 06/073491: system and method for producing chemical or biochemical signals). They can improve the molecular signal recording in particular by using both magnetic and electromagnetic shielding coupled to a superconducting quantum interference device (SQUID). The system records a time-series signal for a compound; the wave form is processed and optimized (selected noise amplitude, power setting…) to identify low-frequency peaks that are characteristic of the molecule being interrogated (Molecular Data Interrogation System, MIDS). The optimized signal is played back for various periods of time to sensitive biological systems. For instance, they describe one interesting model particularly relevant to the specificity of the molecular signal transmission effect. The arabinose-inducible bacterial system with a lac operon is inducible by signals from the L (+) arabinose form but not from the D (−) arabinose inactive isomer or the white noise control. Other systems include digital herbicides and plant growth regulator as well as pharmaceutical compounds such as Taxol ^®, a prototype for a class of anticancer drugs. For instance, in a classic in vivo mouse xenograft model, the digital Taxol was assessed by the growth inhibitory potential of a human breast tumor. The results revealed that tumor growth, by day 36, was as statistically significantly inhibited in the group treated with the Taxol signal, as it was in the control group treated with actual molecular Taxol. If these new experimental observations can be validated, we will have added yet another valuable piece to the puzzle.

Although a theoretical explanation of how the memory of water might work must still be explored, the fact that the effective transmission of molecular signals has now been observed by independent teams using different biological systems, provides a strong additional basis to suggest that the phenomena observed by Jacques were not due simply to laboratory artefacts.

Whatever knowledge ongoing and future investigation may bring, the difficult road that Jacques travelled by opposing the automatic acceptance of received ideas, will have contributed to sustaining freedom in scientific research and putting the emphasis back where it belongs, on observable fact.

Acknowledgments

I am grateful to Drs. Isaac Behar and Anita K. Gold for critical comments on the manuscript.

References

1 J. Benveniste, P.M. Henson and C.G. Cochrane, Leukocyte-dependent histamine release from rabbit platelets. The role of IgE, basophils, and a platelet-activating factor, J Exp Med 136 (1972), pp. 1356–1377. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

2 J. Benveniste, Platelet-activating factor, a new mediator of anaphylaxis and immune complex deposition from rabbit and human basophils, Nature 249 (1974), pp. 581–582. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

3 E. Davenas, B. Poitevin and J. Benveniste, Effect of mouse peritoneal macrophages of orally administered very high dilutions of silica, Eur J Pharmacol 135 (1987), pp. 313–319. Abstract | Abstract + References | PDF (543 K) | View Record in Scopus | Cited By in Scopus

4 B. Poitevin, E. Davenas and J. Benveniste, In vitro immunological degranulation of human basophils is modulated by lung histamine and Apis mellifica, Br J Clin Pharmacol 25 (1988), pp. 439–444. View Record in Scopus | Cited By in Scopus

5 H. Walach, W.B. Jonas, J. Ives, R. van Wijk and O. Weingartner, Research on homeopathy: state of the art, J Altern Complement Med 11 (2005), pp. 813–829. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

6 P. Bellavite, R. Ortolani, F. Pontarollo, V. Piasere, G. Benato and A. Conforti, Immunology and Homeopathy, Evidence-based Complementary Alternative Med 2 (2005), pp. 441–452. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

7 E. Davenas, F. Beauvais and J. Amara et al., Human basophil degranulation triggered by very dilute antiserum against IgE, Nature 333 (1988), pp. 816–818. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

8 J. Maddox, J. Randi and W.W. Stewart, High-dilution’experiments a delusion, Nature 334 (1988), pp. 287–290.

9 Schiff M. The Memory of Water. UK: Ed. Thorsons, 1995.

10 J. Benveniste, Dr Jacques Benveniste replies, Nature 334 (1988), p. 291. Full Text via CrossRef

11 J. Benveniste, E. Davenas, B. Ducot, B. Cornillet, B. Poitevin and A. Spira, L’agitation de solutions hautement diluées n’induit pas d’activité biologique spécifique, CR Acad Sci Paris 312 (1991), pp. 461–466.

12 P. Belon, J. Cumps and M. Ennis et al., Inhibition of human basophil degranulation by successive histamine dilutions: results of a European multi-centre trial, Inflamm Res (Suppl 1) 48 (1999), pp. S17–S18. View Record in Scopus | Cited By in Scopus

13 P. Belon, J. Cumps and M. Ennis et al., Histamine dilutions modulate basophil activation, Inflamm Res 53 (2004), pp. 181–188. View Record in Scopus | Cited By in Scopus

14 Lobyshev VI, Tomkevitch MS. Luminescence study of homeopathic remedies. In: Priezzhev AV, Cote GL (eds). Optical Diagnostics and Sensing of Biological Fluids and Glucose and Cholesterol Monitoring, Proceedings of the SPIE, Vol 4263. MAIK “Navka/Interperiodica” (Russia), 2001, pp 1605–7422.

15 V. Elia, S. Baiano, I. Duro, E. Napoli, M. Niccoli and L. Nonatelli, Permanent physico-chemical properties of extremely diluted aqueous solutions of homeopathic medicines, Homeopathy 93 (2004), pp. 144–150. SummaryPlus | Full Text + Links | PDF (154 K) | View Record in Scopus | Cited By in Scopus

16 E. Del Giudice, G. Preparata and G. Vitiello, Water as a free electric dipole laser, Phys Rev Lett 61 (1988), pp. 1085–1088. Full Text via CrossRef

17 G. Preparata, QED Coherence in Matter, World Scientific, Singapore (1995).

18 E.E. Fesenko and A.Y. Gluvstein, Changes in the state of water, induced by radiofrequency electromagnetic fields, FEBS Lett 367 (1995), pp. 53–55. Abstract | Abstract + References | PDF (294 K) | View Record in Scopus | Cited By in Scopus

19 R. Goodman and M. Blank, Initial interactions in electromagnetic field-induced biosynthesis, J Cell Physiol 199 (2004), pp. 359–363.

20 E. Ben Jacob, Y. Aharonov and Y. Shapira, Bacteria harnessing complexity, Biofilms (2004), pp. 239–263.

21 P.h. Vallée, J. Lafait, P. Mentré, M.O. Monod and Y. Thomas, Effects of pulsed low frequency electromagnetic fields on water using photoluminescence spectroscopy: role of bubble/water interface?, J Chem Phys 122 (2005), pp. 114513–114521. Full Text via CrossRef

22 G. Albrecht-Buehler, Rudimentary form of cellular ‘vision’, Proc Natl Acad Sci USA 89 (1992), pp. 8288–8292. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

23 M.W. Trushin, Studies on distant regulation of bacterial growth and light emission, Microbiology 149 (2003), pp. 363–368. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

24 B.W. Ninham and M. Boström, Building bridges between the physical and biological sciences, Cell Mol Biol 51 (2005), pp. 803–813. View Record in Scopus | Cited By in Scopus

25 Y. Thomas, M. Schiff, L. Belkadi, P. Jurgens, L. Kahhak and J. Benveniste, Activation of human neutrophils by electronically transmitted phorbol-myristate acetate, Med Hypotheses 54 (2000), pp. 33–39. Abstract | Abstract + References | PDF (188 K) | View Record in Scopus | Cited By in Scopus

26 Y. Thomas, L. Kahhak and J. Aissa, The physical nature of the biological signal, a puzzling phenomenon: the critical role of Jacques Benveniste. In: G.H. Pollack, I.L. Cameron and D.N. Wheatley, Editors, Water and the Cell, Springer, Dordrecht (2006), pp. 325–340.

27 K. Wiesenfeld and F. Moss, Stochastic resonance and the benefits of noise: from ice ages to crayfish and SQUIDS, Nature 373 (1995), pp. 33–36. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

28 C.N. Banwellk, Fundamentals of Molecular Spectroscopy, McGraw-Hill Publ., UK (1983) pp 26–28.

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Correspondence: Yolène Thomas, Institut Andre Lwoff IFR89, 7, rue Guy Moquet-BP8, 94 801 Villejuif Cedex, France. Tel.: +33(0) 1 49 58 34 81.

Homeopathy
Volume 96, Issue 3, July 2007, Pages 151-157
The Memory of Water

This is part of the Homeopathy journal club described here:

www.badscience.net/?p=490

doi:10.1016/j.homp.2007.05.001    
Copyright © 2007 Elsevier Ltd All rights reserved. Can water possibly have a memory? A sceptical view

José Teixeira^{, a,
a}Laboratoire Léon Brillouin (CEA/CNRS), CEA Saclay, 91191 Gif-sur-Yvette Cedex, France
Received 1 May 2007.  Available online 31 July 2007.

Homeopathic medicines are currently used in medical practice, despite controversy about their effectiveness. The preparation method is based on extremely high dilutions of many substances in water, far beyond any detectable level. For this reason, it has been suggested that water could retain a ‘memory’ of substances that have been dissolved in it before the successive dilutions. The paper stresses the fact that this idea is not compatible with our knowledge of pure water. If an explanation on physical grounds is to be found, research must focus in other aspects of the preparation, such as the presence of other molecules and dissolved gases.

Keywords: water structure; water dynamics; aggregation; metastability

Article Outline

Introduction

Pure water and homeopathic drugs

Properties of liquid water

Aqueous solutions

Conclusion

References

Introduction

Homeopathy and homeopathic medicines are widespread and well accepted by many doctors, pharmacists and patients. It is officially recognised by health authorities and agencies authorities and at a political level in many parts of the world. However, they are also criticized and attacked by others. It is not my purpose to participate actively in a complex debate that includes not only scientific aspects but also sociological and economic components. My contribution will address only the arguments relying on the properties of water and only from the physical view. Consequently, at best, it is a physicist’s view of the role played by water in homeopathic solutions.

To clarify this statement, I think that it is useful to remember that medicine is not only a science but also an art. A good doctor takes into account not only the sickness itself but also the patient, his environment and his psychological aspects. As a consequence, the prescription of a medicine fortunately includes a large part of empiricism. The goal is to restore a ‘normal’ state. One must admit that the complete knowledge of all the parameters intervening in a real situation is totally illusory and that this situation is unlikely to change in the foreseeable future. Anyway, even when the active principles and biological receptors are well known and identified, the reactions of different patients are not the same. To circumvent these inherent difficulties the performance of drugs is established via statistical analysis of large numbers of cases with a randomised double-blind methodology which implicitly recognizes the hidden role of components which escape to the normal scientific analysis of ‘exact sciences’.

Modern pharmacological research is based on a detailed knowledge of physical and chemical interactions between drugs and living cells. At the confluence of Biophysics and Chemistry, a more detailed and precise picture of those interactions is steadily emerging. Still, many traditional medications and frequently-prescribed drugs are currently used without such detailed knowledge of their action. For them, it is either difficult or useless to define the exact ‘paths’ from medicine to biology, then to chemistry and physics.

Pure water and homeopathic drugs

Many traditional drugs, as for example those extracted from plants, are extensively used in medicine. In some cases one or more active principles have been identified but even in such cases the exact action is usually not well understood at the level of chemical reactions or physical interactions taking place within living organisms. This situation is very common but has never been a limitation to prescribing drugs that have shown their effectiveness through many years of practical use. Certainly, in other cases, the interactions are known in great detail leading to the synthesis of well-defined drugs with specific and well controlled applications. But we remain far from a comprehensive and detailed knowledge of the action of drugs on living organisms.

Homeopathic drugs fall, at least partially, into the first category. Their use has been validated by real or supposed successes, the frontier of the two being probably irrelevant from the point of view of the patient. But there is an essential difference between traditional or ‘natural’ medicine and homeopathy. The latter is much more recent and based in a quasi philosophical concept (similia similibus curentur) stated by Hahnemann, perhaps by analogy with the contemporary first studies of immunization. With modern science, it should, in due course, be possible to understand the mechanisms of action of natural substances and of homeopathic drugs. For natural substances the search for the active principles has been successful in some cases; in others, it has been simply assumed that they are present but the level of interest of the drug or available resources has not justified further studies.

With homeopathic drugs the situation is very different. Their method of preparation is based essentially on two steps: sequential dilution with ‘succussion’ or ‘dynamisation’ (vigorous turbulent shaking). A molecular view of the matter and a trivial calculation demonstrates that, often it is extremely improbable that even one molecule of the compound present in the original solution persists in a vial of the final medicine. The role of succussion is not obvious, even less the diverse standards of methods of preparation.

Under the pressure of criticism, the natural evolution of researchers interested in finding acceptable scientific justifications of homeopathy has been to go from purely medical concepts of effective therapy to chemistry and finally to fundamental physics. Ultimately, schematically, the answer: if there is ‘only’ water in homeopathic medicines, then the explanation of the therapeutic action must be in pure water, itself!

This intellectual evolution is a paradox. While for many drugs, the action is known at a biological, sometimes at a chemical, but almost never at a physical level (that of the structure and energies defined with atomic resolution); for homeopathy, the discussion jumped directly into this microscopic sub-molecular physics world. The mixture of the precise methodology characterizing research in physics and procedures deriving from pharmacology in research in homeopathy is striking. For example, several measurements of physical properties of diluted solutions have been done double-blinded. An extreme and provocative hypothesis is that water can retain a ‘memory’ of substances previously dissolved in it.¹

A critical analysis of several publications shows that several issues remain open to question. Schematically, one can distinguish the following:

(1) How different from pure water are highly diluted solutions? In other words, is the simple calculation of the number of molecules of the ‘active principle’ per unit volume of the solution sufficient to account for the composition of homeopathic medicines?

(2) If succussion is an essential step in the preparation of homeopathic medicines, what is exactly its role? How does it influence the dilution procedure?

(3) What is the behaviour of complex molecules (eg biopolymers, organic compounds, surfactants, etc.) during the dilution process?

A clear answer to these (and perhaps other) questions is a necessary and essential precondition to any study of ‘pure’ water. Indeed, the conditions of preparation and conservation of homeopathic medicines are far from respecting the simplest procedures required in physical studies of pure water.

Some issues should be controlled more systematically:

(1) Pure water is a very powerful solvent of many substances. For example, it dissolves and forms specific bonds with silica. In contact with the surface of quartz, water forms stable silanol groups (Si–O–H). With time, silica molecules and silicon atoms are solubilised and hydrated. The number of these ‘impurities’ is huge as compared with the calculated amount of molecules of the starting substance in most homeopathic medicines.

It may be useful to recall that the interaction of water with solid surfaces is so strong that studies of nucleation must be done with minute amounts of water kept in levitation, without any contact with solid surfaces. The interaction with solid surfaces is so important that if a supercooled liquid freezes, it must be heated up to temperatures higher than the melting point in order to be supercooled again. Less important for water than for other liquids (eg gallium), this effect is due to more favourable nucleation of the solid form at the solid surface.

Another point deserving investigation is the storage of homeopathic solutions over long periods of time. This procedure is totally incompatible with a chemical purity of water, even at a modest level.

(2) The main consequence of succussion is the insertion of substantial amounts of air from the environment where the procedure takes place. In a laboratory that is not a cleanroom (such as those used for example in electronics), the procedure brings into the solution not only the gases present in the atmosphere (oxygen, nitrogen, argon,…) but also dust particles, micro-droplets of water, etc. Recent studies² show that the properties of solutions are drastically modified when succussion is done under different atmospheres or at different pressures, a fact which should encourage further studies in this direction.

(3) Many substances, which contain pharmacologically active principles, are not soluble in water. Some are previously diluted in alcohol suggesting the presence of surfactant molecules that go spontaneously to interfaces such as the free surface, the interface between the solution and micro-droplets of gases and the interface with the vial. Again, several very promising and striking studies performed by the analysis of the thermoluminescence of frozen solutions open new and exciting perspectives.³

To summarize, it is striking that in publications concerning highly diluted solutions, chemical ‘purity’ is assumed, solely on the basis of a calculation based on the dilution procedure itself. In fact most of the samples studied are far from being ‘pure water’. It would be interesting to perform to a real analysis of the composition of the solutions with physical methods such as mass spectroscopy.

Properties of liquid water

As stated above, many experiments with homeopathic medicines assume the purity of the highly diluted solutions and attribute its therapeutic action to modifications of the structure and dynamics of the pure liquid itself due to the past presence of a solute.¹ Such a strong hypothesis would imply not only general or random changes but also a large variety of changes, specific to each solute. The main purpose of this paper is to recall that this hypothesis is totally incompatible with our present knowledge of liquid water.

Water, in all its forms (crystal, liquid, gas and amorphous forms) is certainly the most studied of all substances. All its properties have been measured with extremely high accuracy in very different conditions, including metastable states and ‘extreme’ conditions. This is due to the central role of water in many scientific domains in physics, chemistry, geophysics and, of course, biophysics. Essentially all known experimental techniques and computer simulations have been used to precise details of the behaviour of water at scales extending from hydrodynamics to the nuclear and electronic levels. In other words, water is not an unknown substance!

However, do we know ‘everything’ about water? Certainly not: several puzzling questions are open to discussion. In brief, the main open question about pure water concerns the supercooled (metastable) state (ie liquid water at temperatures below its freezing point) and its relation with different amorphous (glassy) states. The structure of liquid water, at atmospheric pressure, is not known in a large temperature range extending from the vicinity of the temperature of homogeneous nucleation of ice (−42 °C) down to the temperature of the glass transition (−140 °C). This problem is the object of debate and speculation mostly based in extrapolations of simulations of molecular dynamics performed by computer.^{[4] and [5]}

Another important domain of research is ‘confined water’, ie water occupying extremely small volumes, for example, in porous materials, in thin layers or in small pools formed at hydrophobic sites of bio-molecules. In this case, there is a large variety of situations that depend essentially on the nature of the substrate and on the relative importance of the number of molecules at the surface and in the bulk of the small volume. However, pure water at ambient conditions is well understood. Let us review some of its main properties that may be related to the subject of this paper.

Water is a simple molecule containing three atoms: one of oxygen and two of hydrogen strongly bound by covalent bonds. Because of the hybridisation of the molecular orbitals, the shape of the molecule is a V with the oxygen occupying the vertex of an angle of 104°; the O–H distance is almost exactly 0.1 nm. When two water molecules are sufficiently close, they orient one against the other to establish a chemical bond, called hydrogen bond. In this bond, one hydrogen atom is shared by two neighbouring molecules (Figure 1). The bonding energy is about 10 times larger than the kinetic energy but the bond is ‘fragile’ due the vibratory motions of the hydrogen atom particularly in the direction perpendicular to the line O–HO. It is possible to measure accurately the typical time for which the three atoms are aligned (the lifetime of hydrogen bonds): it is of the order of 0.9 ps (9×10⁻¹³ s) at room temperature.

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Figure 1. Schematic representation of a hydrogen bond in water. The large circles represent two oxygen atoms of neighbouring molecules; the small circle is the hydrogen atom attached to the oxygen on the left hand side by a covalent bond. The length of the hydrogen bond is 0.18 nm.The hydrogen atom vibrates in all directions. Vibrations perpendicular to the bond are most likely to break the bond.

Because of its geometry, a water molecule can easily form four hydrogen bonds with four neighbouring molecules. This corresponds to the structural arrangement in common ice (I_h or hexagonal form). The angle of 104° is sufficiently close to the tetrahedral angle (109°) to impose this very open structure where each molecule is surrounded by four others at the apex of a tetrahedron (Figure 2). In liquid water this local geometry exists partly: on average a water molecule has 4.5 neighbours but this number decreases with decreasing temperature because the average number of ‘intact’ bonds increases. Incidentally, it is this decrease of the number of first neighbours that explains why the density of water decreases at low temperatures. At 4 °C, which is the temperature of maximum density, this effect compensates that of thermal expansion.

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Figure 2. Tetrahedral arrangement of five water molecules. The vibrational motion of a hydrogen atom is represented by an arc on the right-hand side of the figure (adapted from G Walrafen).

The average number of ‘intact’ bonds at a given moment is relatively high, although lower than in alcohols, for example. It is of the order of 60% which justifies seeing liquid water as a 3-dimensional network of hydrogen bonds, like a gel. But a gel with a life time of 1 picosecond (ps)! This means that in an ‘instantaneous picture’ of water structure (possible to obtain by computer simulations) one can identify local structures such as rings of 5, 6 or 7 molecules, regions with higher density of bonds than others, etc. All these structural properties can be identified by several techniques and correspond to thermodynamic properties. For example, the increase of isothermal compressibility observed at low temperatures is due to the enhancement of density fluctuations. It is very important to note that such fluctuations are not due to aggregation or formation of clusters. Hydrogen bonds form and break very rapidly generating short lived fluctuations of local density. In other words, even if at a given moment one can identify a region of higher density than the average, it will disappear after a very short time and will appear statistically in another place without any form of coherent motion such as would exist if a cluster was diffusing inside the liquid.

Historically, the first models of liquid water (due to WC Roentgen) represented liquid water as a mixture of an ideal liquid and small ice-like clusters. This model has been ruled out by many experiments. Among them, small angle X-ray scattering eliminates unambiguously any possibility of existence of clusters or aggregates in liquid water, even at very low temperatures.^{[6] and [7]}

Isolated or confined water molecules can have their mobility totally restricted. In such cases, the lifetime of a hydrogen bond can be infinite. This situation is frequent in proteins where hydrogen bonds with water can play a central role in protein structure. But, in these situations, water molecules don’t constitute a liquid. Consequently, it is worth emphasizing that to postulate the existence of stable structures in pure water is totally wrong. This is one of the limits imposed by the knowledge of the structure of water.

Aqueous solutions

In aqueous solutions, the situation is more diverse. Water can dissolve and mix with many substances in different proportions (salts, acids, various alcohols, sugars, and gases, etc). Both local structure and dynamic properties may be drastically modified. Two well known examples give an idea of the diversity of situations. Trehalose is a sugar that promotes the formation of glassy water even when extremely dilute. It is present in animals and plants which, because of this property, can survive very low temperatures. Other examples are aerogels of silica with a huge content of water, which can contain more than 95% water while remaining macroscopically solid.

Generally speaking, the inclusion of molecules or ions destroys local tetrahedral geometry. Depending on the nature of the compound, the molecules of water arrange in a large variety of local structures. For example, when a salt is dissolved in water, it is dissociated into two ions each of which is surrounded by a layer of hydration where the strong electrostatic interactions between the charge of the ion and the dipoles of water generate a mini-cluster (Figure 3). The life time of this cluster is 10 to 100 times longer than the lifetime of hydrogen bonds but is not infinite, because of the exchange between molecules of water in the hydration shell and those of the bulk.

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Figure 3. Schematic representation of the arrangement around an anion (left) and a cation (right). In the first case the dipolar moment of the water molecules is directed towards the ion; in the opposite direction in the case of the anion. The screening of the electrical field of the ions is very efficient and the structure of water beyond the first hydration layer is almost not modified.

However, it is erroneous to believe that the electrical field generated by the ions extends over large distances. Actually, it is screened by the hydration layer. There is a large literature about the structure in hydration shells. The number of water molecules, distances and angles are known with great accuracy from neutron scattering experiments based on isotopic substitution.⁸

A very different situation concerns the solubility of hydrophobic atoms and molecules, such as methane or noble gases. In this case, water has tendency to form clathrate-like structures around the solute. A clathrate is a polyhedral structure; frequently a dodecahedron with pentagonal faces. This is a very stable structure, because the internal angle of the pentagon (104°) is equal to the internal angle, HOH, of the molecule. It forms a cage and the prisoner is the hydrophobic solute. The short lifetime of hydrogen bonds does not allow the formation of stable or long-lived clusters. Experiments simply detect, at best, a tendency to the formation of short lived planar pentagons.

Finally, it is interesting to consider situations in which stable aggregates are formed. The most interesting, including many industrial applications, are surfactants, which are molecules with a hydrophilic head (sometimes polar) and one or two hydrophobic tails. When dissolved in water in sufficiently large amount (above a critical micellar concentration, c.m.c.) they form structured clusters called micelles (Figure 4). The heads are at the external surface and the hydrophobic tails minimise the interaction energy with water inside the sphere. These structures are very stable. They persist essentially for ever, even if there are many exchanges of surfactant molecules between micelles, either by diffusion or as a result of collisions. Many structures of this type are known, of different sizes and shapes. Some are very important in biology or in pharmacy. For example, bi-layers of phospholipids mimic quite well some physical properties of biologic membranes, and vesicles are sometimes used as vectors or carriers of drugs.

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Figure 4. Spherical micellar aggregate showing the hydrophilic heads in contacts with the surrounding water or aqueous solvent. The hydrophobic tails fill the internal part of the spherical droplet.

In small quantities, surfactant molecules migrate to interfaces in geometries that minimise the interaction between the tails and water. Even at very low concentration they can modify substantially the surface tension of water. Whenever surfactant molecules are present in a substance, one must take into account their specific interactions with water.

Conclusion

To summarize this short overview, one can say that water is a ‘complex’ liquid with many fascinating, sometimes unique aspects. Except for some academic aspects concerning supercooled water, the structure of the liquid is well known. In particular, it is certain that:

(a) There are no water clusters in pure liquid water, but only density fluctuations.

(b) The longest life of any structure observed in liquid water is of the order of 1 ps (10⁻¹² s).

This is why any interpretation calling for ‘memory’ effects in pure water must be totally excluded.

In contrast, there is great variety of behaviour of solutes depending on many parameters. Even in small quantities, some solutes can modify substantially some properties of pure water. Special attention should be given to surfactants, sugars and polymeric substances. Since homeopathic medicines are prepared in ‘extremely high dilutions’ but following a procedure that does not produce necessarily extremely pure water, experiments should address the problem of the presence of minute amounts of solutes as has recently been done recently, with striking results.²

Otherwise, as stressed at the beginning, the advantages of homeopathic treatments should be taken at a medical level, which, after all, is the case for other drugs recognized for their remarkable although not yet explained effectiveness.

References

1 E. Davenas, F. Beauvais and J. Amara et al., Human basophil degranulation triggered by very dilute antiserum against IgE, Nature 333 (1988), pp. 816–818. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

[2] L. Rey, Can low temperature Thermoluminescence cast light on the nature of ultra-high dilutions?, Homp 96 (2007), pp. 170–174. SummaryPlus | Full Text + Links | PDF (267 K)

3 L. Rey, Thermoluminescence of ultra-high dilutions of lithium chloride and sodium chloride, Physica A 323 (2003), pp. 67–74. SummaryPlus | Full Text + Links | PDF (306 K) | View Record in Scopus | Cited By in Scopus

4 O. Mishima and H.E. Stanley, The relationship between liquid, supercooled and glassy water, Nature 396 (1998), pp. 329–335. View Record in Scopus | Cited By in Scopus

5 J. Teixeira, A. Luzar and S. Longeville, Dynamics of hydrogen bonds: how to probe their role in the unusual properties of liquid water, J. Phys.: Cond. Matter 18 (2006), pp. S2353–S2362. Full Text via CrossRef

6 R.W. Hendricks, P.G. Mardon and L.B. Schaffer, X-ray zero-angle scattering cross section of water, J. Chem. Phys. 61 (1974), pp. 319–322. Full Text via CrossRef

7 L. Bosio, J. Teixeira and H.E. Stanley, Enhanced density fluctuations in supercooled H₂O, D₂O and ethanol–water solutions: evidence from small-angle X-ray scattering, Phys Rev Lett 46 (1981), pp. 597–600. Full Text via CrossRef

8 L. Friedman H, A Course in Statistical Mechanics, Prentice Hall College Div. (1985).

Correspondence: Laboratoire Léon Brillouin (CEA/CNRS), CEA Saclay, 91191 Gif-sur-Yvette Cedex, France.

Homeopathy
Volume 96, Issue 3, July 2007, Pages 158-162
The Memory of Water

This is part of the Homeopathy journal club described here:

www.badscience.net/?p=490

doi:10.1016/j.homp.2007.05.007    
Copyright © 2007 Elsevier Ltd All rights reserved. The ‘Memory of Water’: an almost deciphered enigma. Dissipative structures in extremely dilute aqueous solutions

V. Elia^{1, ,} , E. Napoli¹ and R. Germano^2
1Dipto. di Chimica, Università ‘Federico II’ di Napoli, Complesso Universitario di Monte S.Angelo, via Cintia, 80126 Napoli, Italy;
²PROMETE Srl – INFM Spin off Company, Via Buongiovanni 49, San Giorgio a Cremano, 80046 Napoli, Italy
Received 2 April 2007;  revised 22 May 2007;  accepted 29 May 2007.  Available online 31 July 2007.

In the last decade, we have investigated from the physicochemical point of view, whether water prepared by the procedures of homeopathic medicine (leading inexorably to systems without any molecule different from the solvent) results in water different from the initial water?

The answer, unexpectedly, but strongly supported by many experimental results is positive. We used well-established physicochemical techniques: flux calorimetry, conductometry, pHmetry and galvanic cell electrodes potential. Unexpectedly the physicochemical parameters evolve in time.

The water solvent exhibits large changes in measurable physicochemical properties as a function of its history, the solute previously dissolved, and time. In particular we found evidence of two new phenomena, both totally unpredicted, in homeopathic dilutions: the presence of a maximum in the measured physicochemical parameters vs sample age, and their dependence on the volume in which the dilution is stored. These new experimental results strongly suggest the presence of an extended and ‘ordered’ dynamics involving liquid water molecules.

Keywords: homeopathy; calorimetry; conductivity; pH; dissipative structures

Article Outline

Introduction

Methods

Ageing effects

Conclusion

References

Introduction

The ‘Memory of Water’ is a journalistic expression, first used in the French newspaper Le Monde, after the publication in 1988 of Jacques Benveniste’s famous paper in the international scientific journal Nature.¹ In this paper he claimed, with biological experimental data, that ‘homeopathic dilutions’ of substances (ie so much diluted as to not contain any molecules of the substance initially diluted in it) are able to induce biological effects typical of the substance initially dissolved in it. The ‘memory of water’ is a synthesis of a still unexplained phenomenon. Recent scientific publications suggest some possible ways to experimentally validate the reality of a whole new class of physicochemical new phenomena concerning liquid water.²

It seems that it really is possible to obtain physicochemical information depending on the recent or remote ‘history’ of a water sample (in Prigogine’s terminology: breaking of the temporal symmetry), almost as in the better known case of magnetic materials (Prigogine would say: breaking of the spatial symmetry).³ The so-called memory of water, is connected to the capacity of this kind of solvent, a multi-variable complex system, to be influenced by very tiny perturbations, such as mechanical or electromagnetic actions, in such a way to move away from the initial equilibrium conditions, and this is increasingly established. The ‘memory of water’, in this sense, is comprehensible in the framework of the theory of Irreversible Processes Thermodynamics due to the Nobel Laureate for Chemistry (1977), Ilya Prigogine.³

In the last 10 years,^{[4], [5], [6], [7], [8], [9], [10], [11], [12] and [13]} our research group has investigated this problem from the point of view of the physicochemical properties of water when prepared following the procedures of homeopathic medicine preparation: iterative dilutions (of specific solutes of medical interest) followed by agitation (succussion). This method leads inexorably to systems without any molecule different from the solvent, in our case pure water.

Can the ‘new water’ thus obtained really be ‘different’ from the initial one? Answering this question was our challenge. The answer, unexpected but strongly supported by the experimental results, is affirmative. In the meantime, other research groups came to similar conclusions using different experimental models and other methodologies.^{[14], [15], [16], [17], [18], [19] and [20]} We also want to note here Giorgio Piccardi, the founder of the Italian physical-chemistry, and his pioneering work concerning fluctuating chemical reactions.^{[21], [22], [23] and [24]} A critical mass of experimental data² necessary to evidence a new class of physicochemical phenomena of the water has now been reached.

Methods

The experimental methodologies used for our investigations were chosen as the most efficient among the many tested. We list them, without entering into the technical details, but emphasising that they are well-established physicochemical methodologies: flux calorimetry, conductometry, pHmetry and galvanic cell electrode potential. The greatest difficulty of the preliminary work, which lasted for many years, was the selection of the most enlightening experimental methodologies and the establishment of optimal experimental conditions. It was also difficult to evaluate the contribution of the impurities released by the glassware, to the measured experimental values. In fact, the problem of impurities has been the principal objection probably due to the strong prejudice against the possibility that the procedures followed might really change the physicochemical nature of water.

Figure 1, Figure 2 and Figure 3 show that the presence of impurities released by the glassware makes a significant contribution to the physicochemical state of the dilutions, but it is not relevant in comparison with the unexpected contribution (much higher than the range of the experimental errors) of the auto-organisation process of the water molecules-the water is far from the thermodynamic equilibrium (see below) –and this auto-organization is triggered by external perturbations (such as iterative dilution and succussion).

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Figure 1. Heat of mixing, Q_mix, vs concentration (determined by analytic methods) of impurities, M_tot, released by the glass vessels. Black symbols: heat of mixing of homeopathic solutions with sodium hydroxide, NaOH, 0.01 M (mol kg⁻¹); red line: heat of mixing of aqueous solutions containing the same amount of impurities determined in the homeopathic solutions. The absolute values of the heat of mixing with sodium hydroxide using homeopathic solutions are always higher than the corresponding heat of mixing determined only by the ‘chemical’ contribution originating from the glassware.

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Figure 2. Specific conductivity, χ, vs concentration (determined by analytic methods) of impurities, M_Na+, released by the glass vessels. Black symbols: specific conductivity of the homeopathic dilutions; red line: specific conductivity of aqueous solution containing only the same amount of impurities determined in the homeopathic solutions.

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Figure 3. pH values vs concentration of impurities M_T (determined by analytic methods), released by the glassware. Circle symbols: pH of homeopathic dilutions; triangle symbols: pH of aqueous solution containing the same amount of impurities determined by analytic methods in the homeopathic solutions.

Figure 1, Figure 2 and Figure 3 show: (i) how the contributions of impurity were taken into account; (ii) the major contribution of ‘something’ different from any possible substance of chemical origin. This is a preliminary result but it cleared misunderstandings from the experimental methodologies and allowed us to proceed to collect further information and insights on the nature of the ‘homeopathic dilutions’.^{[4], [5], [6], [7], [8], [9], [10], [11], [12] and [13]}

It is important to emphasise that, from the studies so far conducted, we cannot derive reproducible information concerning the influence of the different degrees of homeopathic dilution or the nature of the active principle (solute) on the measured physicochemical parameters. For this reason the experimental data reported in the figures are not given in terms of homeopathic dilutions or name of homeopathic medicine.

Ageing effects

A stimulating, and somewhat serendipitous, result very important in understanding the complex system under study, was that the physicochemical properties of the homeopathic solutions depend on time. The fact that the numerous experiences were performed over many years, naturally introduced the time parameter. The analysis of the experimental results vs the ‘arrow of time’ was of unexpected relevance,^{[7], [8], [9] and [10]} and led to the idea that the system under observation (homeopathic solution) is a closed system (able to exchange only energy with the external environment), far from thermodynamic equilibrium, which allows structures with a local order higher than the water around them to emerge from chaos (‘dissipative structures’).

Figure 4 and Figure 5 show that, unexpectedly, the investigated physicochemical parameter increases with time. In other words, ageing modifies the physicochemical nature of homeopathic solutions.

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Figure 4. Heat of mixing, Q_mix, vs the samples age, t, for six homeopathic solutions.

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Figure 5. Excess specific conductivity, χ^E (μS cm⁻¹) (defined as the experimental difference between the experimental χ value and the contribution to this parameter by the presence of impurities (χ^chem) vs the samples age, t, for six homeopathic solutions. Each studied sample has its own peculiar χ^E vs time evolution but with overall similar behaviour: an increment of χ^E in time.

What is the interpretation of this newly-observed characteristic of the homeopathic solutions? Are we simply observing a system seeking an energetic minimum and a new equilibrium in a slow kinetic process or this is something totally different? From the data in Figure 4 and Figure 5 we deduce that the temporal variations of the reported parameters are very slow, because it takes many months to evidence them unambiguously. However, this temporal behaviour does not match the idea of a simple slow kinetic. In fact, following the reductio ad absurdum, if there exists an energetic minimum towards which the system could move, it would be impossible that in the time that water has existed, it has not reached this hypothetical minimum. Moreover for both parameters (specific conductivity and heat of mixing with alkaline solutions) an increase with time is observed. The correlation between the specific conductivity and the heat of mixing with alkaline solutions, shown in Figure 6 is linear; in other words, these two parameters have the same underlying cause. This result provoked us to investigate the nature of the mechanism able to increase simultaneously the electrical conductivity and the heat of mixing with alkaline solutions, after repeated dilutions and succussions. The descriptive model proposed below, although simple, is in agreement with the experimentally observed ageing effect.

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Figure 6. Specific conductivity, χ, vs excess heat of mixing, Q_mix^E (the difference between the experimental Q_mix value and the contribution to this parameter due to the presence of impurities), for a given volume of homeopathic solution.

An explanation of the electrical conductivity increasing after the preparation procedure and ageing may be based on the so-called ‘hopping mechanism’, proposed by C.J.T. Grotthuss (1806)²⁵ to explain the much higher mobility (about 5 times) of H⁺ and OH⁻ ions (always present in liquid water) in comparison with other ions of comparable ionic radius. If H₂O molecular clusters are present in the solution, bonded by hydrogen bonds, the hydrogen ions H⁺ colliding them experience the ‘hopping’ phenomenon (Figure 7): the water molecules catch an H⁺ ion at one end of the cluster (for the sake of simplicity considered linear) and release instantaneously another H⁺ ion at the other end of the cluster. The drift velocity under an electrical potential gradient (a measure of the conductivity) is much increased in comparison with that of ions which do not encounter H₂O molecular clusters. The greater the number of the clusters and/or their length, the higher the conductivity value. The correlation between the electrical conductivity and the heat of mixing with alkaline solutions is a consequence of H₂O clusters breaking, due to the pH variation (see Figure 8).

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Figure 7. Schematic representation of the Grotthuss hypothesis of the proton (H⁺) hopping mechanism to explain the much higher mobility (defined as the ionic drift velocity under a unitary gradient of electrical potential V cm⁻¹) of H⁺ and OH⁻ ions in water (H₂O=H⁺+OH⁻) in comparison with other ions of comparable ionic radius.

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Figure 8. Schematic representation of the phenomenon of molecular clusters breaking, due to pH variation during the experimental procedure of determining the heat of mixing with hydroxide solutions (NaOH) 0.01 M (mol kg⁻¹ ) in a calorimetric cell. The experimental procedure consists of mixing a homeopathic dilution (that we suppose richer in H₂O molecular clusters than the ‘standard’ water solvent) with an alkaline solution. The pH variation seems to reflect breaking of hydrogen-bonded H₂O clusters, determining a transition order→disorder. This is experimentally evidenced by the increased heat of mixing compared to ‘normal’ water containing few molecular clusters.

The greater the number of the clusters and the larger their dimensions, the more is the measured thermal effect (Figure 8). These two experimental phenomena witness the same thing, both are sensitive to the number and/or dimensions of the clusters.

Let us return to the question: Are we measuring the presence of stable clusters seeking an energetic minimum? Or of unstable clusters consisting of dissipative auto-organised structures that are far from equilibrium and which remain or move away from equilibrium as a function of their ability to exchange energy with the external environment? We have already emphasised that the hypothesis of systems slowly evolving towards new equilibrium states is not compatible with our experimental findings. In particular, the hypothesis of systems evolving towards a minimum, even very slowly, contrasts with two new and very unexpected experimental phenomena characterising homeopathic dilutions:

(a) the presence of a maximum in the physicochemical parameters with sample age (Figure 9);

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Figure 9. Specific excess conductivity, χ^E, vs the samples ageing, t. Each curve describes the temporal evolution of Arnica Montana (AM) samples in homeopathic dilutions prepared from the same mother tincture. There is no specific correlation between the χ^E behaviour and the degree of dilution (CH) of the samples.

(b) the dependence of the physicochemical parameters (apart from age) also on the volume in which the homeopathic dilution is stored (Figure 10).

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Figure 10. Excess specific conductivity, χ^E, vs ageing volume, V. Each point represents the value of χ^E for each single dilution, experimentally determined at the same age. There is a very strong variation of the parameter, about one order of magnitude, for the systems aged in very small volumes. This volume dependence cannot be explained in the frame of the classical physico-chemistry.

Phenomenon (b) is absolutely anomalous and inexplicable in the current paradigm,¹³ it appears to be in sharp contradiction with the classical concept that an intensive physical quantity cannot depend on the volume.

The temporal evolution of the excess specific conductivity of four sample systems is shown in Figure 11. The samples were obtained as follows: a highly diluted aqueous system was divided into three smaller volumes at a certain ‘age’. As the figure shows, the excess specific conductivity (χ^E) behaviour across time of small volume samples is very different from that of larger volume samples. The larger volume sample does not display relevant modifications across time, while each new system of smaller volume evolves in a different way, with an overall common behaviour characterised by the presence of a maximum. This means that the evolution over time depends on the initial state (in this case: large or small volume), in a sense the systems have a ‘memory’ of the initial conditions.

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Figure 11. Excess specific conductivity, χ^E, vs sample age, t. In this experiment, a homeopathic dilution of Arnica Montana was left to age for about 250 days in a volume of about 200 ml. At this time point 18 ml were removed and divided into three different vessels of equal shape, containing 10, 5 and 3 ml. The four obtained samples, 182, 10, 5 and 3 ml, were studied vs time. Their temporal evolution was dramatically influenced by the perturbation induced by the repartition into smaller volumes. In particular the higher volume of 182 ml did not experience particular temporal variations, while in the case of the smaller volumes, a large temporal evolution was observed, depending strongly on the starting point.

Another example of such ‘memory’ of the system is apparent in the experimental data displayed in Figure 12, which shows the temporal evolution of the excess specific conductivity for samples made from the same mother tincture diluted in double distilled water without succussion, in different dilution ratios.¹² Again, the system’s evolution in time is strongly conditioned by the initial conditions, with temporal variations characterised by very different maximum and slope values: past history influences the evolution of the ‘pure water’ system.

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Figure 12. Excess specific conductivity, χ^E, vs age, t, of samples obtained by a simple dilution of the ‘mother tincture’ with double distilled water without succussion, in different dilution ratios (r). The volumes of the studied solutions were the same. The temporal evolution of the various systems, perturbed only by the simple dilution without succussion, is strongly dependent by the new starting state. In particular, the system with dilution 1:1, r=0.5 (final volume is twice the initial one) exhibits an initial χ^E value markedly lower than the solution from which it was obtained, then, in about 45 days, exhibiting χ^E values much higher with respect to the ‘mother tincture’, reaching a sharp maximum. In this case, the applied perturbation, determines a strongly different starting point, as well as different temporal evolution.

The apparent contradiction between the concept of intensive quantity, such as specific conductivity and heat of mixing, and the experimental evidence of dependence on volume may be solved by considering that, within the solutions there are molecular clusters consisting of water molecules connected by hydrogen-bonds, in far from equilibrium conditions. They can remain in, or move away, from their unstable equilibrium state, dissipating energy derived from the external environment: they are ‘dissipative structures’ as described by Prigogine.³

The spontaneous formation of molecular clusters in water is foreseen by the Coherent Quantum Electrodynamics (Coherent QED) without introducing the existence of hydrogen-bonds. This theoretical formulation, due to G. Preparata, E. Del Giudice, et al predicts the physicochemical properties of the water,^{[2], [26], [27], [28] and [29]} much better than other theories. The introduction of the ‘arrow of time’ into this theoretical framework should yield very interesting results.

Conclusion

We propose a simplified empirical model that in principle seems able to explain the unexpected dependence of the physicochemical parameters on the volumes used.

A first hypothesis to explain the experimental results is to suppose that the solutions, after strong agitation (succussion), enter a far from equilibrium state, remaining there or getting even farther by dissipating energy in the form and amount necessary to stay in the far from equilibrium state. Then, assuming that radiant energy is exchanged, we can further suppose that, for a given flux of dissipated energy (W cm⁻²), the same number of dissipative structures would be formed, even if contained in different volumes. In this frame, on average, at any given age, small volumes of water will contain a higher ‘concentration’ of dissipative structures in comparison with larger volumes (Figure 13). The physicochemical parameters electrical conductivity and heat of mixing are in fact functions of the number, size and shape of the dissipative structures.

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Figure 13. Schematic representation of a possible temporal evolution of a homeopathic solution, showing a variation of intensive quantities such as χ^E (μS cm⁻¹) and Q^E_mix (J kg⁻¹) vs the storage volume. At the time zero, the two vessels, of equal shape and volume, contain two identical homeopathic dilutions (same age, same active principle, same CH dilution) with no experimentally measurable effects determined by dissipative structures, because of their small number (Figure 13a). Assuming the same formation efficiency (and/or increase in size) for the dissipative structures in the two systems (small volume and large volume), with the same conditions of energetic flux, the number and/or size of the dissipative structures is almost the same in the two containers, at any given time (Figure 13b and c). So, when dissipative structures are numerically increasing, their concentration is much higher in the small volume than in the large one. Consequently, intensive quantities such as those measured, χ^E (μS cm⁻¹) and Q^E_mix (J kg⁻¹), sensitive to the structure concentration, will show a temporal behaviour dependent on the volume.

We conclude the following:

• the parameters whose values results ‘in excess’ (in general: variable with the history of the solvent in time) are correlated with the dynamics of supermolecular (mesoscopic) structures in the water solvent;

• the temporal evolution of the parameters is not connected to the tendency to seek an energetic minimum;

• an empirical interpretation, consistent with all current experimental data, is based on the presence of dissipative structures.

Succussion may be the trigger for the spontaneous formation of dissipative structures, that is the emergence of new dynamics. The temporal evolution may be connected to the variation of the number, dimension or the shape of the dissipative structures. It is well known, in Thermodynamics of Irreversible Processes, that the temporal evolution of the systems depends on the initial conditions and on the way the systems evolve.Much new experimental data converge towards the validation of the statement that water, at least in the context of the procedure of the homeopathic medicine production, really has a ‘memory’. That is to say: the water solvent shows experimentally measurable physicochemical properties that vary as a function of the ‘lived path’, of the solute previously dissolved, and of elapsed time.

Without doubt liquid water has an extended and ‘ordered’ dynamics involving the whole body of the liquid. It is much more complex than the normal idea of a banal and chaotic cluster of ‘molecular balls’.

References

1 E. Davenas, F. Beauvais and J. Amara et al., Human basophil degranulation triggered by very dilute antiserum against IgE, Nature 333 (1988), pp. 816–818. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

2 R. Germano, AQUA. L’acqua elettromagnetica e le sue mirabolanti avventure, Bibliopolis, Napoli (2007).

3 I. Prigogine, From Being to Becoming. Time and Complexity in the Physical Sciences, Freeman, San Francisco (1980).

4 V. Elia and M. Niccoli, Thermodynamics of extremely diluted aqueous solutions, Ann NY Acad Sci 879 (1999), p. 241. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

5 V. Elia and M. Niccoli, New physico-chemical properties of water induced by mechanical treatments. A Calorimetric study at 25 °C, J Therm Anal Calorimetry 61 (2000), pp. 527–537. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

6 V. Elia and M. Niccoli, New Physico-chemical properties of extremely diluted aqueous solutions, J Therm Anal Calorimetry 75 (2004), pp. 815–836. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

7 V. Elia, S. Baiano and I. Duro et al., New and permanent physico-chemical properties of the extremely diluted aqueous solutions of the homeopathic medicine. A conductivity measurements study at 25 °C in function of the age of the potencies, Homeopathy 93 (2004), pp. 144–150. SummaryPlus | Full Text + Links | PDF (154 K) | View Record in Scopus | Cited By in Scopus

8 V. Elia, E. Napoli and M. Niccoli et al., New physico-chemical properties of extremely diluted aqueous solutions. A calorimetric and conductivity study at 25 °C, J Therm Anal Calorimetry 78 (2004), pp. 331–342. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

9 V. Elia, M. Marchese and M. Montanino et al., Hydrohysteretic phenomena of ‘extremely diluted solutions’ induced by mechanical treatments. A calorimetric and conductometric study at 25 °C, J Solution Chem 34 (8) (2005), pp. 947–960. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

10 V. Elia, L. Elia and P. Cacace et al., Extremely diluted solutions as multi-variable systems. A study of calorimetric and conductometric behaviour as function of the parameter time, J Therm Anal Calorimetry 84 (2) (2006), pp. 317–323. View Record in Scopus | Cited By in Scopus

11 V. Elia, L. Elia and M. Marchese et al., Interaction of ‘extremely diluted solutions’ with aqueous solutions of hydrochloric acid and sodium hydroxide. A calorimetric study at 298 K, J Mol Liq 130 (2007), pp. 15–20. SummaryPlus | Full Text + Links | PDF (189 K) | View Record in Scopus | Cited By in Scopus

12 V. Elia, L. Elia and M. Montanino et al., Conductometric studies of the serially diluted and agitated solutions. On an anomalous effect that depends on the dilution process, J Mol Liq 135 (2007), pp. 158–165. SummaryPlus | Full Text + Links | PDF (235 K)

13 V. Elia, L. Elia and E. Napoli et al., Conductometric and calorimetric studies of serially diluted and agitated solutions: the dependence of intensive parameters on volume, Int J Ecodyn 1 (4) (2006), pp. 1–12.

14 P. Belon, J. Cumps and P.F. Mannaioni et al., Inhibition of human basophil degranulation by successive histamine dilutions: results of a European multi-centre trial, Inflammation Research 48 (Suppl 1) (1999), pp. S17–S18. View Record in Scopus | Cited By in Scopus

15 L. Rey, Thermoluminescence of ultra-high dilutions of lithium chloride and sodium chloride, Physica A 323 (2003), pp. 67–74. SummaryPlus | Full Text + Links | PDF (306 K) | View Record in Scopus | Cited By in Scopus

16 L. Betti, M. Brizzi and D. Nani et al., A pilot statistical study with homoeopathic potencies of arsenicum album in wheat germination as a simple model, Br Hom J 83 (1994), pp. 195–201. Abstract | PDF (432 K)

17 L. Betti, M. Brizzi and D. Nani et al., Effect of high dilutions of arsenicum album on wheat seedlings from seed poisoned with the same substance, Br Hom J 86 (1997), pp. 86–89. Abstract | PDF (276 K)

18 M. Brizzi, D. Nani and M. Peruzzi et al., The problem of homoeopathy effectiveness: a comparative analysis of different statistical interpretations of a large data collection from a simple wheat germination model, Br Hom J 89 (2000), pp. 1–5.

19 P. Torrigiani, A.L. Rabiti and C. Bortolotti et al., Polyamine synthesis and accumulation in the hypersensitive response to TMV in Nicotiana tabacum, New Phytol 135 (1997), pp. 467–473. View Record in Scopus | Cited By in Scopus

20 A.L. Rabiti, L. Betti and C. Bortolotti et al., Short term polyamine response in TMV-inoculated hypersensitive and susceptible tobacco plants, New Phytol 139 (1998), pp. 549–553. View Record in Scopus | Cited By in Scopus

21 G. Piccardi and R. Cini, Polymerization and the low-frequency electromagnetic field, J Polym Sci 48 (1960), p. 393. Full Text via CrossRef

22 G. Piccardi, Chemical test made in Antarctic, Geofis Meteorol XII (1963), p. 55.

23 G. Piccardi, 22 year solar cycle and chemical test, Geofis Meteorol XX (1961), p. 104.

24 F. De Meyer and C. Capel-Boute, Statistical analysis of Piccardi chemical tests, Int J Biometeorol 31 (1987), pp. 301–322.

25 C.J.T. Grotthuss, Sur la décomposition de l’eau et des corps qu’elle tient en dissolution à l’aide de l’électricité galvanique, Ann Chim 58 (1806), pp. 54–73.

26 E. Del Giudice, R. Mele and G. Preparata, Dicke Hamiltonian and superradiant phase transitions, Mod Phys Lett B 7 (28) (1993), pp. 1851–1855.

27 G. Preparata, QED Coherence in Matter, World Scientific, Singapore (1995).

28 R. Arani, I. Bono and E. Del Giudice et al., QED coherence and the thermodynamics of water, Int J Mod Phys B 9 (1995), p. 1813. Full Text via CrossRef

29 E. Del Giudice and G. Preparata, A new QED picture of water: understanding a few fascinating phenomena. In: E. Sassaroli et al., Editors, Macroscopic Quantum Coherence, World Scientific, Singapore (1998), pp. 49–64.

Correspondence: Vittorio Elia, Dipto. di Chimica, Università ‘Federico II’ di Napoli, Complesso Universitario di Monte S.Angelo, via Cintia, 80126 Napoli, Italy.

Homeopathy
Volume 96, Issue 3, July 2007, Pages 163-169
The Memory of Water

This is part of the Homeopathy journal club described here:

www.badscience.net/?p=490

doi:10.1016/j.homp.2007.05.004
Copyright © 2007 Elsevier Ltd All rights reserved. Can low-temperature thermoluminescence cast light on the nature of ultra-high dilutions?

Louis Rey^{, a,
a}Chemin de Verdonnet 2, CH-1010 Lausanne, Switzerland
Received 2 May 2007; revised 8 May 2007; accepted 16 May 2007. Available online 31 July 2007.

Abstract

Low-temperature thermoluminescence has been used in attempt to understand the particular structure of ultra high dilutions. Samples are activated by irradiation after freezing at the temperature of liquid nitrogen (77°K). Experimental results show that, in the course of rewarming, the thermoluminescent glow is susbtantially different between dilutions of different substances. It is suggested that the dispersed gas phase might play a role in this process.

Keywords: irradiation; frozen dilutions; nanobubbles; low-temperature glow

Article Outline

Introduction

Research objective

Method

Results

New prospects

Acknowledgements

References

Introduction

No chemical is more common on earth than water: it covers 75% of the earth’s surface with a total mass of 1.4 billion megatons. A very simple molecule, with one central, negatively charged, oxygen atom and two positively charged hydrogen atoms 0.1 nm apart at an angle of 104°¹ water is, nevertheless, a most atypical compound. In the liquid state, it is an abnormal fluid which should be a gas by comparison with other similar chemicals. Among other unusual properties, it increases in volume when crystallizing into solid ice at 0°C and boils at 100°C: both these temperatures are abnormally high for a substance which is neither a metal nor an ionic compound. Its dielectric constant as well as its increasing fluidity with rising pressure is equally odd.

In fact, liquid water is not a simple association of independent molecules; the molecules are actively interconnected by hydrogen bonds^{[2] and [3]}. Liquid water is, indeed, a structured fluid which behaves as a polymer. In an ever-moving universe, individual water molecules link to each other, most often in tetrahedral geometry, building evanescent clusters which are continuously formed and dissociated again at random in a pico-second timeframe. When an ionic compound is dissolved in H₂O, each ion is immediately surrounded by a spherical shell of water molecules so intensely that, should the concentration of the solute be high enough (over about 10%) all the shells come into contact and there is no more truly liquid water.

It can, thus, be understood that, in the preparation of an homeopathic medicine, any compound dispersed in water gives rise, from the outset, to a specific structure. When successive dilutions are made the violent turbulence created in the liquid by each succussion, helps to both maintain and possibly spread the original structure despite, progressively, the solute content of the dilution dropping by a factor of 100 with each centesimal step. However, Brownian motion is still very active and these ‘remnant structures’ fade away and reconstitute continuously. In other terms, we could say that homeopathic dilutions are ‘statistically structured’ and could remain so beyond the Avogadro number. Succussion appears to be an essential part of the overall process.

Research objective

It is easy to understand why, based upon this succession of dilutions–succussions, many scientists believe that eventually—and definitely beyond the Avogadro number—the resulting ‘solutions’ are no more than the dilution fluid itself. However, numerous physiological and clinical tests have demonstrated for decades, since Hahnemann himself, that this is not the case. Our research objective has been to try to demonstrate that the high dilutions are physically different from the diluent and have, indeed, an ‘individual personality’.

Method

Since any investigation is always difficult in an highly dynamic system we assumed that, should some specific ‘patterns’ exist in the liquid dilution they might be fixed when it is frozen giving rise to specific defects in the crystal lattice of ice, which could be investigated by appropriate means.

To perform this type of studies we selected low-temperature thermoluminescence. This technique, which is well known for archaeological and geological dating,⁴ has been adapted by us to low temperatures⁵ and described in detail in previous publications.^{[6] and [7]} I will here only summarize here its main features.

A 1 cc sample of the dilution under investigation is placed in an aluminum cup and frozen down to liquid nitrogen temperature (−196°C=77°K) following a well defined multi-step process. The frozen 1 mm thick ice disk is then ‘activated’ by radiation (Gamma rays, X-rays or electron beams) which displace electrons from their quantum ground states. The sample is then rewarmed at constant rate (3°C/min) from 77°K to melting point. During that process the electrons, powered by ‘thermal activation’ leave their respective traps and recombine with the empty quantum ‘holes’ releasing their ‘activation energy’ in the form of light as they do so. This light is the thermoluminescent glow that we record.

The analysis of the emitted light shows two main peaks around 120 and 166°K for deuterium oxide and 115 and 162°K for H₂O.⁵ Their relative intensity and shape vary both with the radiation dose and also with the nature of the radiant beam. In particular peak 2 displays a complex structure which can be resolved in a set of individual components by a deconvolution technique.^{[8] and [9]} It is assumed that the ‘defects’ present in the ice crystalline lattice are active luminescent centers, hence that thermoluminescence might be an appropriate tool to study the ‘image’ of the initial liquid samples.

Results

Thermoluminescence is known to be a very sensitive technique and has been used to identify trace compounds. For example see Figure 1, the thermoluminescence emissions of very dilute alumina colloidal sols which show major differences between the 10⁻⁸ g/ml, 10⁻⁹ and 10⁻¹⁰ g/ml solutions.

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Figure 1. Thermoluminescence glow of colloidal sols of alumina irradiated by gamma rays (10 kGy) at liquid nitrogen temperature (77°K).

For homeopathic high dilutions we use deuterium oxide (D₂O, heavy water) as the solute since the signal is 50 times more intense than that of H₂O, due to the more rigid nature of the two ‘arms’ of the molecule. As diluted substances we selected two ionic compounds: sodium chloride (NaCl) and lithium chloride (LiCl). The latter was selected because, like urea and ethanol, it is known to impact on and suppress the hydrogen bonds¹⁰ which are thought to be involved into the high temperature peak (ca 166°K) of the thermoluminescence glow.⁶ Figure 2 shows that the curves recorded for successive dilutions of LiCl (3c, 5c, 7c, 9c) prepared by the classical Hahnemannian method and following the French Homeopathic Pharmacopoeia (150 strokes of 2 cm amplitude in 7.5 s, delivered by mechanical succussion machine) are substantially different.

Display Full Size version of this image (83K)

Figure 2. Thermoluminescence glow of successive dilutions (3c, 5c, 7c, 9c) of lithium chloride in D₂O irradiated by a 2.2 Mev electron beam (6 kGy) at 77°K.

Subsequently, since it appeared that we had a reliable tool for assessing the dilutions we applied the same method to ultra-high dilutions beyond Avogadro’s number.⁶ Figure 3 gives the results and shows evidence that the ‘signature’ peak of LiCl 15c is substantially lower than that of NaCl 15c and lower than succussed pure D₂O. This demonstrates that: ultra-high dilutions are different from their dilution fluid.

Display Full Size version of this image (67K)

Figure 3. Thermoluminescence glow of ultra-high dilutions (15c) in D₂O of LiCl, NaCl and of pure D₂O, diluted and succussed to 15c irradiated by gamma rays (19 kGy) at 77°K.

The high temperature components of the glow (ca 166°K) is linked to the hydrogen bond network. These results have been recently confirmed by another research group.¹¹

In recent and still unpublished experiments we found the same type of ‘scaling’ between increasing dilutions of other compounds, among which potassium dichromate looks particularly interesting.¹²

New prospects

As I said above, in the homeopathic preparation scheme, succussion is an important component of the preparation process of homeopathic medicines, releasing considerable energy in the fluid. In view of this I became interested in recent research on the role of ‘nanobubbles’ in water.¹³ Part of the ‘message’ transferred from one dilution step to the next one might be linked to the nanobubbles created into the liquid by the successive strong mechanical agitation which creates turbulence.

To investigate this, we built special equipment to perform dynamization in gas atmosphere or vacuum. We dynamize the dilution at room temperature (20°C) under a moderate vacuum (2337 Pa=24 mbar) which corresponds to the saturated water vapour pressure at 20°C. Time to reach vacuum is approximately 20 seconds. Dynamization is 150 strokes in 7.5 sec followed by stabilization under reduced pressure for 3 minute. The vacuum is broken reverting to atmospheric pressure in 20 seconds. Figure 4 gives preliminary results which show that the gas-phase seems to play a major role in the ‘personalization’ of the dilutions. Bearing in mind that the number of nanobubbles created into the fluid is of the order of billions (which represents a very large ‘contact’ surface with the surrounding liquid) and that, due to their size, they may remain stable and undisturbed in the dilution for months or even much longer, this might open some new perspectives on our understanding of the homeopathic preparation process.

Display Full Size version of this image (84K)

Figure 4. Thermoluminescence glow of LiCl 15c in D₂O dynamized in a vacuum and in a pure O₂ at 15 bars pressure and irradiated by a 2.2 Mev electron beam (6 kGy) at 77°K. We dynamize the dilution at room temperature (20°C) under moderate vacuum (2337 Pa=24 mbar) which corresponds to the saturated water vapour pressure at 20°C. The time to reach vacuum is approximately 20 seconds, we use our standard dynamization: 150 strokes of approximately 2 cm amplitude in 7.5 sec, followed by stabilization under reduced pressure for 3 minutes. The vacuum is then broken, reverting to atmospheric pressure in approximately 20 seconds.

Acknowledgments

The author thanks Laboratoires BOIRON and the AREVA Nuclear Center of Marcoule for their interest and support.

References

1 J. Teixeira, Can water possibly have a memory? A sceptical view, Homeopathy 96 (2007), pp. 158–162. SummaryPlus | Full Text + Links | PDF (366 K)

2 R. Roy, W.A. Tiller, I. Bell and M.R. Hoover, The structure of liquid water; novel insights from material research; potential relevance to homeopathy, Mater Res Innovations 9 (2005), pp. 93–124.

3 J. Teixeira, A. Luzar and S. Longeville, Dynamic of hydrogen bonds: how to probe their role in unusual properties of liquid water, J Phys Condens Matter 18 (2006), pp. S2353–S52362.

4 Gartia RK. Thermoluminescent materials: past, present and future. In: Sarma HNK, Sumitra P, Basantakumar Sharma H, (eds). Proceedings of Regional Conference on Materials and their Applications (RCMA), February 18–19, 2005, Manipur University, Imphal, India, 2005, p 33–40.

5 L. Rey, Thermoluminescence de la Glace, CR Physi I (2000), pp. 107–110.

6 L. Rey, Thermoluminescence of ultra-high dilutions of lithium chloride and sodium chloride, Physica A 323 (2003), pp. 67–74. SummaryPlus | Full Text + Links | PDF (306 K) | View Record in Scopus | Cited By in Scopus

7 L. Rey, Thermoluminescence of deuterated amorphous and crystalline ices, Rad Phys Chem 72 (2005), pp. 587–594. SummaryPlus | Full Text + Links | PDF (467 K) | View Record in Scopus | Cited By in Scopus

8 B.A. Sharma, Th. Basanta Sing and R.K. Gartia, Critical evaluation of goodness of fit of computerised glow curve deconvolution, Indian J Pure Appl Phys 42 (2004), pp. 492–497.

9 Rey L, Gartia RK, Belon P. Trap Spectroscopic Characterization of D₂O ice and its potentialities in homeopathy. In: Selvasekarapandian S, Murthy KVR, Natarajan V, Malathi J, Brahmanandhan GM, Khanna D, (eds). Macmillan Advanced Research Series. Proceedings of the National Conference on Luminescence and Its Applications (NCLA, 2007) January 18–20, Bharathiar University, India. New Delhi: Macmillan India Ltd., 2007, p 12–17.

10 Ourisson G. Personal communication, 2000.

11 R. van Wijk, S. Basman and E. van Wijk, Thermoluminescence in ultra-high dilution research, J Alternative Complementary Med 12 (2006), pp. 437–443. View Record in Scopus | Cited By in Scopus

12 Rey L, Muchitsch I. Recent unpublished results, 2007.

13 Ph. Vallée, J. Lafait, L. Legrand, P. Mentré, M-O. Monod and Y. Thomas, Effects of pulsed low-frequency electromagnetic fields on water characterized by light scattering techniques: role of bubbles, Langmuir 21 (6) (2005), pp. 2293–2299. Full Text via CrossRef | View Record in Scopus | Cited By in Scopus

Corresponding author. Louis Rey, Chemin de Verdonnet 2, CH-1010 Lausanne, Switzerland.

Homeopathy
Volume 96, Issue 3, July 2007, Pages 170-174
The Memory of Water

This is part of the Homeopathy journal club project described here:

www.badscience.net/?p=490

doi:10.1016/j.homp.2007.03.009
Copyright © 2007 Elsevier Ltd All rights reserved. The defining role of structure (including epitaxy) in the plausibility of homeopathy

Manju Lata Rao^{1, ,} , Rustum Roy^{1, 5}, Iris R. Bell^{2, 3, 4, 5, 6} and Richard Hoover^1
1The Materials Research Institute, The Pennsylvania State University, University Park, PA, USA
²Department of Family and Community Medicine, The University of Arizona, College of Medicine, Tucson, AZ, USA
³Department of Psychiatry, The University of Arizona, College of Medicine, Tucson, AZ, USA
⁴Department of Psychology, The University of Arizona, Tucson, AZ, USA
⁵Department of Medicine (Program in Integrative Medicine), The University of Arizona, College of Medicine, Tucson, AZ, USA
⁶College of Public Health, The University of Arizona, Tucson, AZ, USA
Received 20 March 2007; accepted 27 March 2007. Available online 31 July 2007.

Abstract

The key stumbling block to serious consideration of homeopathy is the presumed “implausibility” of biological activity for homeopathic medicines in which the source material is diluted past Avogadro’s number of molecules. Such an argument relies heavily on the assumptions of elementary chemistry (and biochemistry), in which the material composition of a solution, (dilution factors and ligand–receptor interactions), is the essential consideration.

In contrast, materials science focuses on the three-dimensional complex network structure of the condensed phase of water itself, rather than the original solute molecules. The nanoheterogenous structure of water can be determined by interactive phenomena such as epitaxy (the transmission of structural information from the surface of one material to another without the transfer of any matter), temperature–pressure processes during succussion, and formation of colloidal nanobubbles containing gaseous inclusions of oxygen, nitrogen, carbon dioxide, and possibly the remedy source material.

Preliminary data obtained using Raman and Ultra-Violet–Visible (UV–VIS) spectroscopy illustrate the ability to distinguish two different homeopathic medicines (Nux vomica and Natrum muriaticum) from one another and to differentiate, within a given medicine, the 6c, 12c, and 30c potencies. Materials science concepts and experimental tools offer a new approach to contemporary science, for making significant advances in the basic science studies of homeopathic medicines.

Keywords: homeopathy; succussion; materials science; structure of water; epitaxy; nanobubbles

Article Outline

Introduction

Overview

Materials Science Models for homeopathic medicine

Implications of materials science models for basic science research methods in homeopathy

Preliminary studies of homeopathic medicines using Raman and infrared spectroscopy

Method

Results

Conclusions

Acknowledgements

References

Introduction

Overview

The key stumbling block to serious consideration of homeopathy is the alleged “implausibility” of biological activity for homeopathic medicines in which the source material is diluted past Avogadro’s number of molecules (6×10²³), because the remedy must be identical to the solvent. Negative studies of homeopathy are self-evidently correct from the skeptics’ perspective, because of this error.¹ The implausibility argument leads skeptics to ignore or reject positive evidence from numerous basic science, preclinical, and clinical studies showing effects of homeopathic medicines different from controls, in vitro and in living systems.² On the other hand, proponents predictably reject the negative and focus on positive studies, often uncertain how to address the black box nature of homeopathic medicines. Both skeptics and proponents of homeopathy have generally overlooked a large body of literature in the materials science field that could help resolve this impasse with systematic data.³

Thoroughly, established materials science concepts and research data render the implausibility hypothesis for homeopathy irrelevant. One example suffices. Diamond is the hardest material in nature and graphite among the softest. Yet they can be inter-converted with zero change of composition in microseconds.

The available studies enable significant hypothesis-driven advances in the rigorous study of the nature of homeopathic medicines. The purpose of this paper is to outline the key aspects of materials science considerations in developing experimental models for understanding homeopathic medicines and to summarize preliminary findings from hypothesis-driven studies in our laboratory on clinically known polychrests such as Nux vomica (Nux vom) and Natrum muriaticum (Nat mur).

Materials Science Models for homeopathic medicine

Chemists and medical scientists largely continue to focus reductionistically on the presence or absence of specific molecular species present in water vapor or liquid water without consideration of the ways in which these species are organized in space. From a chemical perspective, the dilution aspects of remedy preparation are the key issue, because of a lack of source molecules for potencies at or beyond 12c or 24c× (10⁻²⁴ dilution). Even when chemists focus on water itself, they emphasize the fleeting stability of hydrogen bonding between given water molecules,⁴ rather than the larger complex structural formations of water or the weaker forces that may favor formation of stable oligomeric and polymeric structures, involving the collective organization of many different water molecules within the condensed liquid phase.

In contrast, materials scientists focus on the organizational network arrangement of the water structures in three-dimensional (3-D) space. In a recent paper, Roy et al.³ presented the detailed technical aspects of the materials science argument concerning ultradilute sols including homeopathic medicines at length. For materials scientists, the succussion aspects of remedy preparation are the key consideration. Temperature and pressure can modify such water structures, leading to nanoheterogeneity of larger structures of water molecule “clusters” within liquid water. Succussion introduces intense turbulence and changes in pressure in any solution,⁵ as well as leading to the formation of nanobubbles in solution.

In brief, the plausibility argument for homeopathy is that liquid water, the primary solvent for source materials in which homeopathic medicines are made, is itself an anomalous substance and has many very different structures. As part of the natural nanoheterogeneity of water structure per se (as contrasted with its composition or the presence of solute molecules), processes such as epitaxy, pressure changes during succussion, formation of colloidal nanobubbles containing gaseous inclusions of oxygen, nitrogen, carbon dioxide, and possibly the remedy source material, and electromagnetic field effects play a role in altering water structure. Previous work by Elia and Niccoli⁶ and Rey,⁷ using different technical methods, respectively, to release heat or light from homeopathic medicines in potency, point to the ability to disrupt what appears to be order or structure in remedy solutions as compared with remedy-free control solvents.

In terms of nanoheterogeneity, water can take on many possible oligomeric and polymeric structures, ie, form complex networks of water molecules in 3-D space, held together by various forces that include not only hydrogen bonds (relatively strong), but also van der Waals forces (much weaker). Even if specific molecules or small molecular complexes leave their places in the network, other water structure complexes can take their places within the network structure itself, thereby maintaining the overall nanostructures within the solution, in part via configurational entropy or electromagnetic forces maintaining organizational stability of the network.⁸

Notably, research in the field of complex systems and network science has shown that, within a highly complex network, loss or disruption of a given member or node, which is a point of interconnection with other members of the network (eg. a water molecule or small complex of water molecules) does not destroy or significantly disrupt the overall network organization.^{[9] and [10]} With complexity in liquid water as a whole comes the capacity for overall stability that is not possible in the simpler organizational structures of water on which chemists usually focus.

Epitaxy is the transfer of information, not material, from the surface of one material, usually solid, to another, usually liquid¹¹. The substrate (eg. remedy source material) acts as a seed crystal for the formation of the structure in the recipient surface material (eg. network organization of water structures). Semi-conductor manufacturing often utilizes epitaxial growth to generate specific types of microtransistors and integrated circuitry. In addition to the original source material that uniquely contributes to remedy preparation, deliberate additives in homeopathic medicines, such as ethanol, and/or possible contaminants from succussion, such as silicates from glass container walls, may also stabilize the water molecule structures with their own epitaxial capabilities. Thus, epitaxy can interact with temperature–pressure factors to create unique patterns of information without the transfer of material.

In terms of “seeding” formation of informational structures within water, initial empirical observations on homeopathic medicines suggest that the passage of time between the original remedy preparation and the testing procedures can alter experimental findings. In calorimetric and thermoluminescence studies on homeopathic medicines, the time factor contributes to differences in the magnitude and even the direction of the divergence between remedy and control solutions.^{[4] and [12]} Overall, the behavior of homeopathic medicine liquids in terms of their structural properties in the basic science literature exhibits a somewhat unpredictable, self-organizing quality.

As additional data emerge, these lines of research may facilitate advances in understanding the nature and mechanisms of variability in clinical responsivity to homeopathic medicines.^{[13] and [14]} Water is an hub molecule (a highly interconnected and influential molecule) in most of the biochemical reactions in the body.¹⁵ In a more speculative but testable vein, seeding informational changes in body water at global and local levels¹⁶ of scale could be one way in which homeopathic medicines interface with patients to induce patterns of system-wide and local healing responses.¹³

Implications of materials science models for basic science research methods in homeopathy

Materials science models for the nature of homeopathic medicines leads to more rational selection of specific methodologies for basic science studies. For example, many earlier studies of homeopathic medicines relied on nuclear magnetic resonance (NMR) techniques.^{[17] and [18]} However, NMR spectroscopy provides information on structure of individual atoms in a pure molecule better than on complex networks of molecules. Technically, NMR also requires addition of substances to prepare a liquid for testing. The necessity of adding factors in the process of making observations can introduce unintended contaminants into the measurement process.

In contrast, the light scattering technologies of Raman spectroscopy and Fourier transform (FT) infra-red (IR) spectroscopy permit examination of remedy samples without fixatives or other potential contaminants. Furthermore, Raman and infra-red spectroscopic techniques allow the co-operative nature of structural differences to be detected. Recent studies¹⁹ of microscopic dynamics of hydrogen bonded liquids indicate the existence of highly directional H-bonds, whose energy value normally range between 8 and 25 kJ mol⁻¹ induces different chemical–physical properties and different local environments. As the mean lifetime of H-bonds is in the picosecond timescale, such structures are considered as transient species in dynamic equilibrium.

Our recent work has established the importance of the structure of water on its properties,³ we examined the structures of many water and alcohol-based homeopathic remedies. The results show that such materials can be easily distinguished from the pure solvent, and from each other, by the use of UV–VIS (ultraviolet–visual) and Raman spectroscopy, but Fourier transformed infra red (FTIR) spectroscopy proved insensitive to these differences. This opens up a whole new field of endeavor for inorganic materials scientists interested in developing a scientific basis for the efficacy of homeopathic remedies. The assumption of this study is that the joint employment of the two methodologies: optical spectroscopic tools and electronic microscopic tools can furnish a closer reference picture for the comprehension of the structural changes in the liquid phase besides providing an independent understanding on the role of the ‘active ingredient’ in a homeopathic medicine.

Also we believe that our very preliminary efforts in using cryo-scanning electron microscopy (cryo-SEM) and cryo-transmission electron microscopy (cryo-TEM) may eventually possibly provide definitive evidence of the presence, and the effects, of nanobubbles on homeopathic medicine studies.

Preliminary studies of homeopathic medicines using Raman and infrared spectroscopy

Method

A Food and Drug Administration-regulated homeopathic pharmacy (Hahnemann Laboratories, San Rafel, CA) prepared samples of two different test solutions in 16 ounce (450g), clear glass bottles [Type I borosilicate glass] previously annealed at temperatures between 600–700 °C for 15 minutes. One of the solutions, Nat mur (mineral: Sodium Chloride) and the other Nux vom (plant remedy, purchased as tincture from Boiron) were diluted by the standard Hahnemannian techniques in 95% ethanol and succussed: a 30c potency is diluted (1/100)³⁰ or 10⁻⁶⁰ from the original material. They were hand-succussed by trained experts [http://www.hahnemannlabs.com/preparation.html] 30×20=600 times during the manufacturing process. Each bottle was coded with an unique number, the bottles were shipped together by overnight courier in the same box, with temperature sensor.

We have used UV–VIS, IR, FTIR, and Raman spectroscopy for the bulk “liquid” which in most cases is either water or a mixture of water and ethanol (95% ethanol). UV–VIS spectroscopy and Raman spectroscopy proved to be useful tools to investigate the subtle but significant changes in the structural parameters in both water and alcohol based remedies. (For details refer to 20). While other techniques such as freezing point depression; acoustic loss spectroscopy, ellipsometry, viscosity, surface tension, have been explored and will eventually be used in depth to measure entirely different properties, we report here our experience with the major spectroscopic techniques which are widely available.

(a) UV–VIS spectrophotometer: VARIAN, Model CARY 100, run in dual beam mode,

(b) FTIR spectrophotometer: Thermo Nicolet, Model NEXUS 670, run in attenuated total reflection (ATR) mode, and

(c) Raman spectrophotometer: Inphotonics, Model RS2000-3b-785, using an InPhotonics fiber optic immersion probe.

Results

Nearly 200 runs were made to calibrate every step in the experimental configurations and procedures used for the different instruments. In the dual beam UV–VIS, the many experimental options are all tested separately to ensure that any differences within the data obtained on our samples are well above the instrument noise measured in the calibration run data. The data are obtained largely at different times scales by different individuals gave consistent results. We note that at very low signal levels, instrument noise coupled with artificial computer generated sensitivity can produce data that are not reliable. Hence, we operate the instruments in the sensitivity ranges in which we sacrifice some precision for reproducibility. In the Raman spectrometer, careful attention is paid to the positioning of the probe within the sample container, and stray light is eliminated by turning off all the room lights whenever data are being collected. Details of this work are published elsewhere.²¹

One of the objectives in undertaking this work is to examine evidence which would suggest reliability of physical properties, assuming structural changes in solvents, especially in ultradilute and dilute sols, an excellent example of the class of materials being homeopathic remedies. For our study, we chose to study Natrum muriaticum and Nux vomica, obtained from Hahnemann Laboratories. Both Nat mur and Nux vom are prepared in 95% ethanol. Three types of analyses are presented:

(a) Comparison of specific homeopathic remedies with different potencies [Nat mur 6c, 12c, 30c, and Nux vom 6c, 12c, 30c].

(b) Comparison between two different remedies of the same potency [Nat mur vs Nux vom 6c, 12c, and 30c].

(c) Comparison of the two homeopathic remedies with unsuccussed and succussed plain ethanol.

Figure 1 shows a comparison of Nux vom and Nat mur, 6c, 12c and 30c, showing representative UV-spectra demonstrating the differences between the remedies. In Figure 2 (a), and (b) we show the envelope of differences within a series of 10 preparations of each remedy of Nat mur and Nux vom. The spectra show clear differences in the same potency of an individual remedy for both Nat mur and Nux vom.

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Figure 1. Comparison of two different homeopathic medicines: Natrum muriaticum (NM) and Nux vomica (NV) showing representative UV-spectra demonstrating the differences between the remedies.

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Figure 2. Envelope of differences within a series of 10 preparations supplied of each Homeopathic medicine: Nat mur and Nux vom.

A comparison was also made between the unsuccussed ethanol and the Nat mur and Nux vom samples as shown in Figure 3. The Roy et al paper³, on “structure of water” clearly evidence the role of succussion besides epitaxy and other temperature effects, on the structure of liquids. Under the “normal” succussing procedures, it can be argued that very considerable pressures (of the order of 10 kbar) could be generated as a result of the shaking. Dachille and Roy²² showed that mere grinding in a mortar and pestle gives rise to high pressures up to 20 kbar, and the figures for force per unit area are strongly dependent on the size of the water particles and the velocity of the shaking. By analogy with similar liquids, such as ethanol, there will be many different structures of water formed both by the pressures generated in succussing in some combination with the epitaxy on any additives.

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Figure 3. UV–VIS spectra of: (a) succussed and unsuccussed ethanol, (b) comparative UV–VIS spectra of Nux vom (NV) 6c, 12c, 30c with unsuccussed ethanol, (c) comparative UV–VIS spectra of Nat mur (NM) 6c, 12c, 30c with unsuccussed ethanol.

It may be noted from Figure 3 that the absorption spectra for unsuccussed ethanol is significantly different from: (a) the succussed ethanol and (b) succussed homeopathic remedies, Nat mur and Nux vom. The difference may be attributed to the variation in intra and inter-molecular association of ethanol and water and the generation of both transient and stable nanobubbles. The work of Tyrrell and Attard at Australian National University has proved beyond any doubt that nanobubbles do exist and persist.²³ FTIR Spectra (not shown here) from all the samples of Nat mur and Nux vom overlap neatly, clearly signifying that FTIR is not the most sensitive technique for analyzing the subtle structural differences in these types of samples.

Comparison of homeopathic remedies with different potencies using Raman spectroscopy is done on the two sets of homeopathic remedies: Nat mur and Nux vom. From the spectra shown in Figure 4, a clear distinction in the Raman active modes is noted between the two different remedies as well as among the different potencies of the same remedy. A clear distinction is shown in the spectral peaks from the different potencies, peak positions identified as (a), (b), (c), (d) and (e) in the Raman spectra of Nat mur samples show significant structural changes. While the existence of distinct structural changes in Nat mur and Nux vom remedies is clear from the Raman spectra, significant structural changes are also noted in the spectra of Nux vom between the different potencies, 6, 12 and 30c, peak positions are identified as (a), (b), (c), and (d) in Figure 4b. Further, since all the homeopathic medicines were prepared in 95% ethanol, we analyzed the Raman spectra of unsuccussed and succussed ethanol shown in Figure 5. Note that 6c potency of the succussed ethanol show distinct structural variations.

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Figure 4. Comparison of the Raman spectra of the same potencies, 6c, 12c and 30c, for two different homeopathic medicines. The differences in the peaks identified as (a)–(e) is clearly visible in 30c samples of Nat mur and Nux vom, compared to other diluting of the same medicine.

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Figure 5. Raman spectra of plain ethanol and succussed 6c, 12c, 30c. Note that peak positions identified from (a)–(f) are prominent only in 6c sample. Also note that the intensity of peaks in the unsuccussed ethanol is significantly lower than the succussed samples.

Conclusions

Materials science provides a conceptual and empirical foundation for future research on the nature of all the dilute sols including homeopathic medicines in the physical plane. Processes such as epitaxy, temperature-pressure induced changes in water structure, and nanobubble formation offer testable hypotheses for understanding homeopathic medicines. Although hypotheses regarding seemingly unmeasurable “subtle energies”²⁴ and/or macro-entanglement phenomena^{[25] and [26]} may help explain the fuller nature of homeopathic medicines, the available evidence also suggests that homeopathic medicines can exhibit qualitatively and quantitatively different structural properties from those of unsuccussed or succussed solvents. Even in the case of subtle energies, initial findings indicate the possibility of measuring changes in liquid structure properties from the materials science perspective.^{[27] and [28]}

The convergence of data from different experimental models suggests that it is feasible to study the nature of homeopathic medicines using available basic science tools, notably here, Raman spectroscopy and ultraviolet–visual absorption (UV–VIS) spectroscopy. Reproducibility of findings is feasible within the same Raman equipment, but, not across different Raman spectrophotometers from the same manufacturer at different geographic locations, even for materials other than homeopathic medicines. Fourier transform infrared (FT-IR) spectroscopy cannot differentiate different homeopathic medicines or different potencies of the same remedy from one another. Transmission and structural electron microscopy are promising options for testing the nanobubble hypothesis.

Finally, the materials science perspective provides a possible translational bridge from the emerging complex systems/network science models for clinical responses to homeopathic treatment^{[5], [12], [13], [29], [30], [31] and [32]} to another level of organizational scale, ie, the network structure of the homeopathic medicines themselves. Given the holistic quality of clinical diagnosis and remedy selection in homeopathy, the articulation of holistic (complex network) rather than reductionistic models for both the clinical healing process and the nature of homeopathic medicines is heuristically appealing.

Acknowledgments

The authors gratefully acknowledge financial support for their research from grants from The Council for Homeopathic Research and Education, Inc.; the Friends of Health Foundation; and NIH/NCCAM K24 AT000057.

Conflicts of interests

Dr Bell serves as a consultant to Standard Homeopathic Company/Hyland’s Inc., which did not provide any direct financial support for the research discussed in this paper.

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