Evidence from the Use of Solvatochromic Dyes Indicates that Bulk Pure Water does not Potentise

Abstract

Background Whilst it is assumed that water plays a crucial role in homeopathy, it is unclear what specific functions water has in the generation, propagation and maintenance of homeopathic potencies. Solvatochromic dyes have demonstrated themselves to be reliable indicators of the presence of potencies and so are well placed to begin to address these questions.

Objective The aim of the current study was to ascertain whether pure water could be potentised. If pure water can be potentised, this would indicate that water plays a crucial role in the generation of homeopathic potencies. If it cannot be potentised, then this result points to water being an inert carrier and medium for the potentisation of substances placed in it.

Methods A specially designed potentiser capable of consistently and reliably producing homeopathic potencies to rigorous standards (Pinkus Potentiser) was used to run up potencies of “Water 10M” and Arsenicum 10M in two interchangeable stainless-steel vials. “Water 10M” and Arsenicum 10M, along with unsuccussed control water, were then tested against solutions of the solvatochromic dye phenol blue by following the absorbance of the dye at 658 nm.

Results “Water 10M” gave results no different statistically from unsuccussed water (p > 0.05) with no change over the time course of the experiments, whereas Arsenicum 10M gave statistically different results (p < 0.001) with increasing divergence from both “Water 10M” and unsuccussed water over time. Similar results were obtained when the vials in which Arsenicum 10M and “Water 10M” were produced were exchanged, so that the vial in which Arsenicum 10M had been produced was then used to produce “Water 10M” and vice versa, indicating the results could not be explained by any anomalies in the production vials themselves.

Conclusion Results using the solvatochromic dye phenol blue to detect and measure homeopathic potencies indicate that bulk water itself does not potentise and that unsuccussed water and water that has been succussed are no different. This indicates that what is potentised during cycles of dilution and succussion are the substances placed in water and not the water itself. In turn, this indicates that water is simply a carrier for the homeopathic signal. Evidence from the practice of homeopathy supports this conclusion.

Keywords

Pinkus potentiser - water - solvatochromic dyes

Introduction

One of the central questions in homeopathy is what role water plays in the generation, propagation and maintenance of the homeopathic signal. There appears to be an unquestioned assumption that water's role is crucial and that without water there would be no homeopathy. However, direct evidence for this assumption is lacking, and in fact, there is some evidence to suggest that we would be wise to question the role of water (or lack of) more closely.

For instance, homeopathic medicating potencies are kept for long periods by pharmacies and practitioners in 90-96% ethanol.[1] Any hypothesis involving complex arrangements of water hydrogen bonding requires increasingly convoluted and questionable arguments to support a ‘memory of water’ hypothesis in the presence of such large and disruptive volumes of ethanol.

In addition, most homeopathic prescriptions involve lactose/sucrose tablets to which a few drops of medicating potency have been added and these tablets are very often then dried. These tablets will contain solely water of hydration, or around one water molecule per lactose or sucrose molecule. Such bound water will have little freedom of movement and certainly no opportunity to form any three-dimensional structure with other water molecules.

A third argument against water playing a crucial role in homeopathy is the practice of giving remedies by olfaction. This method of administering remedies has been practised by generations of homeopaths from Hahnemann, Hering and Clarke to the present day and is well documented.[2] Aside from arguments as to the role of the olfactory epithelium and the vomeronasal organ in remedy transduction,[3] it is known that volatility requires a molecular mass of less than 300 Daltons.[4] As the molecular mass of water is 16 Daltons, this requires that all the information for all remedies at all potencies must, at best, be encoded by 20 water molecules and that those 20 water molecules must remain in unique three-dimensional arrangements during their travel from the opened bottle of tablets/liquid to engage with the patient's olfactory epithelium—a distance of approximately 3 to 5 cm, without any disruption to that unique arrangement occurring. It is hard to formulate any mechanism within the known properties of water that would allow for this possibility.

If water lacks any sufficient three-dimensional structure in 90-96% ethanol, on lactose/sucrose tablets or in its vapor phase, to account for the maintenance of the homeopathic signal, what role might water play? A first step in answering this question would be to see if pure water itself can be potentised. If the answer is yes, and an effect is seen with a reliable test system, then this would indicate that water is capable of generating a signal through succussion, even if water is not necessary for the maintenance of the signal. If the answer is no, and no effect is seen with a reliable test system beyond that of control, then water can neither be the source of the homeopathic signal nor is it necessary for the maintenance of the signal. This would then relegate water to an inert carrier of the signal, along with other carriers such as lactose (see Discussion).

The study reported here seeks to answer the specific question as to whether bulk pure water can be potentised using a solvatochromic dye detection system involving phenol blue. Previous studies have indicated that succussed water and unsuccussed control water have no effect on solvatochromic dyes BDN, ET33 and MV compared with potencies of Antimonium crudum,[5] on the dye MV compared with Phosphorus 30C,[6] on ET33 compared with potencies of Mercurius corrosivus 30C[7] and on the dyes Coumarin 7 and ET33 compared with potencies of Glyphosate.[8] However, a more rigorous study is required to try to establish unambiguously whether succussed and unsuccussed water are different in relation to their interaction with solvatochromic dyes, and consequently, whether it could be concluded that bulk pure water can be potentised or conversely whether it is solely an inert carrier of potencies.

Carrying out a comparison of succussed water and of a known substance that has gone through a process of successive dilution and succussion steps faces several substantial challenges. The succussion process must, as far as possible, be the same for both pure water (in the current study to a theoretical level of 10M) and water with an assigned substance present (in the current study Arsenicum to a level of 10M) and be performed at the same time, with water from the same source. In addition, assays must be subject to stringent conditions if the question central to this study is to be adequately answered. This has been achieved by using a purpose-built potentiser of precise specifications (Pinkus Potentiser) together with using a rigorous set of methods throughout the production, transport and testing procedure (see materials and methods).

Materials and Methods

Potentiser

Arsenicum 10M and “Water 10M” were produced using the Pinkus Potentiser ([Fig. 1]), which is based on a combination of the Korsakovian and fluxion principles of potentisation.[9] This potentiser was designed and introduced to mark a high industry standard for potentisers and as such all parts are designed to be robust and replaceable. In addition, specifications include the addition of calibration devices to ensure reproducibility in function. The machine has two stainless steel potentising vials (A and B) both of which are designed to be removed for autoclaving between remedies as a line clearance procedure. The vials are made from 304 stainless steel, which is highly resistant to the solvating action of pure water and, unlike glass, does not shed nanoparticles of silica into solution. An accelerometer has been used to calibrate and measure the downforce of succussion and fits in the space of an empty vial. The accelerometer is a pressure transducer and when connected to a computer with appropriate oscilloscope software, it converts the deceleration signal into a voltage peak representing a downforce in ‘g’ (the average acceleration produced by gravity at the Earth's surface, which is 9.806 m/s²)

Fig. 1 Photograph of the Pinkus Potentiser (above) and of the two vials in which potencies are made (below left) and the vials in situ (below right).

The Pinkus Potentiser is designed and calibrated to produce a force of 3 g on impact with each succussion stroke. Although it has a variable succussion number setting, the machine is set to give 30 succussions per dilution cycle. Each cycle lasts 16.5 seconds, thereby producing 218 cycles per hour. 218 cycles per hour produces a 10M in approximately 45 hours if running continuously. In practice, the potentiser is run only during working hours, resulting in a running time of approximately 3 weeks.

The succussion procedure itself is mechanically affected by a servo motor programmed to control an oscillating beam at the end of which is a harness for the two identical heart-shaped vials placed side by side. These sit above a pair of neoprene pads, and the conical base of each vial is arranged to strike the pad eccentrically to allow wear to be managed. The tensile strength of the pads is checked and maintained at 85 International Rubber Hardness Units. The rise and fall of the vials onto the pads constitute the succussion strokes necessary to produce potencies. The twin vials are designed in the form of heart-shaped chambers, each with three openings to allow the input and exit of water and air. One opening is for water in, the second for water out, and the third for blasts of filtered compressed air from the onboard oil-free compressor. An onboard programmable logic controller controls the numerous steps in each potency cycle. Hence, the 1:100 dilution is affected by sequential blasts of air to efficiently empty the cell in the least time. Water is delivered by two powerful computer-controlled water pumps set to deliver at the rate of 37 L/h at 40 psi. The pressurised air used to empty cells of water is filtered and also set at 40 psi. The time taken to withdraw water, air blast and refill for each cycle, as stated above, lasts a total of 16.5 seconds.

The water used in the potentiser is purified using a 5 micron filter followed by reverse osmosis (18 MΩcm conductivity) into a 40 L bulk tank, recirculated via a resin cartridge and UV light. The Pinkus Potentiser is designed to run two samples simultaneously and uses 300 L of purified water for the 10 M potency, which is produced from approximately 1,000 L of mains water. The two vials are run with the same batch of ultrapure water, at the same time, with identical downforces at each and every succussion.

For the present study, vials were baked for 1 hour at 200°C before reuse. This allows for the production of Arsenicum 10M and “Water 10M” in vials A and B respectively and then a subsequent production run of Arsenicum 10M and “Water 10M” in vials B and A respectively to ensure any anomalies with one of the vials would be cancelled out.

In addition, a mu-metal shield has been installed between vials A and B. In this way, two remedies (or in this study Arsenicum and water) can be run simultaneously without the concern of potential transfer of signal between vials through field effects.[10]

Assays

Post-production, Arsenicum 10M and “Water 10M” have been kept at a distance of at least 30 cm from each other at all times. Following the final succussion step in the Pinkus potentiser, potencies were transfered to amber glass bottles from the same batch and ethanol (Kimia, UK) added to 96% v/v. The bottles were wrapped in foil and sent from Ainsworths production centre in Surrey, United Kingdom, to the testing laboratory in Oxford under separate cover to minimise any contamination by potential field effects.

Unsuccussed control water was also made up to 96% ethanol in the same amber glass bottles as the “Water 10M” and Arsenicum 10M. Control water, “Water 10M” and Arsenicum 10M were, therefore, as far as possible, materially equivalent.

Assays were conducted as soon as samples were received and over a 24-hour period or less. This was to ensure that environmental conditions varied as little as possible over the time taken to perform at least five assays each of unsuccussed water, “Water 10M” and Arsenicum 10M. Rapid assaying also ensured that any potential field effects between unsuccussed water/ “Water 10M” and any extraneous potencies, even at a distance, would be kept to a minimum.

Upon arrival at the Oxford laboratory both Arsenicum 10M and “Water 10M” were tested, along with unsuccussed control water, using the following test procedure. The assay method was kept as simple as possible to avoid any unnecessary manipulations which might introduce unwanted variations between assays. Phenol blue (N,N-dimethyl-indoaniline) was made up in dimethyl sulfoxide to a concentration of 1 mM the previous day. For each assay 50 µL of unsuccussed water, “Water 10M” or Arsenicum 10M straight from the amber glass bottles described above were added to 2.95 mL of double distilled water with a conductivity of 0.25 µScm⁻¹ in a Brand UV cuvette. Immediately following this 2 µL of phenol blue solution was added and the cuvette inverted three times and the absorbance at 658 nm was noted. Successive readings at the same wavelength were made over a period of minutes/hours, with the cuvette returned to a black film canister between measurements. Absorbances over time were then plotted out for unsuccussed controls, “Water 10M” and Arsenicum 10M. Due to the time required to clean the potentiser vials and run up potencies, a period of 3 weeks elapsed between the first set of assays (Arsenicum 10M from vial A/ “Water 10M” from vial B) and the second set of assays (Arsenicum 10M from vial B/ “Water 10M” from vial A) (see Discussion).

Assays were performed in double distilled water, rather than buffered solution. This was done to keep the number of system components and manipulations to a minimum. Preliminary assays with control solutions from different amber glass bottles showed that buffering was not necessary and that good stability was maintained.

Statistical Analyses

Statistical analyses were performed using Excel 2019. Error bars are based on 3 standard deviations (3σ), meaning 99.7% of values fall within the error bars. This was deemed to be a more cautious representation of the data than at 1σ or 2σ.

Instrumentation

Assays were performed using a Shimadzu UV2600 UV/vis spectrophotometer. Conductivity measurements were made with a DDS-17 conductivity meter.

Results

[Fig. 2] shows the results from the production cycle using Arsenicum 10M from vial A/“Water 10M” from vial B tested against phenol blue. Mean values are unsuccussed (control) water 0.134, “Water 10M” 0.1335 and Arsenicum 10M 0.1307 at time = 0, diverging to control water 0.133, “Water 10M” 0.1326 and Arsenicum 10M 0.124 at time = 180 min. “Water 10M” gave results no different statistically from unsuccussed (control) water (p > 0.05) with no change over the time course of the experiments, whereas Arsenicum 10M gave statistically different results (p < 0.001) with increasing divergence from both “Water 10M” and unsuccussed water over time.

Fig. 2 Change in absorbance of phenol blue at 658 nm over time with control added (n = 10) (upper trace), ‘Water 10M’ (n = 7) (middle trace) and Arsenicum 10M (n = 6) (lower trace). Arsenicum 10M made in vial A of the potentiser, ‘Water 10M’ in vial B of the potentiser (see text for details of assays). Final values at t =180 minutes; control 0.133 (SD = 0.0015), ‘Water 10M’ 0.1326 (SD =  0.00175), Arsenicum 10M 0.124 (SD = 0.0018). Error bars are 3σ.

[Fig. 3] shows the results from the production cycle using Arsenicum 10M from vial B/ “Water 10M” from vial A tested against phenol blue. Mean values are unsuccused (control) water 0.121, “Water 10M” 0.12058 and Arsenicum 10M 0.1166 at time = 0, diverging to control water 0.1199, “Water 10M” 0.11918 and Arsenicum 10M 0.1124 at time = 180 min. Again “Water 10M” gave results no different statistically from unsuccussed water (p > 0.05) with no change over the time course of the experiments, whereas Arsenicum 10M gave statistically different results (p < 0.001) with increasing divergence from both “Water 10M” and unsuccussed water over time. In both sets of experiments, the very slightly lower value for “Water 10M” with respect to control water is not statistically significant and is therefore likely coincidental.

Fig. 3 Change in absorbance of phenol blue at 658 nm over time with control added (n = 5) (upper trace), ‘Water 10M’ (n = 5) (middle trace) and Arsenicum 10M (n = 10) (lower trace). Arsenicum 10M made in vial B of the potentiser, ‘Water 10M’ in vial A of the potentiser (see text for details of assays). Error bars are 3σ.

Discussion

Pilot studies to ascertain the best experimental protocol to answer the objective of the current study found that Brookers merocyanine and β-cyclodextrin showed significant differences between Arsenicum 10M versus unsuccussed water and “Water 10M” over the several hours of the experiments, with unsuccussed water and “Water 10M” on the other hand showing minimal differences. However, fluctuations in absorbances over time meant controls were not as stable as demanded by the aims of the current study, so dyes producing more stable control absorbances were sought.

Phenol blue shows minimal fluctuations in absorbance over time and has a relatively stable absorbance compared with other solvatochromic dyes over the time course of these studies. While this produces stable control plots it means that sensitivity is reduced. Nevertheless, several studies have shown that phenol blue responds to potencies over a wide range of conditions[11] [12] [13] and it was chosen for these reasons.

[Figs. 2] and [3] show that differences between “Water 10M” and unsuccussed controls using phenol blue are minimal in both experiments and statistically not significant. Arsenicum 10M, however, does show a statistically significant difference in its effect on phenol blue compared with “Water 10M” and unsuccussed controls. This difference is maintained even on switching the vials in which potentisation is performed. The effect cannot, therefore, be due to any anomalies in one of the vials.

Before drawing conclusions from the above results, however, several caveats and issues need to be discussed.

It will be noted that absolute values of absorbances in [Figs. 2] and [3] are different between the first set of results and the second. This is because the first set of runs (control, ‘Water 10M’ from vial B, Arsenicum 10M from vial A) were performed as quickly as possible over a 24-hour period to avoid any (unknown) environmental variations over time that might affect the samples and/or the dye. The vials were then baked at 200°C for 1 hour and new potencies run up. This took around 3 weeks altogether. It is not desirable to use old dye solution to do further assays so a new dye solution was made up. No two dye solutions will have exactly the same concentration of dye (and hence absorbance) and, in addition, conditions may not be exactly comparable (temperature, humidity, etc.) from one month to the next – hence the difference between absolute absorbances of controls. The second set of assays (control, “Water 10M” from vial A, Arsenicum 10M from vial B) were performed 3 weeks after the first set of assays, and again over 24 hours to avoid any (unknown) environmental variations. Differences between controls, “Water 10M” and Arsenicum 10M are therefore valid for both sets of results, but absolute values cannot be compared between assay sets where the intervening time frame is as long as 3 weeks.

Strictly, potentisation, as defined in homeopathy, consists of coupled dilution and succussion steps. This study has demonstrated that bulk water does not appear to potentise through succussion alone. At every step, more water is added to the potentisation vial; therefore water is never diluted and so it is not possible to state that the final product is Water 10M. Hence, it is referred to as “Water 10M” throughout to emphasise this discrepancy. We have not demonstrated that water cannot be potentised under any conditions, only that bulk water does not appear to be potentiseable. It may in fact be impossible to demonstrate that water can or cannot be potentised in the same way that other substances are potentised. This is because water cannot be diluted effectively. Any solvent that could be used as a diluent would be hygroscopic, and so absorb water from the air at every step. Hence, the concentration of water in the vials would remain relatively constant. For the purposes of the present study, however, we have demonstrated that the water into which substances have been placed appears to play no part in their potentisation.

Another (though unlikely) possibility that must be addressed is that on adding ethanol at the last step in the potentisation process, this may differentially effect “Water 10M” and Arsenicum 10M, possibly rendering the former inactive in some way and so 96% ethanol /”Water 10M” is not the same homeopathically as “Water 10M”, or that both “Water 10M” and Arsenicum 10M are equally affected by adding ethanol. This is an extremely difficult question to answer practically. We have found that microbial growth occurs quite quickly in water samples in the absence of ethanol, giving spurious results even when samples are analysed as soon as they arrive from the pharmacy. We have worked on the basis that ethanol does not differentially or equally affect “Water 10M” and Arsenicum 10M. If it were the case that ethanol affects “Water 10M” and Arsenicum 10M one would also have to entertain the possibility that all remedies and all potencies are differentially or equally affected homeopathically upon addition of ethanol during their production, and there is no evidence that is the case.

It should also perhaps be pointed out that potencies can be produced by trituration alone, and indeed all LM potencies are produced to the 3cH level by this method. Trituration does not involve bulk water at all but relies on lactose as the medium in which potentisation occurs, so the results of the experiments reported here may not come as a surprise.

The simplest conclusion from the results shown in [Figs. 2] and [3], and taking into account the issues raised above, is that bulk water itself does not potentise and that only substances put into water (in this case Arsenicum album) produce potencies. In short, this evidence points to water being an inert carrier of homeopathic potencies and not a necessary component for the generation of potencies.

If water is not necessary for the generation of potencies, beyond it providing a medium for succussion, what then might the role of water be? From the arguments presented earlier and derived from the realities of homeopathic practice (storage in 90–96% ethanol and on lactose/sucrose tablets, as well as the administering of remedies by olfaction), water is not necessary for the maintenance of potencies either. This leaves a possible role for water in the propagation or spread of the homeopathic signal.

Whilst definitive evidence is still lacking to allow confirmation or rejection of this possible role for water, a previous study looking at the spread of Phosphorus 30C through a large lake system in Brazil did provide evidence that the spread of Phosphorus 30C was significantly faster than the flow of water from the point of seeding to the final lake in the series.[6] This suggests that Phosphorus 30C spread or replicated independently of the water in which it was placed.

It seems likely therefore that water is not necessary for the generation, propagation or maintenance of the homeopathic signal and that water is nothing more than an inert carrier for potencies.

It would be interesting in the future to ascertain what other substances can act as carriers of homeopathic potencies and, if so, are there any common features which turn out to be necessary in order for those substances to fulfil that function. Lactose is clearly a strong candidate and Hahnemann originally used fragments of oyster shell as a carrier.[14]

More importantly perhaps is what the results and arguments presented above mean for any hypotheses as to the mechanism of action of homeopathy. If water is not central to homeopathy, what is?

Conclusion

Results from testing Arsenicum 10M, “Water 10M” and unsuccussed control water against the solvatochromic dye phenol blue indicate that bulk water itself does not potentise and that unsuccussed water and water that has gone through many cycles of succussion are no different. On the other hand, Arsenicum 10M, which has been treated identically to “Water 10M” in terms of the number and strength of succussion steps using the Pinkus Potentiser, is significantly different in its effect on the dye phenol blue. This indicates that what is potentised during cycles of dilution and succussion are the substances placed in water and not water itself. Evidence presented from previous studies supports this conclusion, as do arguments derived from the practice of homeopathy in relation to the storage and administering of homeopathic remedies.

Highlights

• The aim of the current study is to compare pure water that has been succussed to a theoretical level of 10M with a standard remedy (Arsenicum album 10M) to the same succussion level using a solvatochromic dye test system.

• The result indicates that water that has been succussed is no different from unsuccussed control water and that during the potentisation process only substances placed in water are potentised, not water itself.

• The implications of this result, together with evidence from the practice of homeopathy, are that water is not involved in the generation, propagation or maintenance of the homeopathic signal, but is simply a carrier for the signal.

Conflict of Interest

None declared. Tony Pinkus holds the position of Superintendent Pharmacist & Technical Director of Ainsworth Homeopathic Pharmacy. He also owns Ainsworth Pharmacy and designed the Pinkus Potentiser. However, all experimental work and assays were executed independently of Ainsworths pharmacy.

Acknowledgements

Financial assistance from Ainsworths Homeopathic Pharmacy is gratefully acknowledged.

• References

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Publikationsverlauf

Eingereicht: 10. Juli 2023

Angenommen: 16. August 2023

Artikel online veröffentlicht:
13. Dezember 2023