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C60 in olive oil mediated life extension: Scientific discussions

c60 buckyballs lifespan baati moussa fullerenes

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#31 geo

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Posted 21 April 2012 - 07:12 PM

First post here but I read the forum for a long time. While searching the web for information concerning Fullerene, I found that something is suspicious with the paper. Check it out.
http://pipeline.cora...s_a_problem.php
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#32 Googoltarian

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Posted 21 April 2012 - 07:34 PM

Its interesting how C60 treated rats all die in very small time frame, when compared to other groups. Hayflick limit?
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#33 Elus

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Posted 21 April 2012 - 08:13 PM

DMT: Accept your mortality.


Don't tell me what to do - I will not accept it. I will rebel against it. More bluntly put: I will seek immortality.

It's your complacency with death that's holding us back. It's the pro-aging trance.

On topic: Does anyone find it strange that the oil alone resulted in lifespan extension?

Edited by Elus, 21 April 2012 - 08:15 PM.

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#34 Krell

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Posted 21 April 2012 - 08:57 PM

Some more details on the Rat Shack Forum

http://www.ratshackf...daa2ec7&start=0

#35 hav

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Posted 23 April 2012 - 02:43 PM

I'm wondering if the fullerene might not be the active longevity ingredient. Maybe it’s just the transport. And dissolving it in olive oil also allows some olive oil molecules to penetrate the fullerene sphere and get transported to places by the fullerene that orally ingested olive oil wouldn't make it to. Like deep into many cells throughout the body even those across the bbb. It that's right, all the longevity effects observed are just an amplification of the effects of the olive oil. And maybe other oils might be even better. Like fish oil for instance. And maybe the transport might work for other longevity molecules that might also be able to be dissolved into the oil. Wonder if cycloastragenol or epithalon might work. Of course this is all just pure speculation on my part. All the studies say the mechanism of operation is unknown. But the possibilities are exciting.

Howard

#36 Hebbeh

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Posted 23 April 2012 - 04:08 PM

Interesting theory. If so, the possibilities are endless. Exciting possibilities.

#37 Krell

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Posted 23 April 2012 - 06:36 PM

from the
FIGHT AGING! NEWSLETTER
April 23rd 2012

A PUZZLING FULLERENE STUDY Tuesday, April 17, 2012 http://www.fightagin...erene-study.php A research group is claiming that fullerenes (C60, ingested and injected) greatly extend life span in rats; this is meeting with some considerable skepticism, given the degree of life extension and the lack of a plausible mechanism. "In the current study researchers fed the molecule dissolved in olive oil to rats and compared outcomes to a control group of rats who got plain olive oil. The main question they wanted to answer was whether chronic C60 administration had any toxicity, what they discovered actually surprised them. ... Here we show that oral administration of C60 dissolved in olive oil (0.8 mg/ml) at reiterated doses (1.7 mg/kg of body weight) to rats not only does not entail chronic toxicity, but it almost doubles their lifespan. ... The estimated median lifespan (EML) for the C60-treated rats was 42 months while the EMLs for control rats and olive oil-treated rats were 22 and 26 months, respectively. Using a toxicity model the researchers demonstrated that the effect on lifespan seems to be mediated by 'attenuation of age-associated increases in oxidative stress'." So what might be going on here? The average life span of the Wistar rats used is 2-3 years (24 - 36 months). This was a small study size, but that's not so important in determining whether you have an actual means of life extension if you can show that any of your study group lived much longer than usual - but it is important when it comes to the degree of life extension. If the study group is small, as it is here, using only a handful of rats, then the size of the effect can be much more readily distorted by chance. This line in the paper jumped out at me: "Before C60 administration, the rats were fasted overnight but with access to water." If they failed to fast the control group, then we're looking at yet another study that failed to control for calorie restriction, and this is actually largely an intermittent fasting study - which has certainly been shown to extend life in rats.


To subscribe or unsubscribe from the Fight Aging! Newsletter, please
visit http://www.fightaging.org/newsletter/

#38 Junk Master

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Posted 23 April 2012 - 08:13 PM

Intermittent fasting has never been shown to DOUBLE lifespan in rats. In fact, most intermittent fasting studies with rats show no more benefit than plain old CR.
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#39 DeadMeat

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Posted 24 April 2012 - 12:11 AM

I think 54 grams will last a whole year... for 1 big rat.


Something like that would be the case if you dosed said critter every day for a year(and without inter-species scaling). But they didn't dose every day all the time in the study.

The rats were housed three per cage and acclimated for 14 days, before dosing. Three groups of 6 rats (10 months old, weighing 465 +- 31 g) were administered daily for one week, then weekly until the end of the second month and then every two weeks until the end of the 7th month, by gavages with 1 ml of water or olive oil or C60 dissolved in olive oil (0.8 mg/ml), respectively. The rats were weighed before each dosing. Routine observations following official recommendations [27] were made on all animals inside and outside the cage once a day throughout the study for signs of departure from normal activity, morbidity and mortality.


For 77 kg of <:3)~~ one dose would be 1.7*77 = 131 mg. In the study they did 1 week every day, then 7 weeks weekly, then 20 weeks every two weeks. Then they stopped dosing and waited for the rats to die naturally which gives figure 3a. So 24 doses. 24*0.131 = 3.14 gram. Although I have no idea what would be optimal of course.

#40 PWAIN

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Posted 24 April 2012 - 01:19 AM

For 77 kg of <:3)~~ one dose would be 1.7*77 = 131 mg. In the study they did 1 week every day, then 7 weeks weekly, then 20 weeks every two weeks. Then they stopped dosing and waited for the rats to die naturally which gives figure 3a. So 24 doses. 24*0.131 = 3.14 gram. Although I have no idea what would be optimal of course.


If a rat lives for 3 years and a human for say 75 years, then a day for a rat may be equivalent to 25 days for a human? But then does a rat sleep 25 times in a 24 hour period? Really frequency and dose amount in humans is probably more guess work at this stage. How long to continue is also in question since from a rats perspective, they were taking this stuff for about a quarter of their normal life span.

Anyone else curious about the olive oil alone giving a lifespan boost too? Surely there are other lifespan studies with olive oil? Why would this one be different, is it different?

I am very hopeful but also quite suspicious.

#41 s123

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Posted 24 April 2012 - 06:52 PM

The paper has two main problems.

1. Between figure 3a and the reported results is a discrepancy of 8, 16, and 20 months for control, olive oil, and olive oil + C60 treated mice, respectively.

2. The histological picture for GAop and GAip (see figure 4) are identical (fraud or sloppy mistake?)
http://pipeline.cora...s_a_problem.php
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#42 Mind

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Posted 24 April 2012 - 08:22 PM

Sven, could you explain point 1 in a little more detail or is there somewhere we can look at the figures. A 20 month discrepancy would essentially nullify the 42 month life extension result.

#43 zorba990

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Posted 25 April 2012 - 04:16 AM

Interesting stuff. Can't wait for the clarification on mechanism of action.
Any thoughts on this? Looks a bit spammy but I'm real curois to know if the 2009 date is real or just an attempt to back date this in light of new findings...
http://smarteconomy....-to-market.html

#44 s123

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Posted 25 April 2012 - 11:52 AM

Sven, could you explain point 1 in a little more detail or is there somewhere we can look at the figures. A 20 month discrepancy would essentially nullify the 42 month life extension result.


"The estimated median lifespan (EML) for the C60-treated rats was 42 months while the EMLs for control rats and olive oil-treated rats were 22 and 26 months, respectively."

Then you look at figure 3a and you see that according to this figure all C60-treated rats are still alive! They only start dying after 60 months and mean lifespan is about 62 months. Thus the mean lifespan is 20 months longer in the figure than what is mentioned in the text. Same problem for the other groups.

Fig 3a Text
C60 62 42
Olive oil 42 26
Water 30 22

One can add two other problems in this paper:

1. I've failed to find any study showing a Wistar rat living to anywhere near 62 months.
2. The lifespan curve for C60 rats is suspiciously rectangular (James Fries would have loved it).

Below I have given examples of typical lifespans of Wistar rats from different studies. The data reported in the text fall within acceptable limits but those from figure 3a are incredibly higher than anything reported before.


From Walford and Weindruch, 1988 (Wistar rats, the strain used in the C60 experiment). R = Restricted and C = Control Lifespan in months.

R: 34(f), 31(m)
C: 31(f), 26(m)

R: 42
C: 33

R: 19
R: 39
R: 39
C: 33

R: 36
R: 38
C: 35

R: 41
C: 33

R: 39
C: 22

R: 35
C: 28

R: 46
C --> R: 41
R --> C: 42
C: 39

From van der Rijst et al. (1955). Voeding 16: 708.

Female: 22 months and Male: 20 months

From Shimokawa et al. (2003) FASEB J 17(9): 1108-1109.

C: 36 months
R: 44 months
Antisense GH transgene ©: 39 months
Antisense GH transgene ®: 47 months

This paper is either extremely sloppy or outright fraud.


#45 PWAIN

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Posted 25 April 2012 - 02:21 PM

So what is the Kaplan Meier estimator and how does it work? My understanding is that this is not just a straight median value. From the paper:


The survival distributions for C60-olive oil-treated rats and
controls were estimated by the non-parametric Kaplan-Meier
estimator (Fig. 3) and compared by a log-rank estimated test. The
estimated median lifespan (EML) for the C60-treated rats was 42
months while the EMLs for control rats and olive oil-treated rats
were 22 and 26 months, respectively. These are increases of 18 and
90% for the olive-oil and C60-treated rats, respectively, as compared
to controls

I also wonder if these are related to the lifespan since the commencement, midpoint or completion of treatments?

As for wistar rats not living to 62 months, isn't that the whole point, treatment with C60 is the supposed cause. I completely fail to see your point.

I was also under the impression that when pushing the boundaries of lifespan, things really square off - look at humans that reach 115 years of age - things really square off.

Sure I think we should be suspicious but I really don't see any actual evidence of fraud as of yet. A mistake with a slide image, a few misunderstandings do not a fraud make. We need this replicated urgently but other than that, I think we have something that looks promising. If this really pans out, the implications would rock the world.

Sven, could you explain point 1 in a little more detail or is there somewhere we can look at the figures. A 20 month discrepancy would essentially nullify the 42 month life extension result.


"The estimated median lifespan (EML) for the C60-treated rats was 42 months while the EMLs for control rats and olive oil-treated rats were 22 and 26 months, respectively."

Then you look at figure 3a and you see that according to this figure all C60-treated rats are still alive! They only start dying after 60 months and mean lifespan is about 62 months. Thus the mean lifespan is 20 months longer in the figure than what is mentioned in the text. Same problem for the other groups.

Fig 3a Text
C60 62 42
Olive oil 42 26
Water 30 22

One can add two other problems in this paper:

1. I've failed to find any study showing a Wistar rat living to anywhere near 62 months.
2. The lifespan curve for C60 rats is suspiciously rectangular (James Fries would have loved it).

Below I have given examples of typical lifespans of Wistar rats from different studies. The data reported in the text fall within acceptable limits but those from figure 3a are incredibly higher than anything reported before.


From Walford and Weindruch, 1988 (Wistar rats, the strain used in the C60 experiment). R = Restricted and C = Control Lifespan in months.

R: 34(f), 31(m)
C: 31(f), 26(m)

R: 42
C: 33


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#46 Turnbuckle

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Posted 25 April 2012 - 03:36 PM

So what is the Kaplan Meier estimator and how does it work? My understanding is that this is not just a straight median value. From the paper:


The survival distributions for C60-olive oil-treated rats and
controls were estimated by the non-parametric Kaplan-Meier
estimator (Fig. 3) and compared by a log-rank estimated test. The
estimated median lifespan (EML) for the C60-treated rats was 42
months while the EMLs for control rats and olive oil-treated rats
were 22 and 26 months, respectively. These are increases of 18 and
90% for the olive-oil and C60-treated rats, respectively, as compared
to controls

I also wonder if these are related to the lifespan since the commencement, midpoint or completion of treatments?


Since they started with ten month old rats and early deaths would thus not have been included, that must have something to do with it.

As for wistar rats not living to 62 months, isn't that the whole point, treatment with C60 is the supposed cause. I completely fail to see your point.


Exactly.

I was also under the impression that when pushing the boundaries of lifespan, things really square off - look at humans that reach 115 years of age - things really square off.


Except for two data points, the other curves would also be rather steep. I suspect this is more the product of too few data points than anything else. And if you were faking the data, you would go to extra lengths to eliminate any unusual aspect of the curves. On the other hand, a steeper curve is more likely to be associated with curing a disease than with an increase of lifespan. See fig. V2 at http://research.jax....vLifespan1.html

Edited by Turnbuckle, 25 April 2012 - 03:49 PM.


#47 geo

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Posted 25 April 2012 - 03:41 PM

Fraud or not, it is obvious that the experiment must be repeated. Thus, we have to wait a few more years. Let us hope that there will be scientists (with the necessary financial resources) willing to repeat the experiment.

#48 manic_racetam

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Posted 25 April 2012 - 06:08 PM

.......
"Initially, the hydrophobicity of C60 (water solubility <10-9mg/L) was thought to limit its interactions with biological systems and oral administration, skin application, or injection of C60 in rats and other cellsystems revealed no acute toxicity. However, once C60 is introduced into water (either via solvents or by extensive stirring) it forms stable nanoscale suspended aggregates known as fullerene water suspensions (FWS) or nC60 and becomes biologically active. nC60 is highly toxic to eukaryotic cell lines, Daphnia magna and fish."

In other words, any animal/human who is/were to ingest C60 better well make sure that it is suspended in oil like the authors of the life extending 2012 study did (study linked again for good measure: http://extremelongev...0-Fullerene.pdf) and abstain from drinking water close to the ingestion of the compound.. that being said humans are roughly 60-70% water.. it is a tricky but potentially extremely positive situation in relation to life extension.
....


So maybe an oil suspension with intramuscular injection would be a good option for increased safety over oral routes?

#49 Mind

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Posted 25 April 2012 - 08:29 PM

62 months is extraordinary-blow-the-roof-off-super-duper-fantastic lifespan extension for these rats. Extraordinary claims demand extraordinary proof. We should be very skeptical of such a small sample size and such a dramatic result. No one should assume much at this point. Definitely need further testing with larger sample sizes.
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#50 s123

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Posted 26 April 2012 - 12:46 AM

62 months is extraordinary-blow-the-roof-off-super-duper-fantastic lifespan extension for these rats. Extraordinary claims demand extraordinary proof. We should be very skeptical of such a small sample size and such a dramatic result. No one should assume much at this point. Definitely need further testing with larger sample sizes.



Indeed. According to figure 3a the lifespan was increased by 12 months by giving the rats olive oil alone! I'm sure nobody on this forum doubts that olive oil is healthy but a lifespan increase of 12 months is beyond credibility. However, if one looks at the text the lifespan difference is only 4 months. Which data are correct???

They started with 10 month old rats, probably bought ones. Now, I've been told that when you commercially order 10 month old animals and they don't have these in stock, they will instead give you what they have that is closest by your specified age. Of course, they tend to give younger and thus cheaper animals. This is another potential weakness in the study.
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#51 Turnbuckle

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Posted 26 April 2012 - 11:53 AM

A patent application by researcher Dr. Christopher Kepley suggests one mechanism by which fullerenes might extend life: Method for Inhibiting the Build-up of Arterial Plaque by Administering Fullerenes

http://appft.uspto.g...=DN/20110190251

And what good is long life if you must live it bald?

Fullerene nanomaterials potentiate hair growth. "As seen in Figure 2, A and B, mice injected with fullerene derivatives had accelerated hair growth as compared with those mice receiving vehicle only."

http://144.206.159.1...25/12825719.pdf

Dr. Kepley: http://www.lunainnov...2008_Kepley.htm

Edited by Turnbuckle, 26 April 2012 - 11:55 AM.


#52 geo

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Posted 26 April 2012 - 12:29 PM

Very interesting news. However, it seems that there is a question which is more crucial than the possible beneficial effects of Fullerene. The quesions is, what are the side effects of Fullerene. One can consume something that MAY be helpful, but I would not consume something that MAY be harmful. I read (somewhere) that if Fullerene is mixed with water, it is assumed to be toxic (it accumulates in the brain). Thus, a first thing that needs to be clarified, is the potential damage to the health of anyone that supplements Fullerene. From what I have read, I cannot reach an opinion.
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#53 AdamI

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Posted 26 April 2012 - 12:32 PM

There was a company that have manage to make a C60 that was not toxic in the water... soo guess you have to find out which one. I'm sure someone here knows it

#54 Turnbuckle

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Posted 26 April 2012 - 12:59 PM

Very interesting news. However, it seems that there is a question which is more crucial than the possible beneficial effects of Fullerene. The quesions is, what are the side effects of Fullerene. One can consume something that MAY be helpful, but I would not consume something that MAY be harmful. I read (somewhere) that if Fullerene is mixed with water, it is assumed to be toxic (it accumulates in the brain). Thus, a first thing that needs to be clarified, is the potential damage to the health of anyone that supplements Fullerene. From what I have read, I cannot reach an opinion.


Comparative pulmonary toxicity assessments of C60 water suspensions in rats: few differences in fullerene toxicity in vivo in contrast to in vitro profiles.

http://www.ncbi.nlm....pubmed/17630811

#55 geo

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Posted 26 April 2012 - 01:00 PM

A paper that covers the issue of Fullerene toxicity is the following:

http://toxsci.oxford.../2/162.full.pdf

The "conclusions" section is not very helpful for me (I am not an expert, but rather the opposite). All I understand is that more recearch is needed.

#56 Turnbuckle

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Posted 26 April 2012 - 01:12 PM

A paper that covers the issue of Fullerene toxicity is the following:

http://toxsci.oxford.../2/162.full.pdf

The "conclusions" section is not very helpful for me (I am not an expert, but rather the opposite). All I understand is that more recearch is needed.


The in vivo section makes it look rather safe, at least in the short term.

#57 PWAIN

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Posted 27 April 2012 - 01:50 AM

Very interesting news. However, it seems that there is a question which is more crucial than the possible beneficial effects of Fullerene. The quesions is, what are the side effects of Fullerene. One can consume something that MAY be helpful, but I would not consume something that MAY be harmful. I read (somewhere) that if Fullerene is mixed with water, it is assumed to be toxic (it accumulates in the brain). Thus, a first thing that needs to be clarified, is the potential damage to the health of anyone that supplements Fullerene. From what I have read, I cannot reach an opinion.


8bitmore's post here seems to give some indication of how this may be less harmful than water solution suspended C60:

http://www.longecity...857#entry511857

The original purpose of the testing was to see what damage was done to the mice. The expectation was build up of C60 in tissue, damage and early death. The researchers were surprised when quite the opposite happened. They found that there was no buildup of c60 in the tissues and that the c60 was flushed in a matter of hours. They found no tissue damage and they found considerable life extension.

I see only 2 possibilities here:

1. deliberate fraud
2. incredible discovery

Further testing will either rule out 1 or confirm it.

#58 PWAIN

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Posted 27 April 2012 - 02:07 AM

Indeed. According to figure 3a the lifespan was increased by 12 months by giving the rats olive oil alone! I'm sure nobody on this forum doubts that olive oil is healthy but a lifespan increase of 12 months is beyond credibility.



Yes, this is a concern and certainly worth investigating. I asked on another thread if anyone knows of any other lifespan study with olive oil but have not had any reply. It really seems unlikely and raises a red flag but until I see other studies of lifespan with olive oil, I'll reserve judgement.

However, if one looks at the text the lifespan difference is only 4 months. Which data are correct???



I believe I addressed this earlier in this thread.

They started with 10 month old rats, probably bought ones. Now, I've been told that when you commercially order 10 month old animals and they don't have these in stock, they will instead give you what they have that is closest by your specified age. Of course, they tend to give younger and thus cheaper animals. This is another potential weakness in the study.


This may be true but is highly speculative as you have no evidence either way. Even if we deduct the full 10 months off the measured lifespans, this would still be an incredible result so the impact is hardly changed. 2 to 4 months would be more likely. I don't this this would be the answer to this unexpected result - outright fraud would be more likely but has not been shown to be the case as of yet so we have to go with the results for now.

Edited by PWAIN, 27 April 2012 - 02:09 AM.


#59 niner

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Posted 27 April 2012 - 02:35 AM

I see only 2 possibilities here:

1. deliberate fraud
2. incredible discovery


3. gross incompetence.

Either way, we need replication. In the meanwhile, there are the other reports in the literature noting at least some degree of life extension from fullerenes. Something is happening here, we just don't know what. I kind of doubt that one little n=6 long term tox study from a no-name lab is enough to trigger a large, gold-standard experiment like the NIA's Interventions Testing Protocol. There would probably need to first be a reasonable-sized lifespan study with promising results. Thus it will probably be years before high quality evidence is available.

This result has stirred the waters too much for it to just fade away. Someone will do a bigger study. They've probably already ordered the C60. It took two weeks of stirring to get the C60 dissolved in the olive oil, so the adventurous among us should probably ask Santa for a good magnetic stirrer.

#60 Turnbuckle

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Posted 27 April 2012 - 11:25 AM

It took two weeks of stirring to get the C60 dissolved in the olive oil, so the adventurous among us should probably ask Santa for a good magnetic stirrer.


I tried C60 twenty years ago for an industrial application and I saw how difficult it was to dissolve in most solvents, so I was a bit surprised it went into vegetable oil so easily for these researchers. Anyway, I ordered another sample and yesterday added 400 mg to 500 ml of extra virgin olive oil, and within two or three hours, the light green of the oil had begun to darken. After the occasional hand shaking and 20 hours, about 95% dissolved (just eyeballing) and it's now a greenish whiskey color, about what you'd expect if you added the magenta color of a C60-toluene solution to the light green of the olive oil.

Edited by Turnbuckle, 27 April 2012 - 11:28 AM.






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