27th August 2007.

Dr. Peter Fisher,
Editor, Homeopathy,
The Royal London Homoeopathic Hospital,
60 Great Ormond Street,
London, WC1N 3HR.

Dear Sir,

We wish to draw to your attention serious anomalies and incongruities in the UV absorption data presented in the paper by Rao et al., published in your July 2007 issue [1].

In a study of this nature, which in effect is examining multiple samples of ethanol, the over-riding concern must be absolute uniformity in the source of the solvent. For the data to be valid, it is essential that every drop of ethanol used must be sourced from the same stock bottle. However, the authors fail to make any mention of this point, and it is clear from the results presented that the source of ethanol in this investigation was most certainly not uniform.

The most striking anomaly is the UV spectrum presented for “plain ethanol”, a single trace repeated three times in figure 3. The provenance of this sample is not recorded. This trace reveals extremely high absorbance (greater than 0.8 absorbance units) at 250nm, falling off steeply towards 400nm but still above 0.4 units by 350nm, and demonstrating an absorbance peak of 0.65 units with a l-max of about 330nm. It is simply impossible to represent this trace as being ethanol of any recognised degree of purity. Spectroscopic grade ethanol has an absorbance of less than 0.05 units between 250 and 400nm [2], and even USP/NF pharmaceutical grade ethanol has an absorbance of less than 0.3 units at 250nm, falling off to less than 0.1 units by 270nm [3]. If the substance measured by the authors as “plain ethanol” was indeed ethanol at all, it is clear that it contained extremely high levels of impurities, possibly including acetone.

In contrast, the spectra of the samples which were diluted and succussed (Nat mur, Nux vomica and the “succussed ethanol” with no mother tincture), and which were presumably all supplied by Hahnemann Laboratories as detailed on page 178, demonstrate substantially lower levels of impurities. While still not being spectroscopic grade ethanol, these samples could well represent ordinary pharmaceutical grade ethanol. The authors claim these samples are “different”, however the evidence presented for this is weak to nonexistent.

Figure 1 presents one trace each for Nat mur and Nux vomica, each at 6C, 12C and 30C potencies. The traces are said to be “representative”, however with no information on repeatability or how the “representative” traces were selected, it is impossible to say whether there is any real difference between any of the six spectra.

Figure 2 purports to address this point, but then fails to present the necessary data. The legend declares that 10 samples of each of the six remedy preparations were analysed. The accepted way to present such data would be as mean absorbance ± standard deviation for each wavelength point, or at least for a representative selection of wavelength points. Statistical analysis could then be used to demonstrate whether or not there was a real difference between any of the remedies or potencies. However, the authors have instead chosen to present only two traces for each preparation, as “envelopes of differences”. The derivation of these traces is not explained, although we surmise that “extreme” high and low traces for each preparation were chosen to provide an impression of the range of results obtained. This is not an appropriate method of handling data of this nature, as most of the information is lost and statistical analysis is rendered impossible.

A further difficulty with figure 2 is that the upper (open circles) trace in the top graph of fig 2a (30C Nat mur) appears to be a duplicate of the upper (filled circles) trace in the top graph of fig 2b (30C Nux vom). Comparison with other traces of the two remedies indicates that this trace is really one of Nux vom, which has been duplicated into the Nat mur graph in error.

Paucity of data, ambiguity of presentation and lack of statistical analysis prevent any conclusions being drawn from the information in figure 2.
Comparison of figure 2 with figure 1 reveals that all six traces presented in figure 1 are taken from figure 2, in each case the filled-circles traces. If indeed the traces in figure 2 represent the extreme range of results obtained, this is startling, as the traces in figure 1 are stated to be “representative”. In addition, while it does appear that the Nux vom samples tended to demonstrate higher absorbances than the Nat mur samples (excluding the obvious mistake noted above), in two out of the three potencies the higher Nux vom trace from fig 2 has been chosen for inclusion in fig 1, thus exaggerating the apparent difference.

Figure 3 (b and c) again repeats the same six traces as figure 1, this time grouped by remedy. Presented in this way, it is clear that there is absolutely no difference between the three potencies of Nat mur, and that while variation between the Nux vom potencies is a little more pronounced, again all three appear to come from the same population. The same is true of the three potencies of “succussed ethanol” presented in fig 3a.

On simple visual inspection it does appear that there may be genuine differences between the three remedies (although no statistics are presented to allow this to be tested), with the Nat mur showing the lowest absorbtion and the Nux vom the highest, with the succussed ethanol lying somewhere between. Nevertheless, these differences are entirely consistent with small differences in purity of the ethanol stock used for preparation of the three remedies – small, that is, relative to the very high level of impurity evident in the “plain ethanol” sample presented alongside. This degree of variation in UV absorbance is entirely to be expected between different batches of pharmaceutical grade ethanol, which is not prepared with spectroscopic analysis in mind. The authors make no mention of having stipulated to Hahnemann Laboratories that all material sent to them should be prepared from the same stock bottle, and the data presented indicate that the different remedies, possibly prepared at different times, simply came from different bottles of ethanol.

We hope you will agree that these are very serious points, and it is regrettable they were not identified by your own scrutineering process. It is clear that the data presented are wholly inadequate to support the authors’ assertion that UV spectroscopy can differentiate between the two remedies, and between different potencies of the remedies. If the authors wish to test their assertion so that it can be substantiated it will be necessary to repeat the work from the beginning, ensuring that all samples used in the study are sourced from the same bottle of stock solvent, that all duplicate preparations for precision assessment are separately prepared de novo from the mother tinctures, and that sufficient data are generated to allow robust and valid statistical analysis of the results.

Yours faithfully,

Rolfe
JJM
Wilsontown
Pipirr

References:
1. Rao, M. L., Roy, R., Bell, I. R. & Hoover, R. (2007) The defining role of structure (including epitaxy) in the plausibility of homeopathy. Homeopathy 96, 175-182.
2. Sigma Aldrich catalogue, ACS spectrophotometric grade ethanol 95.0%, at www.sigmaaldrich.com/catalog/search/ProductDetail/SIAL/493511
3. Sigma Aldrich catalogue, USP/NF grade ethanol 190 proof, at www.sigmaaldrich.com/catalog/search/ProductDetail/ALDRICH/493538

That’s really interesting stuff. I’d picked up on some of your observations, but didn’t know enough about the technique to be sure I was wasn’t just making an arse of myself.

If you’re interested there’s a big discussion on this paper at the James Randi Foundation forum, where they’re dying for some input from knowledgeable Raman spec people:

forums.randi.org/showthread.php?t=88831

hawk-handsaw.blogspot.com/2007/08/very-bad-science.html

Hopefully you can see that the graphs are the same when presented side by side. Of course, there should be no need to go through such a rigmarole to compare graphs in the first place…

Much Corel Drawing this morning. Your eyes are pretty good, I would say. In each case in figure 1, the curves come from figure 2: so if envelope of difference means anything at all, then they’re comparing the most extreme examples, as you said.

The really interesting one, though, is where the ‘envelope of difference’ for 30c Nat Mur is the same as the comparison graph for 30c Nat Mur vs. Nux Vom, as described above.

Ben: do you have Corel Draw? If not, I can mail you a jpeg or some such.

Anyway, I think we can reasonably conclude that the UV-vis spectra in this paper show nothing whatsoever.

Just Googled for fun – the only place “envelope of difference” comes up is on the Randi forums where they are talking about this very paper – obviously not a real statistical term.

Anyway I’m glad its not just my eyes on the spectra in Fig 1 and 2 – I couldn’t really be sure as my bloody computer screen is too small!

It’s stranger than you think. I pratted about stretching and re-sizing images in Corel Draw, so I could directly compare curves. If you look at the graph in figure 2a, supposedly the envelope of difference for 30C Nat Mur, and put it on top of the curve in figure 1 that shows the difference between 30C Nat Mur and Nux Vom…they’re the same fucking graph! Absolutely identical! So the same graph is purporting to be two different things!

I haven’t got time to get into this any further right now, but it doesn’t bode well, does it?

There’s been a fair bit posted about this paper here:

forums.randi.org/showthread.php?t=88831

Seems there are some issues with the ethanol used as the ‘blank’.

Another thing about that ‘envelope of differences’ in Fig 2. If one curve showed the highest values and one showed the lowest values, then the curves wouldn’t cross. The curves do cross for the Nat Mur examples in Figure 2, which suggests that the ‘envelope of difference’ is something else. Perhaps they picked one spectrum that was quite high, and one that was quite low, and used those? This isn’t helped by the fact that there’s no key to explain what the open versus closed symbols on the curves mean.

I have been having another look at the paper and I’m partly wrong – it appears after all that they did test more than one prep of each – pity they forgot to put that in the methods section! Figure two does indeed show an “envelope of differences” for 10 preps of each homeopathic medecine – I’m not quite sure what an “envelope of differences” means but I’m guessing it is the highest and lowest reading at each wavelength amongst the 10 preps tested. And it appears that between 200 and 400 nm there is indeed quite a bit of scatter from prep to prep.

How does this apply when you compare the medicines to the succussed solvent only? – well I’ve no idea as they don’t actually tell you what the data used on those spectra (Figure 3) are – is each a “representative” spectra? is it an average of the 10 preps from Fig 2? Where are the stats?

Also if we then go back to Figure 1 which is supposed to show the differences between the two medicines I think I see something very dodgy. Look at the 30C comparison and then pull up Figure 2 alongside it – it could just be my eyes, but in Fig 1 30C it looks like the Nux vom line is an exact match shapewise with the upper line of the “envelope of differences” from Fig 2 and the Nat mur line is an exact match shapewise with the lower line from that “envelope of differences” in Fig 2 i.e. it appears they have picked “representative” spectra which are the extremes up and down from each series of 10 preps. Hardly surprising they are different if that is the case!