285 replies to this topic

[kmoody]

All vendor vessels were insufficient to stop by-product formation.​

 

​​I understood goodandcheap passed your quality control Kelsey, have you tested subsequently and found them to fail?

 

Yes, this is correct. No vendor has consistently met the product specifications they put on the label.

[Daniel Cooper]

 

Let's look at the bright side:

If it is really epoxides forming in c60oo, and if those were the reason for cancer in the study, we won't need lengthy animal experiments to address this issue. All we would need is:

1. proof that epoxides are forming in amounts that can cause cancer in those test animals (or if its not epoxides find what else is being formed)

2. reproduce the formation of epoxides in c60oo

3. add antioxidants to fresh c60oo using the same protocol used in 3. and see if epoxides form again, if no, yay, storage solution found (we now just need to find out how long it can be stored), if no, find another way to stop epoxide formation...

 

And only then, start again with animal experiments.

 

Agreed, but easier said than done. When we tried to force generate epoxides by light exposure the resulting formulation was acutely lethal in the test animals. So the question becomes whether or not it is worth optimizing the "perfect" dose that causes/exacerbates cancer. We are preferring a different course of study, trying to identify alternative formulations that preserve the efficacy but are resistant to epoxide formation. At some point we probably will need to do a dose response to determine how much contaminant is necessary to actually be toxic. Its just a messy (and expensive) series of studies to run.

 

 

 

Disturbing.  

 

If C60oo can form acutely lethal epoxides then even if it's formulated correctly and there are no epoxides at manufacturing then you have to get concerned about what conditions these compounds are exposed to in shipping and storage.  

 

 

Did you measure the level of epoxide formation after your light exposure?

 

 

[kmoody]

Disturbing.  

 

If C60oo can form acutely lethal epoxides then even if it's formulated correctly and there are no epoxides at manufacturing then you have to get concerned about what conditions these compounds are exposed to in shipping and storage.  

 

Did you measure the level of epoxide formation after your light exposure?

 

Precisely our concern. When we asked vendors about stability data (recall that 3 years is the listed shelf-life for most of these) there was no empirical evidence backing this number. Their claim was the powder is stable for 3 years (there was data for that). Their claim was the olive oil is stable for 3 years per manufacturer they sourced from (I have not seen this data). So their claim is the combination of powder and olive oil is also stable for 3 years.

 

Obviously this is completely wrong. The stability of the product can be (and probably is) different than the stability of the individual raw materials. So no real stability data exists (that I am aware of).

 

We have not been able to characterize or properly measure epoxide formation as I do not yet have a MS in my lab. I am hoping to get one soon that would allow us to answer some of these questions.

[Turnbuckle]

The animals died of within 1-3 days of toxicity from excessively light exposed C60-EVOO, an outcome we did not expect. 

 

 

That is rather shocking. So that gives an LD50 dose of what? A few ml/kg for irradiated solution?

 

BTW, I'd advise anyone making their own to store it in metal jars, as amber glass is not sufficient.

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[Daniel Cooper]

 

The animals died of within 1-3 days of toxicity from excessively light exposed C60-EVOO, an outcome we did not expect. 

 

 

That is rather shocking. So that gives an LD50 dose of what? A few ml/kg for irradiated solution?

 

BTW, I'd advise anyone making their own to store it in metal jars, as amber glass is not sufficient.

 

 

 

Sounds like we don't know LD50 since they did not measure the level of epoxides after their light exposure.

 

It might be helpful to know what dose wrt light exposure.  Room lighting?  High intensity discharge?  What levels, how long, source, .... those sorts of things.

[Daniel Cooper]

BTW - is using metal jars even safe?  Can we imagine the exposed metal surface in contact with the C60oo catalyzing epoxide formation?  I don't know but I'd probably think about that.

 

 

 

[lost69]

so to sum up:

 

no commercial vendors 100% safe

 

home made with metal jar or glass jar wrapped with aluminium foil is safe (i have no idea how to make it without any light)

 

then how do we take it with no light exposure?some light is needed to see the ml in the syringe 

 

what about infrared stuff to see in the dark?

 

[Daniel Cooper]

so to sum up:

 

no commercial vendors 100% safe

 

home made with metal jar or glass jar wrapped with aluminium foil is safe (i have no idea how to make it without any light)

 

then how do we take it with no light exposure?some light is needed to see the ml in the syringe 

 

what about infrared stuff to see in the dark?

 

 

I'd sum it up as "what we know is significantly outweighed by what we don't know".

 

WRT storing it, if I were still doing C60oo, I'd do amber glass jars inside some sort of light tight outer enclosure.  Say glass bottle inside metal can, stored in a "cool dry place".

 

For me, there's just too many uncertainties.

 

As far as how to fill your syringe, I don't believe you'd have enough time exposure to form many epoxides in any sort of reasonably light room.  Dim the lights if you are more cautious.

Edited by Daniel Cooper, 14 February 2017 - 09:47 PM.

[lost69]

i'm getting too many pros to stop it, i prefer to find a very safe way of doing it/taking it with monthly blood tests just for extra safety

 

it would be useful to set a safe level of light and exposure time if there is any

 

infrared glasses (in case they work i only saw this stuff on 007 movies) would be a perfect solution to avoid light

[Wilberforce]

I'm interested why none of the suppliers have switched to MCT oils per Turnbuckle's experiments.


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[ambivalent]

It would be useful to know how this light exposure conducted in the lab translates to practical light exposure the oil is likely to experience (hours in an amber bottle exposed to the sun, say, or left under a desk lamp). It is hard to imagine that the c60oo oil used by had not decayed at all, yet there was no sign of tumours in the rats (afaik). There must surely be a tipping point, a level of light exposure up to which c60oo is not harmful, but beneficial, beyond it and obviously it becomes toxic. Another question would be how likely is it that we experience benefits from oil that is carcinogenic e.g. would the fact that my skin softens the day after taking dose be indicative that the oil has not decayed significantly through epoxide reactions? Turnbuckle's red light experiments seemed to indicate, from recollection, a loss of aerobic effects from excessive red light exposure (in MCT Oil).

[Turnbuckle]

Turnbuckle's red light experiments seemed to indicate, from recollection, a loss of aerobic effects from excessive red light exposure (in MCT Oil).

 

 

A certain amount of red light improved exercise performance when the right additives were dissolved in the oil. CoQ10, for instance. It was likely that C60 was reacting with it and taking it into the mitochondria. It was also likely that C60 was reacting with the the FAs of the MCT oil, as the overall effect was slightly unpleasant.

[ambivalent]

Yes, I remembered that wrong: in your trial with the 30 minute red light exposure to MCT there was a drop off in benefit, but still benefit despite a rancid smell in the oil and in fact the c60oo mix showed no noticeable effects, in your experience, to the same exposure in taste or odour, which, I'd have thought, either suggests a greater resilience with the olive oil mix or an increased masking effect.

 

Given Kelsey's explanation that c60-concentration can be a marker of levels of toxic by-products, wouldn't we have expected greater variance in the concentration levels of the c60oo samples measured given the ease in formation of toxic by-products and the likelihood of different storage conditions post distribution from those submitting samples? It would seem in that case, by the results, assuming the manufacturers weren't in fact producing products with different levels of concentration of c60, that all the formation of these toxic by-products was solely down to storage and preparation conditions by the vendor, little subsequently by the users.

 

Also it would be useful to know, as a comparison, the concentration levels, as such the toxic by-product levels, recorded in the heavily light exposed oil which had such catastrophic effects on the mice.

[Turnbuckle]

Yes, I remembered that wrong: in your trial with the 30 minute red light exposure to MCT there was a drop off in benefit, but still benefit despite a rancid smell in the oil and in fact the c60oo mix showed no noticeable effects, in your experience, to the same exposure in taste or odour, which, I'd have thought, either suggests a greater resilience with the olive oil mix or an increased masking effect.

 

 

 

 

I doubt that olive oil has any greater resistance to degradation. FAs of olive oil have greater molecular weight and are thus less volatile, and one FA of MCT oil (capric acid) has a strong natural odor of rancidity. Thus breakdown of MCT oil will be much more apparent to the nose.

[ambivalent]

For perspective it is worth recalling Agevivo's home lifespan study:

 

On the bleak side all three mice were reported to have tumours at death despite (from Turnbuckle) a tumour rate expectation of one in four. Although it is not clear the mice died from the tumours and, possibly, given the age of a couple of them the expectation might have been more likely than 1 in 4. But it certainly seems quite likely or at least very possible, given subsequent studies, that c60oo played a role in tumour growth in the mice. 

 

However, the oil used was old, left over from the original Baati study*. Niner mentioned a sample was run on HLPC by Prof. Moussa's lab and suggested 'but I'm not sure that they could tell if the oil was rancid if they were just looking at a c60 signal'. In addition, from Kelsey's Moody's observations, there seems to be some ambiguity in the viability metric of the signal of c60 concentration levels given the presence of toxic bi-products - although presumably, if it met c60 concentration specs then this would indicate an absence of bi-products. On top of that there is still the storage of the oil when handled by Agevivo which, presumably, unless free from light and frozen would have degraded.

 

 

The mice were bought from a store at a few weeks old and believed to be 18 months of age (3 females) at the start of the study. From recollection the dosing was weekly, after an additional loading period, but may have increased at some point.

 

So the administration of the oil used in his experiment, which admittedly we don't know was significantly degraded, but would certainly, in light of current understanding, not be fit for purpose for running a similar experiment, saw the three 18 month old mice** live an additional 4, 13 and 13 months to respective ages of approximately 22 months and 31.5 months (x2)

 

 

 Anecdotally, the two oldest mice appear to have been quite exceptional in age.

 

 

 

*Does anyone know the storage conditions of the oil: during Baati/post Baati/Agevivo and indeed when it was made?

 

** possibly a month younger

Edited by ambivalent, 16 February 2017 - 12:25 PM.

[Turnbuckle]

 

 

 

 

*Does anyone know the storage conditions of the oil: during Baati/post Baati/Agevivo and indeed when it was made?

 

** possibly a month younger

 

 

You can see from the pictures in this post that the Baati sample had lost its red color and turned muddy by the time AgeVivo got it. That it was stored in a clear container suggests that it got a large dose of light over the years.

[ambivalent]

 

 

 

 

 

*Does anyone know the storage conditions of the oil: during Baati/post Baati/Agevivo and indeed when it was made?

 

** possibly a month younger

 

 

You can see from the pictures in this post that the Baati sample had lost its red color and turned muddy by the time AgeVivo got it. That it was stored in a clear container suggests that it got a large dose of light over the years.

 

 

Thanks Turnbuckle, that's certainly suggests the likelihood the mice were given degraded oil.

 

I tend not to bite but to the ill-informed remarker - try to be more useful and respond - it just comes across that you simply don't like what is being written but don't have a cogent or coherent response.

 

There isn't anything ill-informed there, the post is by and large a summary of the facts ad commentary of Agevivo's study in which the mice were very likely administered with degraded oil and lived for a substantial time while supplemented with it. If you contest the facts as written in the log to be wrong, then there is indeed a case for your ill-informed claim and you'd be wise to correct it but a splash of red paint serves no purpose, save self-indulgence.

Edited by ambivalent, 16 February 2017 - 07:36 PM.

[Huckfinn]

I've been making C60OO by simply shaking a dark coloured bottle wrapped either in aluminium foil or brown paper for a couple of weeks since about the last 1.5 years.

I use 99.95% pure from SES research.

I take one tbs/day most days.

And keep the 750ml bottle in a cupboard.

 

Is it an ok way to prepare it or should I switch to a magnetic stirrer?

I still have one unopened  little plastic "vial" of C60 from a year ago in the cupboard...would it still be ok or should I reorder it?

 

...Over this time the main difference I noticed is the remarkable fact that a beginning of white hair on the sides of my head stopped and has reverted to the original chestnut colour...

 

2 little points about EVOO:

-Chlorophyll should be an agent "preserving" the qualities of the OO (antioxydant) but only when the OO is kept in the dark. It's part of the green component of the oil. Better stainless steel containers than aluminium as they do it in the mediterranean area.

-Freezing OO might be detrimental as the vitamins in it might suffer, in particular Vit E.

Edited by Huckfinn, 17 February 2017 - 03:19 PM.

[Turnbuckle]

 

-Freezing OO might be detrimental as the vitamins in it might suffer, in particular Vit E.

 

 

Evidence?

[Huckfinn]

I cannot find you the source of the information, Turnbuckle.

 

A quick search in pubmed gave me (and it's not where I got it from I'm afraid):

"At the Purdue University Animal Disease Diagnostic Laboratory, it was recognized that hemolyzed porcine serum samples and frozen whole blood samples that were subsequently thawed for analysis were either deficient in or had no detectable amounts of vitamin E..... In humans, studies have evaluated the stability of vitamin E during blood collection and processing and during longterm storage of serum. These studies have concluded that vitamin E is relatively stable during most collection, processing, and storage conditions (up to many years when frozen at 270 C), but they do not directly address the stability of vitamin E when hemolysis has occurred......"

[zen]

 

No. The animals died of within 1-3 days of toxicity from excessively light exposed C60-EVOO, an outcome we did not expect. Because we did not expect it, we were gearing all our study endpoints to look at tumor burden and were unfortunately unable to do anything useful in terms of determining cause of death.

​1-3 days from consumption to death is scary.
What do you mean by "excessively light exposed"? Could you provide more details please?
​Have you tried to repeat this lethal experiment with the home made C60 Olive Oil?
 

[ambivalent]

What do you mean by "excessively light exposed"? Could you provide more details please?
​

 

I agree this would be good to know.

 

 

It seems to me from a rather simplistic standpoint that c60oo compensates for these toxins, to a point - enabling the repair of the damage they create. I think the Baati study and Agevivo's home-study lend some evidence to this (given in Baati's case the state of the oil handed to Agevivo).

 

That I know of we have five c60oo data points of mice administration:

 

Original Baati oil: made under ideal conditions, possible degradation over the course of administration. Exceptional outcome longevity, cancer. 

 

ichor 1: Prepared as above, improbable degradation. Exceptional results in slowing down tumour growth in leukaemia mice.

 

Ichor 2: 3rd party (SES): A degraded sample outlier in either storage (the company advocates a long shelf life) or possibly in preparation (dissolving via ultra sound). Failed Ichor's QC although it is quantitatively unestablished to what extent that sample failed compared to other 3rd party samples. Outcome: accelerated tumour growth in leukaemia mice, shorter lifespan than the leukaemia control mice.

 

Ichor 3: A clear sample outlier with the oil exposed to excessive light. Substantial epoxide formations. Unknown how excessive the levels of epoxides formed were or how the exposure scales to typical environmental risk factors in warehouse, transport and user storage conditions likely to be experienced by the distribution of c60oo users. Outcome: acute mortality in mice: 1-3 day lifespan.

 

Agevivo: Prepared under ideal conditions but years earlier, light exposed for an unknown period and appears visually to be heavily degraded. Again an expected outlier in the samples of oil consumed by c60oo users. Outcome: probable life extending and tumour creating effect on 17-18 month old mice.   

[kmoody]

​1-3 days from consumption to death is scary.

What do you mean by "excessively light exposed"? Could you provide more details please?
​Have you tried to repeat this lethal experiment with the home made C60 Olive Oil?

 

The sample was artificially exposed to high levels of UV light from a power source. The source material we used for this was administered as was the UV light exposed in two different arms of the study. The UV exposed was toxic and resulted in deaths 1-3 days from consumption.

 

This suggests that there is a level of toxic by-product formation that is lethal. More importantly, this is suggestive that even lower levels of by-product formation could be sub-lethally (and perhaps even sub-clinically) toxic.

[Daniel Cooper]

 

​1-3 days from consumption to death is scary.

What do you mean by "excessively light exposed"? Could you provide more details please?
​Have you tried to repeat this lethal experiment with the home made C60 Olive Oil?

 

The sample was artificially exposed to high levels of UV light from a power source. The source material we used for this was administered as was the UV light exposed in two different arms of the study. The UV exposed was toxic and resulted in deaths 1-3 days from consumption.

 

This suggests that there is a level of toxic by-product formation that is lethal. More importantly, this is suggestive that even lower levels of by-product formation could be sub-lethally (and perhaps even sub-clinically) toxic.

 

 

 

Do we know the wavelength and lux level of illumination incident on the C60oo sample?

 

"UV" and "High Intensity" are rather loose terms.

[Graviton]

 

​1-3 days from consumption to death is scary.

What do you mean by "excessively light exposed"? Could you provide more details please?
​Have you tried to repeat this lethal experiment with the home made C60 Olive Oil?

 

The sample was artificially exposed to high levels of UV light from a power source. The source material we used for this was administered as was the UV light exposed in two different arms of the study. The UV exposed was toxic and resulted in deaths 1-3 days from consumption.

 

This suggests that there is a level of toxic by-product formation that is lethal. More importantly, this is suggestive that even lower levels of by-product formation could be sub-lethally (and perhaps even sub-clinically) toxic.

 

What was the age of rats that UV radiated C60 were administrated? It reminds that Agevivo's rats were in about mid age while Baati's rats were early stage of rats' lifespan. Isn't there any peculiarity of their response on C60oo? Niner previously discussed that the period of lifespan given with C600 can be a factor for C60 effects. Tumor burden might be different between two age groups.

[Turnbuckle]

 

 

 

​

 

The sample was artificially exposed to high levels of UV light from a power source. The source material we used for this was administered as was the UV light exposed in two different arms of the study. The UV exposed was toxic and resulted in deaths 1-3 days from consumption.

 

This suggests that there is a level of toxic by-product formation that is lethal. More importantly, this is suggestive that even lower levels of by-product formation could be sub-lethally (and perhaps even sub-clinically) toxic.

 

What was the age of rats that UV radiated C60 were administrated? It reminds that Agevivo's rats were in about mid age while Baati's rats were early stage of rats' lifespan. Isn't there any peculiarity of their response on C60oo? Niner previously discussed that the period of lifespan given with C600 can be a factor for C60 effects. Tumor burden might be different between two age groups.

 

 

 

As kmoody said above, these deaths weren't from cancer. These were due to an acute toxic response. Too fast for cancer to play a part. This has been known for some time. One of the authors of the original rat study-- Fathi Moussa--pointed out ten years ago that C60 solutions become toxic when exposed to UV or even visible light--

 

Therefore, if pristine C60 is absolutely nontoxic under dark conditions, this is not the case under UV-Visible irradiation and in the presence of O2 where fullerene solutions can be highly toxic through 1O2 formation. 

 

https://www.ncbi.nlm...pubmed/18217343

 

 

Some have taken his words "C60 is absolutely nontoxic" out of context. But Moussa points out several examples of toxicity. From the paper I linked to above--

 

They show that in contrast with pristine C60, some derivatives can be highly toxic (Fig. 1).

Furthermore, their effects in vitro generally do not reflect exactly their in vivo behaviour. The

first study on the toxicity of C60 derivatives was conducted by Yamago et al.19 After a single

intra-peritoneal injection of 0.5 g/ kg of bodyweight of a water-soluble methanofullerene (Fig.

1C), the mice survived for one week. One year later Rajagopalan et al. studied the pharmacokinetics

of another water-soluble methanofullerene (Fig. 1D).49 Although the in vitro toxicity

of this derivative was quite low,50 it appeared highly toxic in vivo.49 At a dose of 15 mg/kg, this

derivative seemed well tolerated. However, a dose of 25 mg/kg resulted in shortness of breath

and violent movement of the rats, followed by death within 5 min of dosing.49 We also observed

in our laboratory a similar behaviour in mice treated with a cationic C60-derivative for a

dose as small as 2 mg/kg of bodyweight (unpublished work). 

 

 

 

 

Edited by Turnbuckle, 18 February 2017 - 12:09 PM.

[zen]

 

​1-3 days from consumption to death is scary.

What do you mean by "excessively light exposed"? Could you provide more details please?
​Have you tried to repeat this lethal experiment with the home made C60 Olive Oil?

 

The sample was artificially exposed to high levels of UV light from a power source. The source material we used for this was administered as was the UV light exposed in two different arms of the study. The UV exposed was toxic and resulted in deaths 1-3 days from consumption.

 

This suggests that there is a level of toxic by-product formation that is lethal. More importantly, this is suggestive that even lower levels of by-product formation could be sub-lethally (and perhaps even sub-clinically) toxic.

 

How long was the sample exposed to the UV light?
​Was the sample in the bottle?
​If in the bottle, was the bottle full or not?
​In other words, was the sample not only exposed to the UV light but also to the large amount of oxygen?

If not only UV light but also an oxygen is needed to achieve toxicity then, maybe, storing the C60-OO in small amber glass bottles which are completely filled up, in the refrigerator (obviously there is not much light there) is enough to make the C60-OO safe?
 

[Graviton]

 

All vendor vessels were insufficient to stop by-product formation.​

 

​​I understood goodandcheap passed your quality control Kelsey, have you tested subsequently and found them to fail?

 

Yes, this is correct. No vendor has consistently met the product specifications they put on the label.

 

I am confused with Kmoody's study.

What do you mean by "All vendor vessels were insufficient to stop by-product formation". It is not clear what is insufficient to stop by-product formation. Do you mean by "C60 amount" by HPLC?

Which below number(s) describe the above statement?

1. All vendor C60 olive oil has not sufficient amount of C60 as their label state? Wasn't there analysis of quality control of experimental C60 amount comparing to the amount stated in different vendors' labels in the previous thread? 

2. Did you test all vendors' C60 epoxide levels?

3. Did any vendor's C60 olive oil cause the growth of tumor or toxicity other than SES olive oil?

[Empiricus]

I posted here about how mega-dosing NR reduced back ache and nausea (symptoms I strongly suspect were kidney-injury related).  

 

What could those of us concerned about our past exposure to likely toxic c60 brews do by way of tumour prevention?  What supplements should we consider taking?  Perhaps this topic merits its own thread.

Edited by Empiricus, 19 February 2017 - 08:41 AM.

[kmoody]

What do you mean by "All vendor vessels were insufficient to stop by-product formation". It is not clear what is insufficient to stop by-product formation. Do you mean by "C60 amount" by HPLC?

Which below number(s) describe the above statement?

1. All vendor C60 olive oil has not sufficient amount of C60 as their label state? Wasn't there analysis of quality control of experimental C60 amount comparing to the amount stated in different vendors' labels in the previous thread? 

2. Did you test all vendors' C60 epoxide levels?

3. Did any vendor's C60 olive oil cause the growth of tumor or toxicity other than SES olive oil?

 

We took empty vessels from each of the vendors and tested their ability to prevent light mediated degradation of C60oo. None of them were effective at blocking out the light at appreciable levels.

 

I would have to review my notes for the exact details, but I believe we put in fresh C60oo in each vessel then exposed to light. In one grouping it was UV exposed, in another grouping it was left out for some period of time in ambient light.

 

1. This is correct. No vendor has consistently met their label claims.

2. We cannot directly measure epoxide formation yet. We have almost finished developing an assay in house that will allow us to quantify the presence of epoxides. That work is ongoing and should be done soon.

3. We have not tested other vendor C60oo in vivo. It did not make sense to me to poison animals for no scientific reason.

 

Broadly, our interest is not characterizing the products sold by online vendors. We know the QC is of poor quality or does not exist at all, and we know that light exposure causes the formation of what we believe are toxic epoxides. What we are more interested in is finding ways to prevent epoxide formation, or to develop a formulation that preserves the efficacy of dark C60oo but is resistant to degradation from light. For this reason, we have done some, but not extensive, characterization of the various products of 3rd party vendors.

 

Note that these are laborious and expensive projects to conduct, particularly when we get animals involved. I also have reservations about poisoning my mice to prove vendors are selling toxic products. We have shown they do not meet the basic claims listed on the label. It is their burden to prove out their product.

Edited by kmoody, 19 February 2017 - 07:39 PM.