Optimal AHK-Cu Proportioning: Math Inside.

[TheLastHairbender]

So I know there has been a good deal of talk about using AHK-Cu lately, and a number of people seem to be happily beta testing various combinations of the pure stuff from last autumn. I'm one of those - sitting on 28g right now but have not yet taken the initiative to work out the recipe for optimal application.

The best treatment of the topic I've seen to date are: mix a few grams into a bottle of Tricomin or ethanol/PG/H20. I'm a little hesitant to start experimenting based on such imprecise chemistry. I wanted to take a look at the original study and determine exactly how many grams need to be mixed into an amount of the vehicle appropriate for daily application. Most advice on the topic seems to center around mixing a 2.5% - 5% w/v solution of AHK-Cu, but without any mention of the volume you should be applying. I think the right approach is to determine how many moles of AHK-Cu you want to deliver and from there determine the amount (in weight) of AHK-Cu required to provide those moles, from which you can solve for the strength of solution you'd need for a given volume of solution you plan to apply.



Looking at the Korean study, they treated cells with solutions of varying concentrations, from 10^-12 M to 10^-7 M. They found positive results for concentrations from 10^-12 M to 10^-9 M, and negative results for concentrations of 10^-8 M and 10^-7 M (not sure how tenuous the differences between those amounts are...factors of ten for each increment to the exponent, right? So those are pretty discrete intervals if I understand correctly). The best results were obtained for a concentration of 10^-9 M.

I plan to apply 2 mL of solution daily and want to determine the number of grams of a compound I need to add to achieve the desired quantity in moles. The molecular weight of the compound appears to be 416.94u.

I thought I just needed to multiply the number of moles desired, 1.0 x 10^-9, by the molar mass, 416.94 g/mol, to determine the mass, in grams, required to deliver said moles. This works out to a tiny amount... 4.41694 x 10^-7 (.00000041694) grams per day. This seems way too small. Maybe I actually want to deliver more moles than that...? The study was on units of 30 follicular dermal papilla cells...so maybe I need to multiply the number of moles they delivered to their sample by the number of samples I'd expect to have on my scalp…about ~30,000 samples? I'm out of my league here and want to make sure I do this properly if I do it at all...unfortunately the lack of precedent in this area and the one study available (in which the treatment process doesn't seem to be well described) doesn't give me much confidence that I'm doing this right. There's actually a good chance I'll end up saying this isn't a good idea, but I wanted to give it a go to see if what I'm after can be scientifically determined with relative ease.

People who have attacked this less scientifically report adding approximately 2g to 60 mL (57g) of solution, which yields a 3.5% w/v concentration, and if 2 mL are applied daily yields a mass of active ingredient of .067 grams applied daily. Way higher than the 4.1694 x 10^-7 grams per day I solved for based on the number of moles I wanted to deliver. Which is the right way to think about this? I think the problem is that I don't exactly know how much I need to deliver...I would think the study should reveal that information, in g/kg or some equivalent that might be useful. Here is a link to to the study in question, maybe that quantity jumps out at someone more than it does to me:*http://www.dermoday.com/dosyalar/1277308389.pdf

If anyone knows their stoichiometry please advise. If you're just going to say make a 2.5% w/v solution and not tell me how much volume that assumes you're applying then I'm going to know you don't know what you're talking about, as 1 mL of a 2.5% solution is a lot different than 4 mL of 2.5% solution. I'd really prefer to hear from someone who understands the molarity calculations, if such a person exists on HairLossTalk.com. I've asked a few other people who might be in the know from a chemistry perspective; whether they're interested in staking my health on their back-of-the envelope calculations I'm not sure yet, but I'll post back whatever I hear. I appreciate anyone's input in the meantime.

 

[Jacob]

I still don't get why they(Procyte) didn't go with that % if it worked so well. Even if it's the cost..they could have had a second separate product that many would have been willing to pay for.

 

[Bryan]

The best treatment of the topic I've seen to date are: mix a few grams into a bottle of Tricomin or ethanol/PG/H20. I'm a little hesitant to start experimenting based on such imprecise chemistry. I wanted to take a look at the original study and determine exactly how many grams need to be mixed into an amount of the vehicle appropriate for daily application. Most advice on the topic seems to center around mixing a 2.5% - 5% w/v solution of AHK-Cu, but without any mention of the volume you should be applying. I think the right approach is to determine how many moles of AHK-Cu you want to deliver and from there determine the amount (in weight) of AHK-Cu required to provide those moles, from which you can solve for the strength of solution you'd need for a given volume of solution you plan to apply.

Forget about the "moles" you want to deliver, just worry about the total weight in grams of the copper peptide that you want to apply. That will give you a general idea of how much of a given solution you need to apply.

Looking at the Korean study, they treated cells with solutions of varying concentrations, from 10^-12 M to 10^-7 M...

What "Korean study" are you talking about?? :dunno:

 

[armandein]

The effect of tripeptide-Cooper complex on human hair-growth in vitro

http://www.dermoday.com/dosyalar/1277308389.pdf

Did you read the post?

 

[Bryan]

No, I've never seen or heard of that study before.

 

[TheLastHairbender]

The Korean study I referred to was the one from Seoul National University's Dermatology department, linked to in the original post:

http://www.dermoday.com/dosyalar/1277308389.pdf


I understand your saying to forget the quantity in moles and consider only the quantity in grams. Since one is just a linear transformation of the other - a multiplication or division by the substance's molar mass - the two measures are equivalent. (I.E. to deliver ten times more moles you need to deliver ten times more grams, for example). So I agree, knowing the quantity to deliver in grams is sufficient.

I started with the number of moles, though, because that was the only measured treatment quantity provided by existing research. I've never seen anything more than an anecdote that suggests the appropriate mass of AHK-Cu to deliver in grams...does anyone have information about this?

 

[Bryan]

I understand your saying to forget the quantity in moles and consider only the quantity in grams. Since one is just a linear transformation of the other - a multiplication or division by the substance's molar mass - the two measures are equivalent. (I.E. to deliver ten times more moles you need to deliver ten times more grams, for example). So I agree, knowing the quantity to deliver in grams is sufficient.

I started with the number of moles, though, because that was the only measured treatment quantity provided by existing research. I've never seen anything more than an anecdote that suggests the appropriate mass of AHK-Cu to deliver in grams...does anyone have information about this?

Rather than just use that Korean study, I'd suggest also using the Trachy et al studies which I've posted several times on hairloss sites. They aren't as highly technical as the Korean study, but they do give some details of a larger number of copper peptides than just the one used in Tricomin. Furthermore, these studies used more than just one kind of animal with which to experiment, including humans. The human trial used what they call "PC1031", not the "PC1234" used in Tricomin. It was applied in both 2% and 10% solutions of the peptide in propylene glycol, alcohol, hydroxypropylmethylcellulose, water, and nonoxynol 9. Application of 0.25 mL was made morning and evening by each subject.

PC1234 and PC1031 were also applied topically in a separate study, but only in mice. The PC1234 (which was more effective than the PC1031) was applied in doses of 0.25%, 0.67%, and 1.25%, while the PC1031 was applied in doses of 1.0%, 2.5%, 5.0%. The PC1234 definitely had better results, even at those lower doses.

For more details about all three of these studies, obtain them from a medical library, and read them in their entirety (I've given the full citations in previous posts).

 

[TheLastHairbender]

Ok, thanks for the references.

In the human study, they applied .25 mL (just a couple drops really) of solution, which would be about .237 grams of solution, so the 2% strength would contain .0047g (4.7mg) of AHK-Cu per application and the 10% would contain .0237g (23.7mg) of AHK-Cu per application. That sounds in line with what people have been suggesting, and sounds like a reasonable scaling up of the amounts used in the mouse studies in the Trachy et al patents: .36mg to .55mg of copper-peptide per .1mL injection.

At those amounts, twice daily applications (.25mL each) at 2% would require only .28 grams of AHK-Cu per month, while two daily applications at 10% would need 1.4 grams of AHK-Cu per month, dissolved in 15mL of solution. Sounds reasonable.


If I were to go on that knowledge, and believing that 1 mL (~.95g) of solution per application would probably be ideal for coverage of my particular balding pattern, to deliver the maximum treatment group's 23.7mg per application I would wind up with a 2.5% strength solution ( .0237g / .95 g ) of which I apply 1 mL twice daily. So for a monthly batch I would mix 1.42g of AHK-Cu into 60 mL of solution. That's exactly what I wanted to know. I'll be using 23.7mg per application, which is 47.4mg/day.

It might be easier to administer double the dose and apply it once a day, but since 47.4mg per application is outside the known treatment range for a single dose, I'd rather not be a guinea pig for the effects of that quantity of AHK-Cu at one time. Instead I'll bear the inconvenience for safety's sake and use 1 mL at 2.5% strength twice daily.

I've heard about possible interference with minoxidil cited as a reason for not simply mixing the appropriate quantity of AHK-Cu into a bottle of liquid minoxidil, so I plan to still apply liquid minoxidil first thing in the morning, followed in 15-20 mins by 1 mL of 5% spironolactone cream, then apply 1 mL of 2.5% AHK-Cu six hours later - after the minoxidil has reached full absorption at the ~4 hour mark, with some extra time allotted in between for any possible delayed absorption from the presence of the spironolactone cream. Then at night, I'll apply the minoxidil at T-7 hours before bed, followed by spironolactone, then another 1 mL of AHK-Cu six hours later, at T-1 hour before bed so it also has some time to dry. It puts a bit more strain on the schedule, but planning ahead like this for regularity and predictability makes that a feasible proposition.

Anything I'm missing here (besides the risk of the mid-day AHK-Cu application turning my scalp blue/green/yellow)?

 

[sizzlinghairs]

I was thinking about ordering some AHK, but I dont know what happened to finasteride fight er???

 

[TheLastHairbender]

Well, I think the math is right but it's not an altogether satisfying proposition:

Looking at the Pyo et al (2007) study on cultured dermal papilla cells ('the Korean study') they found positive results for treatment with .000000000001 to .00000001 moles of AHK-Cu (10^-12 to 10^-9 moles), but they resulted in cell death at concentrations of .00000001 and .0000001 (10^-8 and 10^-7 moles).

The amounts proposed for my formulation above deliver more moles than all of those treatment groups. The question is: what is the appropriate scaling up of quantity delivered to the cultured DPCs for application to the entire scalp? The treatment results in the study suggest there is a razor's edge point of optimality, above which you can actually induce the death of your dermal papilla cells, which is obviously the worst possible outcome.

I'm not sure why the negative treatment effects in the oft-cited study haven't gotten any debate among experimental users on these boards, but it's making me a bit uneasy about the at-home use of AHK-Cu. At this point I think I'm going to start with a much lower concentration, possibly even .25% AHK-Cu for a month or so to determine the safety of the project, and then scale up towards the 2.5% concentration I suggested before with careful monitoring.

I'm open to anyone else's thoughts or concerns about this, so please share your knowledge and experiences.


A few other facts about AHK-Cu for the interested party: storage should be at 2-8* C, in a sealed, dry, and dark container. (This equates to 35.6* - 46.4* F, so keep any unmixed powder in the fridge). I'll be making a solution of ethanol/PG/distilled water and adding the AHK-Cu within the next week or two and will be sure to post my notes on the process here for reference. Thanks guys.

 

[armandein]

Very interesting issue,TheLasthairbender, I am tunning on with your post. I am with you with the maths, There is great differences between the study and the amount of the doses with marketed lotions,
Good luck

 

[blaze]

Instead I'll bear the inconvenience for safety's sake and use 1 mL at 2.5% strength twice daily.

All this calculation is great. But Tricomin in their human studies that went to phase 2 in FDA trials used a 2.5% strength AHK-Cu topical for 6 months twice per day. No side effects were noted.

You went to all this trouble calculating all this and you came to the conclusion that is already available and established from human studies by the makers themselves, Tricomin.

 

[czvezda]

All this calculation is great. But Tricomin in their human studies that went to phase 2 in FDA trials used a 2.5% strength AHK-Cu topical for 6 months twice per day. No side effects were noted.

You went to all this trouble calculating all this and you came to the conclusion that is already available and established from human studies by the makers themselves, Tricomin.

Can you provide the link for that Tricomin study? I was not able to find it on http://clinicaltrials.gov

Thanks

 

[jonson]

What do you say about just add 2.5 gram of ahk into emu oil?
Could it work?

 

[czvezda]

What do you say about just add 2.5 gram of ahk into emu oil?
Could it work?

I don't think so. AHK-Cu dissolves well in water since it is polar, probably will not dissolve in lipids.

 

[Soldering]

Does anybody have a clue about an alternative source for AHK-CU now that r.axus.chem has appeared to have gone offline?

I'm in the UK and am only after 5g, just to try the stuff.

I was so interested in it, but then all this bother seems to have happened which took the one source I was certain of out of the running.

Now I'm scratching my head and try as I might I don't seem to be able to locate any alternative.

If anybody knows of a dependable different source I would be very grateful if you could let me know.

I'm very happy to be PM-ed on this subject too if someone thinks they can help. :bravo:

Cheers!

 

[TheLastHairbender]

All this calculation is great. But Tricomin in their human studies that went to phase 2 in FDA trials used a 2.5% strength AHK-Cu topical for 6 months twice per day. No side effects were noted.

You went to all this trouble calculating all this and you came to the conclusion that is already available and established from human studies by the makers themselves, Tricomin.

Yeah but the question is how much of the 2.5% solution did they use? 2.5% is not a quantity of active ingredient, it's the proportion of the total solution that is active ingredient. So the application of .25mL of 2.5% solution is a lot different than the application of 1 mL of 2.5% solution. Get it? Read my first post: (paraphrasing) 'If you come and say use a 2.5% solution without relating that to the quantity to be applied then it's clear you're talking out your ***."

It's like I'm saying: I want to get drunk tonight, how much should I drink. All you're telling me is what proof liquor I should drink, no mention of how much. If you came and said: "Hey you should drink 8 shots of 80 proof liquor or 16 shots of 40 proof grocery store liquor or just 3-4 shots of 190 proof everclear", then that makes sense. Your contribution was basically: "drink 80 proof because that's how jack daniels makes it". Thank you for adding nothing but contempt to the discussion.

And yes, I'd always rather do the stoichiometry myself than listen to what some board poster thinks he remembers was the quantity from some study back in 1997. I recommend everyone do the same. Unless multiplication is too tricky for you, then I guess your stuck taking "blaze's" word for it.

Edited: to add an apology to blaze for taking things personally. It's a reflection of my belief that the initial round of experimentation with AHK-Cu was not scientifically conducted and the advice handed out about solution strengths bordered on reckless, as I heard many people saying to use 2.5% or 5% concentrations without ever mentioning how much of a given strength solution to apply.

 

[blaze]

I understand what you said. But I think its just logical to use the amount needed to cover the treated area, however much that may be.

They found that 1.25% did nothing for hair loss in terms of growth or stabilization. So we can conclude from that that too small amount isnt effetive for human hair.

 

[jonson]

I use the ahk For between month or two and yesterday i noticed a little weird bald spot, could it be because the ahk? Maybe it is not good for me?

 

[TheLastHairbender]

I suppose it could be...there's obviously insufficient information to be able to make that determination based on your comments, but it is the kind of thing I'd monitor for very carefully. Maybe take some time off the AHK if you're concerned...the problem is just that it takes so long for any treatment or discontinuation effects to manifest themselves visibly in your hair's appearance, so I, and many others, find it difficult to attribute the expression of particular characteristics to any one element of a treatment plan.

It is important to note that at concentrations above the ideal therapeutic dose, the Pyo et al (2007) researchers found that AHK treatment resulted in the death of the cultured dermal papilla cells under study. Now I don't think AHK is necessarily bad for you, but it demonstrates that the dose-response relationship may not be monotonically or even weakly increasing: it may have more of a hump-shape. How to achieve the maximum value along that hump? That's what we're all trying to figure out.

Please provide more details of your treatment with AHK-Cu and more information about the resulting bald spot, maybe someone can offer some insight and adding your experience to our shared knowledge helps enrich our collective understanding. Thanks and best wishes for a quick resolution.