113 replies to this topic

[docmaas]

The only problem I see with this and the train of thought it is following is the assumption that we don't want the sulfation and glucouronidation to occur. It seems that there are at least some who are familiar with these issues that indicate that the glucouoronidated form may be in that form specifically for transport to other organs who will in turn convert it back. If that is the case then swamping the gut may actually be obstructing a useful pathway rather than aiding it. Note I'm not familiar with this whole process this is just a summary of what I think I have read here.

Mike

Yes I have been wondering about this. I have tried to find some articles but can't find anything. It is a very important question that needs to be answered. Do similar chemical structures to glucouoronidated resveratrol or sulfanated resveratrol get converted back to thier active form in cells?

Isn't sulfation and glucouronidation merely delayed by "swamping the gut"?

Yes, but it's that delay that results in higher plasma levels of native resveratrol, which is what you want. Eventually, it will all be conjugated and eliminated.

But the crux of the matter is whether this assumption is correct or not. If we avoid glucouronidation and sulphation we may also be shortstopping further bioprocesses that would otherwise use the conjugated products in as yet not understood ways. OTOH if they are bound to be conjugated regardless maybe the worst that happens is an unnecessary delay. Have tagged molecules been analyzed for their activity of either resv or its conjugates?

Mike

Edited by docmaas, 10 February 2008 - 12:02 AM.

[niner]

How about some piperine to help with those gut enzymes?

It wouldn't hurt, (unless you have something else in your system that you'd really like to glucuronidate) but I don't know how much it will help. Sulfation is a bigger factor in taking resveratrol out, but glucuruonidation is also pretty significant. I don't know if piperine affects the gut in the same way as the liver or not.

By what method is piperine increasing curcumin blood levels?

edit. I think piperine does work in the gut.

What about swamping the gut with other substances that are sulphated, like curcumin.

http://jn.nutrition....full/134/8/1948

Piperine inhibits glucuronidation, generally speaking. From the paper you cite, it looks like for EGCG, it's effective in the gut in mice. In that model, it inhibits glucuronidation in the gut 40%, but it doesn't inhibit glucuronidation of EGCG in vitro with liver microsomes, which is pretty suggestive that it's not working in vivo in the liver, for EGCG in mice. Different substrates may have different behaviors, so even though it inhibits glucuronidation by 40% in mouse gut for EGCG, it might be 80%, or 20%, for curcumin. It might be different in humans, too. Nevertheless, when you see something like this in one species, you look for it in another; it increases the likelihood that you will see it. I know that piperine has been used in humans for a number of compounds, and it increases their bioavailability. I don't know the split between gut and liver, or how much variation there is with substrates, but it clearly works. I think in all cases, the mechanism is inhibition of glucuronidation. In the mouse paper above, they also noted that it slowed intestinal transit time. I don't know if that's general behavior or not.

Swamping the gut with a second compound is an interesting idea. It's sort of the same as throwing an inhibitor in, as the second compound would be inhibiting the sulfation of the other by virtue of taking up space in the active site while it was being sulfated. It would be competing for sulfation capacity. Resveratrol is actually inhibiting its own sulfation when you get a high enough concentration. That's all that swamping is; just competitive self-inhibition, if you will.

The only problem I see with this and the train of thought it is following is the assumption that we don't want the sulfation and glucouronidation to occur. It seems that there are at least some who are familiar with these issues that indicate that the glucouoronidated form may be in that form specifically for transport to other organs who will in turn convert it back. If that is the case then swamping the gut may actually be obstructing a useful pathway rather than aiding it. Note I'm not familiar with this whole process this is just a summary of what I think I have read here.

Mike

Mike, this is one of those persistent myths that floats around imminst. Sulfation and glucuronidation are processes that have evolved to solubilize xenobiotics (molecules that originate outside the body) and effect their transport out of the body. That is their number one role; far and away that is all they do, and they do it very well. I'm not aware of any evidence that any of the conjugates (either glucuronide or sulfate) of resveratrol have sirtuin activating activity on their own. There are enzymes in the body that can cleave glucuronides, sulfates, esters, glycosides and other such linkages, and they seem to be widely expressed. However, when resveratrol is conjugated by one of these adducts, it is made substantially more hydrophilic. As such, it thermodynamically prefers to be in water rather than in a lipid membrane, so its ability to enter a cell is decreased by orders of magnitude. Unless a glycosidase or sulfatase activity is present in the blood or in some extracellular location, I can't see how conjugation is going to help resveratrol get into cells. It would be evolutionarily wrong for such a system to exist, as it would be undoing the very work that the body is doing to get rid of carcinogens and other toxins.

Some may be mislead by the existence of glucuronide prodrugs. I'm aware of at least one of these, and if I recall correctly it was done in order to circumvent a chemical instability. The fact that such a thing exists doesn't mean that glucuronidation helps all drugs get into cells, it means that in this special case, it was a method that could be exploited, and it may have required a huge increase in the dose in order to work. It is probably the case that a small fraction of circulating resveratrol glucuronide or sulfate makes its way into cells, but that might be one percent of what would get in if it were native resveratrol. Once it's in, it may get cleaved, which will assist it in leaving the cell.

One bit of evidence that conjugation is not helpful is the case of 4'O-acetyl resveratrol. In this compound, one of the sites of conjugation has been blocked by an acetyl group. In an in vitro experiment it was not a sirtuin activator, but in vivo it was substantially more effective than plain resveratrol. This is thought to occur because conjugation is blocked at the 4' position, so the acetylated compound was able to get into cells, whereupon an intracellular esterase cleaved the acetyl group, releasing free resveratrol. If conjugation was a good thing, you wouldn't see this effect.

Finally, I ran the idea that conjugation was a means of targeting drugs to intracellular sites of action past a professional pharmaceutical R&D person on the way to dinner tonight, as an example of goofy memes that float around here. She laughed. It's that ridiculous. I don't say that to be mean, because I know that you saw it here and you were just asking; I'm just trying to put a stake though the heart of this idea, which keeps popping up.

[docmaas]

How about some piperine to help with those gut enzymes?

It wouldn't hurt, (unless you have something else in your system that you'd really like to glucuronidate) but I don't know how much it will help. Sulfation is a bigger factor in taking resveratrol out, but glucuruonidation is also pretty significant. I don't know if piperine affects the gut in the same way as the liver or not.

By what method is piperine increasing curcumin blood levels?

edit. I think piperine does work in the gut.

What about swamping the gut with other substances that are sulphated, like curcumin.

http://jn.nutrition....full/134/8/1948

Piperine inhibits glucuronidation, generally speaking. From the paper you cite, it looks like for EGCG, it's effective in the gut in mice. In that model, it inhibits glucuronidation in the gut 40%, but it doesn't inhibit glucuronidation of EGCG in vitro with liver microsomes, which is pretty suggestive that it's not working in vivo in the liver, for EGCG in mice. Different substrates may have different behaviors, so even though it inhibits glucuronidation by 40% in mouse gut for EGCG, it might be 80%, or 20%, for curcumin. It might be different in humans, too. Nevertheless, when you see something like this in one species, you look for it in another; it increases the likelihood that you will see it. I know that piperine has been used in humans for a number of compounds, and it increases their bioavailability. I don't know the split between gut and liver, or how much variation there is with substrates, but it clearly works. I think in all cases, the mechanism is inhibition of glucuronidation. In the mouse paper above, they also noted that it slowed intestinal transit time. I don't know if that's general behavior or not.

Swamping the gut with a second compound is an interesting idea. It's sort of the same as throwing an inhibitor in, as the second compound would be inhibiting the sulfation of the other by virtue of taking up space in the active site while it was being sulfated. It would be competing for sulfation capacity. Resveratrol is actually inhibiting its own sulfation when you get a high enough concentration. That's all that swamping is; just competitive self-inhibition, if you will.

The only problem I see with this and the train of thought it is following is the assumption that we don't want the sulfation and glucouronidation to occur. It seems that there are at least some who are familiar with these issues that indicate that the glucouoronidated form may be in that form specifically for transport to other organs who will in turn convert it back. If that is the case then swamping the gut may actually be obstructing a useful pathway rather than aiding it. Note I'm not familiar with this whole process this is just a summary of what I think I have read here.

Mike

Mike, this is one of those persistent myths that floats around imminst. Sulfation and glucuronidation are processes that have evolved to solubilize xenobiotics (molecules that originate outside the body) and effect their transport out of the body. That is their number one role; far and away that is all they do, and they do it very well. I'm not aware of any evidence that any of the conjugates (either glucuronide or sulfate) of resveratrol have sirtuin activating activity on their own. There are enzymes in the body that can cleave glucuronides, sulfates, esters, glycosides and other such linkages, and they seem to be widely expressed. However, when resveratrol is conjugated by one of these adducts, it is made substantially more hydrophilic. As such, it thermodynamically prefers to be in water rather than in a lipid membrane, so its ability to enter a cell is decreased by orders of magnitude. Unless a glycosidase or sulfatase activity is present in the blood or in some extracellular location, I can't see how conjugation is going to help resveratrol get into cells. It would be evolutionarily wrong for such a system to exist, as it would be undoing the very work that the body is doing to get rid of carcinogens and other toxins.

Some may be mislead by the existence of glucuronide prodrugs. I'm aware of at least one of these, and if I recall correctly it was done in order to circumvent a chemical instability. The fact that such a thing exists doesn't mean that glucuronidation helps all drugs get into cells, it means that in this special case, it was a method that could be exploited, and it may have required a huge increase in the dose in order to work. It is probably the case that a small fraction of circulating resveratrol glucuronide or sulfate makes its way into cells, but that might be one percent of what would get in if it were native resveratrol. Once it's in, it may get cleaved, which will assist it in leaving the cell.

One bit of evidence that conjugation is not helpful is the case of 4'O-acetyl resveratrol. In this compound, one of the sites of conjugation has been blocked by an acetyl group. In an in vitro experiment it was not a sirtuin activator, but in vivo it was substantially more effective than plain resveratrol. This is thought to occur because conjugation is blocked at the 4' position, so the acetylated compound was able to get into cells, whereupon an intracellular esterase cleaved the acetyl group, releasing free resveratrol. If conjugation was a good thing, you wouldn't see this effect.

Finally, I ran the idea that conjugation was a means of targeting drugs to intracellular sites of action past a professional pharmaceutical R&D person on the way to dinner tonight, as an example of goofy memes that float around here. She laughed. It's that ridiculous. I don't say that to be mean, because I know that you saw it here and you were just asking; I'm just trying to put a stake though the heart of this idea, which keeps popping up.

I got all that and understand it. I am indeed only going by what others have suggested. My chemistry education was finishing just as biochemistry was emerging as a new discipline. I'm still reminded though of the folate/iron argument in the 50's I believe where the one person that in retrospect correctly indentified folate deficiency as a primary cause of what was assumed to be iron deficiency was overruled by "those who knew better". Only many years later was he proven to have been closer to the answer than those who had us eating out of cast iron pans and taking iron pills for our low hemoglobin for all those years. What you say makes sense but so have lots of other explanations that later turned out to be incorrect. There is yet another possibility and that is that the conjugants are actually active in entirely different but still related processes.

The 4'O-acetyl resveratrol is an interesting clue though. Have a look here: http://www.pubmedcen...mp;blobtype=pdf
I assume the 4'O molecule discussed here is the same as the one you mentioned. The in vitro experiment is very different but the results are very similar.

Is the acetyl bound resveratrol unstable or difficult to manufacture? I'm wondering if the bioavailability is greater because of blocked conjugation why not use it instead of straight resveratrol? The above article provides one way of methylating but it ends up with two different forms and I didn't see anything that indicated the ratios. It seems though from that paper that the methyl group is not as important as the location where it resides for its resisting breakdown in their research. Also pay attention to what they say when they cite reference (5).

Fascinating stuff.

Mike

[zoolander]

How can anyone make comments on bioavailability of micronized vs. non-micronized samples when it has been tested in humans? Perhaps I have missed a few things?

I saw this coming. Extrapolation from the results. Arguments between companies that stock resveratrol. I just find it hard to believe that an individual was prepared to run samples on HPLC and GC (that he happens to own) as well as taking photos of samples using a scanning electron microscope. The study became null and void when companies started sending hedgehog info samples. To avoid bias studies are required to be randomized and blinded (pref. double blinded). Hedgehog info receives labeled samples from companies and then runs the samples himself. It's very hard to avoid bias when that happens.

Yeah yeah I know. People are going to say that hedgehog info doesn't have any commercial interest in any of the companies involved but I remain a skeptic considering how the tests were conducted.

[niner]

How about some piperine to help with those gut enzymes?

It wouldn't hurt, (unless you have something else in your system that you'd really like to glucuronidate) but I don't know how much it will help. Sulfation is a bigger factor in taking resveratrol out, but glucuruonidation is also pretty significant. I don't know if piperine affects the gut in the same way as the liver or not.

By what method is piperine increasing curcumin blood levels?

edit. I think piperine does work in the gut.

What about swamping the gut with other substances that are sulphated, like curcumin.

http://jn.nutrition....full/134/8/1948

Piperine inhibits glucuronidation, generally speaking. From the paper you cite, it looks like for EGCG, it's effective in the gut in mice. In that model, it inhibits glucuronidation in the gut 40%, but it doesn't inhibit glucuronidation of EGCG in vitro with liver microsomes, which is pretty suggestive that it's not working in vivo in the liver, for EGCG in mice. Different substrates may have different behaviors, so even though it inhibits glucuronidation by 40% in mouse gut for EGCG, it might be 80%, or 20%, for curcumin. It might be different in humans, too. Nevertheless, when you see something like this in one species, you look for it in another; it increases the likelihood that you will see it. I know that piperine has been used in humans for a number of compounds, and it increases their bioavailability. I don't know the split between gut and liver, or how much variation there is with substrates, but it clearly works. I think in all cases, the mechanism is inhibition of glucuronidation. In the mouse paper above, they also noted that it slowed intestinal transit time. I don't know if that's general behavior or not.

Swamping the gut with a second compound is an interesting idea. It's sort of the same as throwing an inhibitor in, as the second compound would be inhibiting the sulfation of the other by virtue of taking up space in the active site while it was being sulfated. It would be competing for sulfation capacity. Resveratrol is actually inhibiting its own sulfation when you get a high enough concentration. That's all that swamping is; just competitive self-inhibition, if you will.

The only problem I see with this and the train of thought it is following is the assumption that we don't want the sulfation and glucouronidation to occur. It seems that there are at least some who are familiar with these issues that indicate that the glucouoronidated form may be in that form specifically for transport to other organs who will in turn convert it back. If that is the case then swamping the gut may actually be obstructing a useful pathway rather than aiding it. Note I'm not familiar with this whole process this is just a summary of what I think I have read here.

Mike

Mike, this is one of those persistent myths that floats around imminst. Sulfation and glucuronidation are processes that have evolved to solubilize xenobiotics (molecules that originate outside the body) and effect their transport out of the body. That is their number one role; far and away that is all they do, and they do it very well. I'm not aware of any evidence that any of the conjugates (either glucuronide or sulfate) of resveratrol have sirtuin activating activity on their own. There are enzymes in the body that can cleave glucuronides, sulfates, esters, glycosides and other such linkages, and they seem to be widely expressed. However, when resveratrol is conjugated by one of these adducts, it is made substantially more hydrophilic. As such, it thermodynamically prefers to be in water rather than in a lipid membrane, so its ability to enter a cell is decreased by orders of magnitude. Unless a glycosidase or sulfatase activity is present in the blood or in some extracellular location, I can't see how conjugation is going to help resveratrol get into cells. It would be evolutionarily wrong for such a system to exist, as it would be undoing the very work that the body is doing to get rid of carcinogens and other toxins.

Some may be mislead by the existence of glucuronide prodrugs. I'm aware of at least one of these, and if I recall correctly it was done in order to circumvent a chemical instability. The fact that such a thing exists doesn't mean that glucuronidation helps all drugs get into cells, it means that in this special case, it was a method that could be exploited, and it may have required a huge increase in the dose in order to work. It is probably the case that a small fraction of circulating resveratrol glucuronide or sulfate makes its way into cells, but that might be one percent of what would get in if it were native resveratrol. Once it's in, it may get cleaved, which will assist it in leaving the cell.

One bit of evidence that conjugation is not helpful is the case of 4'O-acetyl resveratrol. In this compound, one of the sites of conjugation has been blocked by an acetyl group. In an in vitro experiment it was not a sirtuin activator, but in vivo it was substantially more effective than plain resveratrol. This is thought to occur because conjugation is blocked at the 4' position, so the acetylated compound was able to get into cells, whereupon an intracellular esterase cleaved the acetyl group, releasing free resveratrol. If conjugation was a good thing, you wouldn't see this effect.

Finally, I ran the idea that conjugation was a means of targeting drugs to intracellular sites of action past a professional pharmaceutical R&D person on the way to dinner tonight, as an example of goofy memes that float around here. She laughed. It's that ridiculous. I don't say that to be mean, because I know that you saw it here and you were just asking; I'm just trying to put a stake though the heart of this idea, which keeps popping up.

I got all that and understand it. I am indeed only going by what others have suggested. My chemistry education was finishing just as biochemistry was emerging as a new discipline. I'm still reminded though of the folate/iron argument in the 50's I believe where the one person that in retrospect correctly indentified folate deficiency as a primary cause of what was assumed to be iron deficiency was overruled by "those who knew better". Only many years later was he proven to have been closer to the answer than those who had us eating out of cast iron pans and taking iron pills for our low hemoglobin for all those years. What you say makes sense but so have lots of other explanations that later turned out to be incorrect. There is yet another possibility and that is that the conjugants are actually active in entirely different but still related processes.

The 4'O-acetyl resveratrol is an interesting clue though. Have a look here: http://www.pubmedcen...mp;blobtype=pdf
I assume the 4'O molecule discussed here is the same as the one you mentioned. The in vitro experiment is very different but the results are very similar.

Is the acetyl bound resveratrol unstable or difficult to manufacture? I'm wondering if the bioavailability is greater because of blocked conjugation why not use it instead of straight resveratrol? The above article provides one way of methylating but it ends up with two different forms and I didn't see anything that indicated the ratios. It seems though from that paper that the methyl group is not as important as the location where it resides for its resisting breakdown in their research. Also pay attention to what they say when they cite reference (5).

Well, this is pretty well trod ground, so in this case I think those who know better are probably on to something. The molecule in the paper you cite was a methyl ether which would be relatively stable metabolically. The acetyl version can be cleaved by intracellular esterases, so it's pretty interesting. It's not that hard to make. I think the problem with it boils down to it not falling under the "supplement" definitions; if it's a synthetic product not found in nature, then it's treated like a new drug. I believe there's an enzymatic synthesis in the literature that starts with resveratrol. You could shotgun acetylate it, but then you'd have to separate the various isomers. I don't have access to the full text of reference 5 (Youdim et al.), but it sounded pretty speculative. Youdim's comments may have been more directed to other phytochemicals than resveratrol, I don't know. In the six years since that review came out, there has been a lot of work on resveratrol pharmacokinetics. Everything I've seen leads me to believe that conjugation is taking resveratrol out of the picture rather than helping deliver it. It's not inconceivable that resveratrol metabolites could sneak into cells at a low level, but it would be a minor pathway at best. If it really worked well, mammals wouldn't need huge doses of resveratrol to see the effects we're seeing, because the blood levels of the metabolites are certainly sky high.

[docmaas]

How can anyone make comments on bioavailability of micronized vs. non-micronized samples when it has been tested in humans? Perhaps I have missed a few things?
Well for starters you missed the "n't" in the word "has" in the sentence above.
As a few have pointed out particle size diminution and increased activity are pretty commonly associated. It may be a generalization but given the way these things more often than not work it is not a very risky one IMHO.

I saw this coming. Extrapolation from the results.
There is a difference between extrapolation and hypothesis.
Arguments between companies that stock resveratrol.
I must have missed this. The only person from a company involved in this discussion is Anthony.

I just find it hard to believe that an individual was prepared to run samples on HPLC and GC (that he happens to own) as well as taking photos of samples using a scanning electron microscope.
I think you've failed to complete your sentence or thought. Why would he have the equipment if he weren't willing and able to use it?

The study became null and void when companies started sending hedgehog info samples.
That is certainly problematic. On the other hand if he received them in factory packages the likelihood of their having been inauthentic would go down somewhat. Of course it would have been better had they been purchased without the knowledge of the provider as to their purpose. But this isn't an institution funded by a government grant it's a collection of interested people some of whom have more knowledge and resources than others.

If hedge wants to do more research which I find of great benefit I would certainly be happy to contribute monetarily to his efforts so he could provide for things like purchases from retail sources and maybe pay for some of the materials he is using.

To avoid bias studies are required to be randomized and blinded (pref. double blinded). Hedgehog info receives labeled samples from companies and then runs the samples himself. It's very hard to avoid bias when that happens.
Test results are facts not judgements. There is always room for uconscious bias in the examination of the results and the interpretation of what they might mean but the purity test and the pictures were published so we could all see them. They could have been falsified but you really should posit a reason for the possibility they were otherwise it is hard to believe why someone would do so.

Yeah yeah I know. People are going to say that hedgehog info doesn't have any commercial interest in any of the companies involved but I remain a skeptic considering how the tests were conducted.

Well frankly I'm more inclined to think you have some kind of hidden motive here rather than Hedge. Otherwise why would you come up with this kind of stuff?

Mike

Edited by docmaas, 10 February 2008 - 09:06 AM.

[zoolander]

As a few have pointed out particle size diminution and increased activity are pretty commonly associated. It may be a generalization but given the way these things more often than not work it is not a very risky one IMHO

Correct me if I am wrong but I thought the bioavailability issues with resveratrol are related to glucoronidation.

There is a difference between extrapolation and hypothesis.

Perhaps extrapolation is not the right word then. Maybe I should have use the work "suggestion". I don't think the following is a hypothesis though

NINER: Hedgehog, thanks for doing all of this! Is the Trans-Max the version that's supposed to have the secret bioavailability enhancement? If so, it would appear to be micronization. If not, well, it looks like it was micronized. Micronization really should result in better absorption for a solubility challenged compound like this. I think Biotivia is claiming a factor of two for the secret process. I've heard various versions of Sirtris' claim, some as high as a factor of ten, but after the recent human trials I heard a factor of five. But SRT-501 has more going on than micronization alone.


HEDGEHOG: Yes micronization does help in the bioavailability. How much? A lot of companies need micronization for manufacturing purposes. Lets say you have a 5mg tablet.... How the heck are you going to make a 5mg tablet if you have large particles or clumps. you need fine powder todo this.

To be honest both of these samples have small particle size. This is probably due to the extraction process. Many drugs have much larger particle size.

I must have missed this. The only person from a company involved in this discussion is Anthony.

In this discussion perhaps but there is something going on on yahoo between Anthony and Mr Biotiva

That is certainly problematic. On the other hand if he received them in factory packages the likelihood of their having been inauthentic would go down somewhat. Of course it would have been better had they been purchased without the knowledge of the provider as to their purpose. But this isn't an institution funded by a government grant it's a collection of interested people some of whom have more knowledge and resources than others.

A separate party could have been used to blind the study.

Test results are facts not judgements. There is always room for uconscious bias in the examination of the results and the interpretation of what they might mean but the purity test and the pictures were published so we could all see them. They could have been falsified but you really should posit a reason for the possibility they were otherwise it is hard to believe why someone would do so.

tests results can become judgements and that why it's best to remove the possibilty of this by randomising and blinding the samples.

Well frankly I'm more inclined to think you have some kind of hidden motive here rather than Hedge. Otherwise why would you come up with this kind of stuff?

My motive is good science. My side motive is also to point out how dubious such studies can be. It's very difficult to expose conflicts of interest especially in a forum like this one. So I'll play the devils advocate. Perhaps I should just let you guys do what you do and ignore this thread.

[Anthony_Loera]

Ok, I see your point Zoolander.

Docmass, remember Hedge said that James sent him 30 capsules.
I maybe the only one posting, but I am not the only one reading.

A

[Hedgehog]

Ok, I see your point Zoolander.

Docmass, remember Hedge said that James sent him 30 capsules.
I maybe the only one posting, but I am not the only one reading.

A

Ok, I personally bought some Rev99 form Anthony (I did this online w/o talking to him) before I even stated I was going to conduct the tests. He didn't know about this. I was sent one capsule of Trans-Max from a imminist member. I did the purity test on this sample. The SEM results and uniformity results are from a undamaged and sealed packed that came from Jame's stockpile. Like I said the uniformity results SUCKED for all vendors. But I guess they sucked in a good way! As for the SEM results. Yes I used the sent samples from the vendor. If somebody else wants to send me one more trans-max pill to make it more unbiased feel free to. However, I will need it sent to me by the end of next week.

[docmaas]

As a few have pointed out particle size diminution and increased activity are pretty commonly associated. It may be a generalization but given the way these things more often than not work it is not a very risky one IMHO

Correct me if I am wrong but I thought the bioavailability issues with resveratrol are related to glucoronidation.

They are but not exclusively. The first place it will come into play is in the gut if taken orally. The smaller the particles the more opportunities there are for reactions between the particles and other chemicals in the gut. It's just a numbers game. If you take a one gram chunk and it doesn't break down any smaller you'll have just the surface area of the large chunk with which to react. If you break that 1 gram chunk in 1000 1 mg chunks you'll have a lot more surface area and if it broken down into even smaller chunks even more surface area. If you put a large piece of crystalline sugar into water and a teaspoon of granulated sugar into water both the same quantity which one will dissolve faster? Ever try making coffee without grinding the beans? Does coarse ground coffee slow water down in the filter as much as fine ground coffee and is the coffee as strong when made with a coarse grind as with a fine grind?

There is a difference between extrapolation and hypothesis.

Perhaps extrapolation is not the right word then. Maybe I should have use the work "suggestion". I don't think the following is a hypothesis though

NINER: Hedgehog, thanks for doing all of this! Is the Trans-Max the version that's supposed to have the secret bioavailability enhancement? If so, it would appear to be micronization. If not, well, it looks like it was micronized. Micronization really should result in better absorption for a solubility challenged compound like this. I think Biotivia is claiming a factor of two for the secret process. I've heard various versions of Sirtris' claim, some as high as a factor of ten, but after the recent human trials I heard a factor of five. But SRT-501 has more going on than micronization alone.

It's a hypothesis or a suggestion; a distinction without a difference.


HEDGEHOG: Yes micronization does help in the bioavailability. How much? A lot of companies need micronization for manufacturing purposes. Lets say you have a 5mg tablet.... How the heck are you going to make a 5mg tablet if you have large particles or clumps. you need fine powder todo this.

To be honest both of these samples have small particle size. This is probably due to the extraction process. Many drugs have much larger particle size.

I must have missed this. The only person from a company involved in this discussion is Anthony.

In this discussion perhaps but there is something going on on yahoo between Anthony and Mr Biotiva.

The point is that in this discussion only Anthony is here not James and as far as I can tell there is no one here taking the position of being an advocate for Biotivia.

That is certainly problematic. On the other hand if he received them in factory packages the likelihood of their having been inauthentic would go down somewhat. Of course it would have been better had they been purchased without the knowledge of the provider as to their purpose. But this isn't an institution funded by a government grant it's a collection of interested people some of whom have more knowledge and resources than others.

A separate party could have been used to blind the study.

That's true and in an academic or research paradigm that would have been done but it takes a whole different kind of organization than an internet forum to provide the resources and structure for those kinds of studies. Moreover, I personally would still need either a reason for or evidence of hidden motivation on the part of Hedge or any other respondent in order to doubt their good intentions or their veracity.

Test results are facts not judgements. There is always room for uconscious bias in the examination of the results and the interpretation of what they might mean but the purity test and the pictures were published so we could all see them. They could have been falsified but you really should posit a reason for the possibility they were otherwise it is hard to believe why someone would do so.

tests results can become judgements and that why it's best to remove the possibilty of this by randomising and blinding the samples.
But the test results were published in a form in which each of us could make our own judgements. The only place bias could have entered here was in the selection of which pictures to present. I'm willing to accept and assume that such a decision was made on a basis of presenting the most usable photos rather than those that favored one product over the other until I have evidence of some other motivation.

Randomizing and blinding could have been done but it really isn't necessary in this situation IMHO. I will send him a sample from the transmax I purchased from Biotivia before I ever joined this group.

Well frankly I'm more inclined to think you have some kind of hidden motive here rather than Hedge. Otherwise why would you come up with this kind of stuff?

My motive is good science. My side motive is also to point out how dubious such studies can be. It's very difficult to expose conflicts of interest especially in a forum like this one. So I'll play the devils advocate. Perhaps I should just let you guys do what you do and ignore this thread.

I understand your concerns. I just don't agree with them. But I encourage you to continue to voice them. In fact I probably am just as far out in the opposite direction to you in terms of skepticism in this particular situation. Stick around we'll counterbalance each other.

I think a good example of scientific skepticism though is Niner's approach to my hypotheses about the conjugates of resveratrol. He is not doubting me but rather my ideas which is healthy and he is doing it based on much more prior experience than I have. I too am just playing devil's advocate because as everyone knows who does science hypotheses have to change to account for new information and sometimes that information comes much later and often in surprising ways from surprising directions.

Mike

[docmaas]

Hedge,

I'll send you out a capsule tomorrow. You should get it on Wednesday. I'm down in Irvine, CA right now.

Mike

Ok, I see your point Zoolander.

Docmass, remember Hedge said that James sent him 30 capsules.
I maybe the only one posting, but I am not the only one reading.

A

Ok, I personally bought some Rev99 form Anthony (I did this online w/o talking to him) before I even stated I was going to conduct the tests. He didn't know about this. I was sent one capsule of Trans-Max from a imminist member. I did the purity test on this sample. The SEM results and uniformity results are from a undamaged and sealed packed that came from Jame's stockpile. Like I said the uniformity results SUCKED for all vendors. But I guess they sucked in a good way! As for the SEM results. Yes I used the sent samples from the vendor. If somebody else wants to send me one more trans-max pill to make it more unbiased feel free to. However, I will need it sent to me by the end of next week.

[docmaas]

Hi Anthony,

I will send Hedge a capsule of transmax I got in an order made back in December. If the Sem photos show the same granulation as those he got from James that should put this one to bed. If not then something is amiss.

Mike

Ok, I see your point Zoolander.

Docmass, remember Hedge said that James sent him 30 capsules.
I maybe the only one posting, but I am not the only one reading.

A

[Hedgehog]

How can anyone make comments on bioavailability of micronized vs. non-micronized samples when it has been tested in humans? Perhaps I have missed a few things?

I saw this coming. Extrapolation from the results. Arguments between companies that stock resveratrol. I just find it hard to believe that an individual was prepared to run samples on HPLC and GC (that he happens to own) as well as taking photos of samples using a scanning electron microscope. The study became null and void when companies started sending hedgehog info samples. To avoid bias studies are required to be randomized and blinded (pref. double blinded). Hedgehog info receives labeled samples from companies and then runs the samples himself. It's very hard to avoid bias when that happens.

Yeah yeah I know. People are going to say that hedgehog info doesn't have any commercial interest in any of the companies involved but I remain a skeptic considering how the tests were conducted.

I understand zoolander. If I do this again next I will have ppl just send me samples and won't ask where they came from. Rather they can disclose this information to the forum after I post the results? However, I didn't think of this nor did I think it was a problem. I can give you the protocol that I followed (based on a published study).

I have already stated that I'm working on a formulation. Personally I need to see if the methods of testing actually work. In other words I don't want to test my blood plasma and then figure out that the method doesn't work. That would really really suck for me.

All the Resveratrol vendors have a good product. If I was paid by Anthony or James or another vendor the results could have been skewed. IMO I think the results had good consistency meaning there is little difference between vendors and methods worked pretty well. They also had a very similar impurity profile. In a personal communication I was told there are only a few large scale manufactures of Resveratrol. If you look at the 50X SEM results you can see the complete sample that I used. As for zooming in. Yes that was biased. It is biased for anybody. Where do I want to zoom in? Well I moved the sample around until I got what I felt like was a good image and representation of sample. Download the 50-100x image and zoom in on your computer. That works pretty well and you won't have to rely on my zooming…

Warmest Regards,

~Hh

My results also matched up very well with resveratrol vendors CoA. Which gives me confidance that the methods worked or at least matched up with other lab results.

Edited by hedgehog_info, 11 February 2008 - 12:44 AM.

[zoolander]

How about I assign another party from this forum. The vendors will send their samples to this person. This person will label (blind) the samples and then send the samples to you. You then run the sample on HPLC and GC as you have already done in triplicut. Take the SEM photos and then publish the results here with the assigned labels. These labels will then be adjusted to the original sample names by the person who blinded the study.

Also, have your HPLC and GC methods been validated by more than one lab? Is the method a published method? If so can you please provide the reference?

You will need to add in a positive control. For example you need to purchase some analytical grade resveratrol from sigma aldrich

Resveratrol
Empirical Formula (Hill Notation): C14H12O3, Formula Weight: 228.24, CAS Number: 501-36-0
R5010 >99% (GC) (Sigma)
34092 VETRANAL®, analytical standard (Riedel-de Haën)

This also needs to be sent to the assigned individual

[bixbyte]

How about I assign another party from this forum. The vendors will send their samples to this person. This person will label (blind) the samples and then send the samples to you. You then run the sample on HPLC and GC as you have already done in triplicut. Take the SEM photos and then publish the results here with the assigned labels. These labels will then be adjusted to the original sample names by the person who blinded the study.....

Dear mr Zoolander,

I mailed Hh the vital prime 99% Resveratrol sample to test so I would know if their claim for 99 was true.
If I had not mentioned this, I would be anonymous.
What is your point?
Double Blind HPLC GC res tests on a BB for Res users that share info.
I think you are going too far.
Did Hh do anything wrong?

Bix

[zoolander]

It wouldn't take much to increase the power of the study being conducted. If you're going to do the study why not do it right?

[maxwatt]

How about I assign another party from this forum. The vendors will send their samples to this person. This person will label (blind) the samples and then send the samples to you. You then run the sample on HPLC and GC as you have already done in triplicut. Take the SEM photos and then publish the results here with the assigned labels. These labels will then be adjusted to the original sample names by the person who blinded the study.....

Dear mr Zoolander,

I mailed Hh the vital prime 99% Resveratrol sample to test so I would know if their claim for 99 was true.
If I had not mentioned this, I would be anonymous.
What is your point?
Double Blind HPLC GC res tests on a BB for Res users that share info.
I think you are going too far.
Did Hh do anything wrong?

Bix

I too believe that under the circumstances, there would be little to gain by this exacting procedure. Hh is not a resveratrol vendor, at least not yet, and I do not believe he has anything to gain by biasing the results. The tests may not be up to research grade standards, but I believe they are up to forensic standards.

The tests have only shown that a res is a res is a res.

[niner]

How about I assign another party from this forum. The vendors will send their samples to this person. This person will label (blind) the samples and then send the samples to you. You then run the sample on HPLC and GC as you have already done in triplicut. Take the SEM photos and then publish the results here with the assigned labels. These labels will then be adjusted to the original sample names by the person who blinded the study.

If we were going to do this, we certainly couldn't let the vendors know that the sample was going to be analyzed. It would have to be purchased anonymously on the open market. It's also not entirely fair to the vendor if we use a sample from an individual who may have bought it a long time ago and not stored it properly. At least those are all things to think about if we were going to do this again. Really, given the results that hedgehog got, (everyone looked pretty good) I'm not sure I see much of a need for doing all the work over again.

As hedgehog said, he's working on some sort of new resveratrol formulation, and I'd kind of like to see what he comes up with, since he seems to have the tools to do it right.

[Hedgehog]

It wouldn't take much to increase the power of the study being conducted. If you're going to do the study why not do it right?

Hi Zoolander,

To be honest I think sigmas purity GC test sux (a better way is HPLC). They have to chemically modify resveratrol (TMS) in order to test its purity by GC. Do you think that is a good idea. I think it just adds another dimension. Yes if you want to get VERY quick results then use a GC for purity. But can this detect cis/tran? I would be very interested to know if a GC method could detect both isomers.
Sigmas CoA includes GC and TLC. Many vendors do many other analytical tests compared to Sigma. Sure sigma carries a big name but that doesn't mean it is the best?

If I was going to sell my service or release a CoA for somebody then I might take your thoughts into consideration. But there is no reason to treat me being a analyst as a clinical trial. If you want to pay a lab to do this then go for it.

So far my data has matched up well with third party vendors CoA's. So you can take my experimentation results or forget about them. It would be wonderful if you could get somebody to see if they get similar images/results. Heck we could even do it blinded this time to make sure they are not biased.

Regards,
~Hh

GC Derivatizing Agents
Why is chemical derivatization needed?
Gas chromatography, a technique for separation of volatile compounds which are thermally stable, is unfortunately not always easily to apply for compounds of biomedical and environmental interest, particularly for those of high molecular weight or containing polar functional groups. These groups are difficult to analyze by GC either because they are not sufficiently volatile, tail badly, are too strongly attracted to the stationary phase, thermally unstable or even decomposed.


http://www.sigmaaldr...ing_Agents.html

Edited by hedgehog_info, 11 February 2008 - 06:08 AM.

[docmaas]

It wouldn't take much to increase the power of the study being conducted. If you're going to do the study why not do it right?

"I could see this coming" to paraphrase your initial post. This isn't an academic or commercial enterprise. Why not be a bit more trusting? How can we know that you won't substitute your own materials for those that are sent to you? No matter how many safeguards you put in place there are always places that things can be altered if someone has the intent to do so.

I'm just fine with the way things are being done now. I don't believe Hedge has an axe to grind and I am quite uncomfortable with your lack of trust if not in him in the process.

Mike

[zoolander]

Is there any reason why you continue to use red when you respond to my posts docmaas? I would appreciate it if you stuck to the one color font. The red comes accross as you yelling/angry.

I'm just fine with the way things are being done now. I don't believe Hedge has an axe to grind and I am quite uncomfortable with your lack of trust if not in him in the process.

We'll have to agree to disagree then Mike. As a scientist I have my views.

I am quite uncomfortable with your lack of trust if not in him in the process

Care factor = Zero. Harden up Mike.

Now please drop the red font or I will.

Edited by zoolander, 11 February 2008 - 06:59 AM.

[Hedgehog]

More images... I also got a Trans-Max pill from Docmass. You can also download them from here http://www.pathway2c...SEM/New Folder/

China PC-091226 100x


China PC-091226 250x

China PCE-080116 100



China PCE-080116 250



T-Max 100x




T-Max 250

[edward]

Hedgehog,

Any way you can put some Vital Prime under that electron microscope?

[Hedgehog]

Hedgehog,

Any way you can put some Vital Prime under that electron microscope?

Only for you.

It might take me a while... I need to find the sent sample. Bix did you send me the VitalPrime sample?

[edward]

Thanks

[Hedgehog]

Thanks

Hey Ed do you have some VP product? It is very strange it is like a mix of all the other vendors. It might even be finer that T-Max, but is has some LARGE clumps. I think some of those clumps are from the static and the small pieces coming together. VP stuff was hard to get off my spatula probably becuase it was sent in a plastic bag. I also noticed some larger crystals like seen in other vendors. I will upload the pics soon. Very fun to look at, i took many pics and went all the way up to 1500x because I honestly didn't really understand what I was looking at.

For those of you who have used VP, does some of it dissolve fast and some doesn't dissolve at all?

Edited by hedgehog_info, 21 February 2008 - 04:32 PM.

[edward]

Thanks

Hey Ed do you have some VP product? It is very strange it is like a mix of all the other vendors. It might even be finer that T-Max, but is has some LARGE clumps. I think some of those clumps are from the static and the small pieces coming together. VP stuff was hard to get off my spatula probably becuase it was sent in a plastic bag. I also noticed some larger crystals like seen in other vendors. I will upload the pics soon. Very fun to look at, i took many pics and went all the way up to 1500x because I honestly didn't really understand what I was looking at.

For those of you who have used VP, does some of it dissolve fast and some doesn't dissolve at all?

Its funny you mention these things as they are consistent with what I have observed.

What I have noticed is that it "packs" much more tightly into my 1/4 teaspoon measuring spoon so 1/4 teaspoon when weighed ends up being around 700-800 mg versus 500-600 mg which was how much the average 1/4 teaspoon of my last Mega shipment weighed. Yes, it does tend to stick to the spoon more than other 98 and 99 products I have had. I assumed that both of these characteristics were because of the fact that the product wasn't dried thoroughly as some have speculated and contained some moisture (confirmed as 2% of the powder by weight by the lab). It would be great though if it was because the product was finer and thus denser.

In warm water with Miralax (PEG), some of it does disperse/dissolve very quickly with simple stirring (by my perception quicker then my Mega 99%) but there is also more at the bottom of the container that doesn't dissolve when compared to Mega, I can get more to dissolve/disperse with a hand blender, breaking up the clumps? but not as much as with Mega. All of it dissolves just fine in in oil, takes a little stirring and breaking up of some clumps but not bad (mixture of fish oil and warmed coconut oil). I haven't tried EtOH with the Vital Prime as I decided that it was not a viable option for me to be taking shots of vodka or everclear each morning

edit: anxiously awaiting pictures

Edited by edward, 21 February 2008 - 05:51 PM.

[Hedgehog]

Thanks

Hey Ed do you have some VP product? It is very strange it is like a mix of all the other vendors. It might even be finer that T-Max, but is has some LARGE clumps. I think some of those clumps are from the static and the small pieces coming together. VP stuff was hard to get off my spatula probably becuase it was sent in a plastic bag. I also noticed some larger crystals like seen in other vendors. I will upload the pics soon. Very fun to look at, i took many pics and went all the way up to 1500x because I honestly didn't really understand what I was looking at.

For those of you who have used VP, does some of it dissolve fast and some doesn't dissolve at all?

Its funny you mention these things as they are consistent with what I have observed.

What I have noticed is that it "packs" much more tightly into my 1/4 teaspoon measuring spoon so 1/4 teaspoon when weighed ends up being around 700-800 mg versus 500-600 mg which was how much the average 1/4 teaspoon of my last Mega shipment weighed. Yes, it does tend to stick to the spoon more than other 98 and 99 products I have had. I assumed that both of these characteristics were because of the fact that the product wasn't dried thoroughly as some have speculated and contained some moisture (confirmed as 2% of the powder by weight by the lab). It would be great though if it was because the product was finer and thus denser.

In warm water with Miralax (PEG), some of it does disperse/dissolve very quickly with simple stirring (by my perception quicker then my Mega 99%) but there is also more at the bottom of the container that doesn't dissolve when compared to Mega, I can get more to dissolve/disperse with a hand blender, breaking up the clumps? but not as much as with Mega. All of it dissolves just fine in in oil, takes a little stirring and breaking up of some clumps but not bad (mixture of fish oil and warmed coconut oil). I haven't tried EtOH with the Vital Prime as I decided that it was not a viable option for me to be taking shots of vodka or everclear each morning

edit: anxiously awaiting pictures

Will I would need to actually brake those clumps apart to see what they are made of. I can't really do that easily. My guess is that there are larger particles underneath them that are attracting the very fine resveratrol particles.

I have to process some data before I can post images... I will keep you anxiously waiting

[Hedgehog]

http://www.pathway2c...eratrol/SEM/VP/

If somebody wants to repost the images on the forum go for it.

[Hedgehog]

http://www.pathway2c...eratrol/SEM/VP/

If somebody wants to repost the images on the forum go for it.

As you can tell there are a lot of clumps but they don't look like large cyrstals rather a lot of small particles smashed together. There are a few larger particles and you can see that the surface looks much different.

Very interesting.

Edited by hedgehog_info, 21 February 2008 - 08:44 PM.