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Methylene Blue Dosing and Products


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#241 MrHappy

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Posted 18 October 2011 - 11:29 AM

Looks alright, from the websites...
24.4mg/mL is your starting point, therefore 1 drop (50mcL) is approx 1.2mg. If you put that into 1L of water, then 50mL of the final solution = 60mcg.
Or better yet, use a syringe and do it accurately.

#242 manic_racetam

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Posted 20 October 2011 - 06:43 PM

Sorry, I should have clarified that I don't mean in regard to the dosage. Does taking the undiluted powder put you at a higher risk for mutagenic effects? Or is that only a concern for contact with the powder to skin?

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#243 MrHappy

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Posted 20 October 2011 - 08:48 PM

The raw powder is somewhat hazardous. Lab precautions apply. Gloves and mask.

Not sure why you went for powder, when 4mL of 2.44% would get you to 100mg. Be careful.




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#244 amark

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Posted 20 October 2011 - 11:01 PM

Well here is a massive amount of information on handling large amounts of the substance. So far I have not had any potentia gastric side effects such as diarrhea vomiting or nausea. I certainly will make every effort to handle it carefully. After my supply is gone I will consider using a non powder form.

SPI Supplies Division
Structure Probe, Inc.

P.O. Box 656 West Chester, PA 19381-0656 USA
Phone: 1-(610)-436-5400 Fax: 1-(610)-436-5755
E-mail: spi3spi@2spi.com

WWW: http://www.2spi.com
Manufacturer's CAGE: 1P573

Material Safety Data Sheet

SPI #02573-AB/02573-MB SPI-ChemTM Methylene Blue
Posted Image


<Link>

[edited out the wall of text, it was a bit too long]

Edited by chrono, 15 November 2011 - 06:23 AM.


#245 chrono

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Posted 25 October 2011 - 09:08 AM

I just finished re-rendering the big MB threads; I'm bumping them all so everyone will know where they are.

Methylene Blue Experiences
Methylene Blue Dosing and Products
Methylene Blue Research
Methylene Blue and MAO Inhibition

These topics are interrelated, but we should try our best to keep discussions as topically segregated as realistically possible, to make the information easier to find in the future.
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#246 chrono

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Posted 15 November 2011 - 07:12 AM

I've been trying to think of a way to get this stuff into a capsule for a while. Partly for my own convenience, but also because I'd like my parents to take it, and even getting them to remember to take their fish oil is a war of attrition.

As we all know, normal gelatin capsules dissolve fairly quickly in water, so this is a difficult problem. My best general idea has been to drop it into a capsule filled with powder, hopefully preventing contact between water and the inside of the capsule. Success depends upon how well the powder absorbs the liquid.

I just tried it with ALCAR, which is fairly aggressively hygroscopic. It instantly shrinks away from the sides of the capsule into a bright blue clump. I place the drop with the capsule about 3/4 loosely full, then pack more on top before sealing it. After about an hour, the capsule has lost no integrity, and the blue band isn't even sticking to the sides. However, I'm not sure about this in the longer-term. In the summer humidity, ALCAR capsules become goopy messes. Will this occur with humidity inside the capsule, or must it be "pulled through" by ALCAR's hygroscopy (storing in a bottle with a silica gel pack might do this)?

I think I'll make some capsules with different powders inside, and see how they fare in storage. I have some supplements which come with veggie caps, which I think might be more resistant to water, though I'm not confident. Does anyone else have any clever ideas on this subject? Ascorbic acid is also hygroscopic, and might create the leuco MB form as a bonus.

EDIT: The first capsule I made with ALCAR was a fluke. The others began sticking to the sides instantly, and are slowly making the capsule softer. Sugar is the same story. Flour is non-absorbent, but a drop beads very nicely, usually contacting the side in only a very small area once compressed (though it eats through almost instantly). The only success I've had so far is by filling a cap loosely with flour, then creating a cavity by inserting a thin cylinder which leaves a good layer all around the hole, and then compressing more flour once I've inserted the drop. There's no contact with the sides. This is obviously a fussy method, though a capsule machine speeds up the process a bit.

EDIT 2: Actually, the ALCAR and sugar capsules lasted longer than I thought they would. They're both still fine after 12 hours, though some are starting to become more flimsy where there's obviously more MB in contact with the cap (darker stain). Sugar is actually working quite a bit better than ALCAR. I wouldn't consider this for long-term storage, but it would probably work just fine if you wanted to make up a dose to take while you're out of the house later in the day, for instance.


A significant improvement to this method would be to drop using a micropipette. This would involve less water, with the added benefit of probably being more accurate than the clumsy glass droppers we're all using. I found this 5uL mini pipette for $30 with a quick google search, and it looks like the company will sell to individuals. Not as good as the fancy 0.5uL micropipettes, but 10x smaller than a normal water drop, and it also doesn't cost $250. Could someone who knows more about these explain the function of the slide-on plastic tip? This pipette fits 200uL and 20uL tips, but supposedly delivers 5uL drops. Is the drop size independent of the tip? Is it even necessary?

Edited by chrono, 15 November 2011 - 10:52 PM.


#247 medievil

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Posted 15 November 2011 - 10:31 PM

The raw powder is somewhat hazardous. Lab precautions apply. Gloves and mask.

Not sure why you went for powder, when 4mL of 2.44% would get you to 100mg. Be careful.

I got 100mg for 2,5 euro's from a belguim pharmacy, i beleive someone here mentioned its solled in belguim in pharmacy's, cheers for the tip haha, i tried fishstores first but they were only selling rubbish with other shit.

#248 Metrodorus

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Posted 20 December 2011 - 01:37 PM

Having read through this forum, and ploughed through quite a number of articles - nice summary here:
http://www.dose-resp...nzalez Lima.pdf
I am a bit perplexed at the dosages that are being mentioned by some contributors to this forum:
The first is, that 60 mcg dosage - is that supposed to be 60 micrograms per kg of body mass? That would make a bit more sense.

The dosage of 1 mL for a standard human of 70kg of a 1:9 dilution of a standard 1% solution seems more like a reasonable dose, to
obtain the levels of MB in blood plasma seen in the cited studies, when accounting for differences in metabolism between humans
and small mammals...

So....can someone give a reference for the 60 mcg dosage,if it isn't calibrated per kg of body mass? It seems lower by an order of magnitude than the dosages that are having an effect in the animal studies.

#249 Logic

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Posted 20 December 2011 - 02:14 PM

Hey everyone :)

Im in the process of moving and have just recieved my mb.
Theres a lot to do and unfortunatly just dont have time to re-read this informative thread , so your assistance would be greatly apreciated.

The concentration is 12mg/ml ( Thats not right is it?)

As i understand it there are different dosages from life extension to anti-malerial etc.

So my quick question is how many drops in how much water for the life extension and mental acuity effect considering I take multivits including C and resveratrol.

thx guys (I hope! :) )

Ill intro myself better after the move.

Edited by Logic, 20 December 2011 - 02:41 PM.


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#250 niner

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Posted 20 December 2011 - 03:06 PM

So....can someone give a reference for the 60 mcg dosage,if it isn't calibrated per kg of body mass? It seems lower by an order of magnitude than the dosages that are having an effect in the animal studies.


Thanks for that great ppt, Metrodorus. It's a very good presentation. I agree that the 60ug dose seems too small. It's used here as the total dose, not per kg. As far as I can tell, this dose was essentially pulled out of a cocked hat. Some of us have been using larger doses, but only a couple people reporting here are dosing at a level that seems sufficient to hit the blood levels talked about in the literature. One problem is that those levels can get into the MAOI range; the ic50 for mao-a is 164 nM, iirc. Another problem is that most of us don't have what I'd consider a safe product for chronic use at high dosages. I don't have a problem with taking microgram doses of "fish medicine", but I don't know enough about its purity to be comfortable with milligram dosing.

I have a strong suspicion that there's a lot of placebo effect here...

Edited by niner, 20 December 2011 - 03:07 PM.


#251 Metrodorus

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Posted 20 December 2011 - 03:24 PM

Thanks.
I am collecting a selection of literature on the subject in a little blog post I put together:
http://whatifwhen.bl...eep-eye-on.html

I suspect the 60 microgram dose was at first intended as a dosage per kg body mass - otherwise there is too much variability.
All dosages should really refer to body mass.....otherwise there is no way of actually making comparisons between experimental subjects.

I am interested in MB because of its effect on mitochondrial efficiency.

#252 niner

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Posted 21 December 2011 - 01:50 AM

Metrodorus, you should check out this guy:

http://www.ncbi.nlm....term="Atamna H"[Author]

#253 MrHappy

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Posted 21 December 2011 - 05:16 AM

Niner, I would heavily suggest non-placebo effects were observed at 60mcg, which were considerably more pronounced at 1mg.

However, this seems to vary, greatly, between subjects.





#254 MrHappy

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Posted 21 December 2011 - 05:27 AM

Logic - 12mg/mL is correct.
Get a 1mL syringe and measure it out / dilute it in water far more easily.

To make an intermediate solution,
1:10 = 1.2mg/mL
1:200 = 60mcg/mL

Then take 1mL of the intermediate solution in water per dose.



#255 Metrodorus

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Posted 21 December 2011 - 11:46 AM

1 microgram is one millionth of a gram.
Required Dilutions:

1.2g per 100mL original 1.2% solution w/v
irrespective of original volume,we dilute as follows:
FIRST DILUTION
one volume of the original 1.2% solution to nine volumes of water
This shifts the decimal one place so we get
0.12g per 100mL

SECOND DILUTION
Take one unit volume of this new solution,and add to it 9 volumes of water.
This shifts the decimal a further place ,
0.012g per 100mL
THIRD DILUTION
Take one unit volume of this second dilution,and add to it 9 unit volumes of water.
This shifts the decimal yet a further place :
0.0012g per 100mL

1mL of this third dilution solution will contain 0.000 012g, or 12 mcg of solute.
5mL of this solution will contain 60mcg of solute.

Edited by Metrodorus, 21 December 2011 - 11:48 AM.

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#256 Metrodorus

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Posted 21 December 2011 - 01:19 PM

http://www.philiphar...blue-1-aqueous/

This is a standard 1% solution w/v

#257 niner

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Posted 21 December 2011 - 09:56 PM

Niner, I would heavily suggest non-placebo effects were observed at 60mcg, which were considerably more pronounced at 1mg.

However, this seems to vary, greatly, between subjects.


But as far as I know, no one has ever done a placebo controlled experiment. We all see the brilliant blue color, drink it, and feel all sorts of effects. Or not. Some of us never felt anything, or eventually came to the conclusion that we weren't feeling anything that was real. Between that and the fact that we aren't taking enough to generate the levels that all the papers say you need (~100 nM or more), I just don't see what else it could be.

#258 Metrodorus

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Posted 21 December 2011 - 11:21 PM

What is more, Niner, looking over the various forum posts on MB, many people here seem to be thinking they are taking mcg dosages, but their dilution methodology is dodgy, to say the least - 60mcg will not colour the water.

No-where in the literature that I have seen is there anything approaching an oral dosage of 60 mcg. Yes, some in vitro studies used nanogram concentrations - but with a 60 mcg dosage orally, you would not reach nanogram plasma concentrations.

Most of orally administered MB will simply bind immediately to haemoglobin, or to the intestinal epithelium, and some 10 - 20% binds to organic matter in the intestines, and is excreted in the stool. Only a tiny fraction of that 60mcg oral dose - picograms probably - will cross the blood brain barrier. I just don't see how a 60 microgram dose can do anything therapeutic - although MB does accumulate in mitochondria, so one *might* see an effect gradually build up over many days.

1000 mcg gives the water the colour of blue sapphire gin, and after absorption, nanograms are present in the blood plasma - around 100 or so, give or take - even with a 100% margin of error, this is still a low blood plasma level of MB, and is possibly more or less what we might be aiming for, to achieve the positive hormetic effect seen at low dosages.

Personally, at a 1mg dosage for a 70kg human, if you took a memory test, or a resistance training test, there might be a statistically significant result, and a weight lifter might notice a difference, but there might be no significant result at all.......

BTW, I don't see the point of reducing the MB with an acid prior to ingestion - just wash it down with an alpha lipoic acid cap. The stomach acid will reduce it anyway, so even that is probably a waste of time. Most of the MB in vivo apparently cycles back and forth between the reduced and oxidised state anyway, and most of it is reduced naturally. Around 70% of the MB excreted in the urine is in reduced when administered at much higher dosages than proposed here. At a low dose of 1000 - 2000mcg (depending on your body mass), maybe almost all of it is reduced, and this would require testing.

Apologies for the lack of footnotes.
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#259 Metrodorus

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Posted 21 December 2011 - 11:57 PM

Posted Image
What dilutions of MB look like starting with a 1g/100mL solution. (1 000 000mcg) (not shown)
one volume of this in nine volumes of water: 0.1g/100mL (100 000mcg) (not shown)
one volume of this in nine volumes of water 0.01g/100mL (10 000mcg)
one volume of this in nine volumes of water 0.001g/100mL (1000mcg)
one volume of this in nine volumes of water 0.0001g/100mL (100mcg)
one volume of this in nine volumes of water 0.00001g/100mL (10mcg)


I did not do a further dilution to 1 mcg, as at 10mcg the solution was already indistinguishable from tapwater.

#260 Metrodorus

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Posted 22 December 2011 - 11:28 AM

Posted Image

#261 sapentia

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Posted 28 December 2011 - 02:58 PM

I've been trying to think of a way to get this stuff into a capsule for a while. Partly for my own convenience, but also because I'd like my parents to take it, and even getting them to remember to take their fish oil is a war of attrition.


I just started my parents on it yesterday though not with capsules. I mixed a solution for them, put it in a medicine dropper bottle, and all they have to do is add 1 dropper full into their drink to get roughly 60 mcg. I will post back if they have any significant effects.


But as far as I know, no one has ever done a placebo controlled experiment. We all see the brilliant blue color, drink it, and feel all sorts of effects. Or not. Some of us never felt anything, or eventually came to the conclusion that we weren't feeling anything that was real. Between that and the fact that we aren't taking enough to generate the levels that all the papers say you need (~100 nM or more), I just don't see what else it could be.


I started taking MB yesterday, two doses at 60 mcg; 12 hours later at 6:00am I woke up with the worst headache I've had in years. There was no placebo about it and I am sure it was from the MB. The other effect was initially after dosing was akin to a stimulant feeling. The complexities with why MB would cause a headache at such a low dose are in my case compounded by the twenty plus other supplements I take. I can only imagine what would happen if I took a milligram dose like some others on this forum. Nevertheless, I will continue to take it and closely monitor any perceived effects; I have done this for years with other supplements so I am perhaps not quite as susceptible to a placebo effect. As always, we shall see. As noted above, I will also report on any changes with my parents; my mother in particular suffers from considerable fatigue and muscular weakness stemming from a minor form of Chiari Malformation.

#262 niner

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Posted 29 December 2011 - 04:20 AM

There was no placebo about it and I am sure it was from the MB.


Why are you sure? Maybe you just had a headache. I don't doubt it was a real headache that hurt like hell, but the only thing tying it to MB is proximity in time. If on multiple occasions it were to happen reliably after using MB, and only after using MB, then you'd have better evidence.

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#263 niner

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Posted 29 December 2011 - 05:19 AM

BTW, I don't see the point of reducing the MB with an acid prior to ingestion - just wash it down with an alpha lipoic acid cap. The stomach acid will reduce it anyway, so even that is probably a waste of time. Most of the MB in vivo apparently cycles back and forth between the reduced and oxidised state anyway, and most of it is reduced naturally. Around 70% of the MB excreted in the urine is in reduced when administered at much higher dosages than proposed here. At a low dose of 1000 - 2000mcg (depending on your body mass), maybe almost all of it is reduced, and this would require testing.


If I remember correctly, it's been shown that more MB ends up in the brain if it's dosed in the reduced form. At any rate, the pharmacokinetics are certainly going to be different between the oxidized and leuco forms. A lot can get reduced in the body, as you point out, though I suspect the time course of this is slow, since reduction by ascorbate in solution is slow. Stomach acid won't reduce MB; it's not a reducing agent.

Posted Image


This looks to me to be off by a factor of ten, based on what I've seen. Did you start with crystalline MB and dissolve it yourself, or did you start with a 1% by weight solution? Is it possible that some of this is in the leuco (reduced) form? When I've done dilutions, mine were darker in color, such that my dilutions looked like yours of the next higher concentration. I started with a 2.303% solution from Kordon. I know that some people here have messed up their dilutions, but I'm a chemist, so I have this sort of thing pretty well at hand.

What size are these glasses? They look like smallish tumblers, maybe 12oz? Or they could be tall shot glasses... If they are small, then maybe that's making them look lighter.

#264 smithx

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Posted 29 December 2011 - 11:34 AM

I've been trying to think of a way to get this stuff into a capsule for a while.


Here's one method which may work:

- Figure out the weight of the amount of some inert substance like corn starch which will fit into a capsule. Its probably best to fill 10 or more capsules, then empty them into a container and weigh the total, then divide by 10 so that you get a good average.
- Measure 1000 times that amount and put it into a container.
- Add to the container the amount of MB solution required for 1000 doses
- Mix well
- Allow the powder to dry
- Mix it again so that all the MB is very evenly distributed
- Fill capsules with the dry powder as needed
- Save the rest of the powder for later
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#265 sapentia

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Posted 29 December 2011 - 03:12 PM

There was no placebo about it and I am sure it was from the MB.


Why are you sure? Maybe you just had a headache. I don't doubt it was a real headache that hurt like hell, but the only thing tying it to MB is proximity in time. If on multiple occasions it were to happen reliably after using MB, and only after using MB, then you'd have better evidence.


My surety lies in the fact that I almost never get any form of headache, and I haven't had a severe headache in years. Granted, it is merely a probability and not a conclusive point of fact. That being said after again dosing MB yesterday 3 times at roughly 60 mcg I had no headache. I speculate that perhaps MB's antimicrobial and anti-fungal properties could have flushed out some impurities from my GI tract and other systems with toxins triggering the aforementioned headache.

#266 Metrodorus

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Posted 31 December 2011 - 04:02 PM

"This looks to me to be off by a factor of ten, based on what I've seen. Did you start with crystalline MB and dissolve it yourself, or did you start with a 1% by weight solution?

1% w/v solution from a reputable laboratory supply company. It was a fresh batch.
Dilutions were made as described.

What size are these glasses? They look like smallish tumblers, maybe 12oz? Or they could be tall shot glasses... If they are small, then maybe that's making them look lighter.
35mm diameter glasses.

#267 sapentia

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Posted 11 January 2012 - 06:03 PM

An update:

After ten days or so of supplementing with MB (60mcg initial to 300 mcg final) I ceased taking it due to it having a definite cognitive suppression effect. This was dissappointing as cognitive improvement was one of the effects I was hoping for as I had seen others report improved memory as a perceived benefit. Without knowing any specific mechanisms I speculate my personal results may have stemmed from an effect on 5-alpha reductase and the conversion of testosterone to DHT. I say this because other compounds which affect this pathway, soy and finasteride, have had a similar suppresion effect on my cognitive functioning. That combined with a loose knowledge that MB has a negative effect on sexual function is the basis for my speculation. Regardless as to the mechanism for my individual response to MB a result is a result and anything which adversely affects my mental fitness is unacceptable for supplementation.

On the positive side, I did notice what I would describe as a measure of increased vitality throughout the rest of my body. This particular effect is more subjective in my opinion, nevertheless, all things considered it falls within likely benefits.

Also, both of my parents are still on MB at an approximate daily dose of 350 mcg in two doses at 175 mcg each. Neither of them has indicated any negative side effect and both seem to have a measure of increased stamina and vigor though this is highly subjective. However, as noted in an earlier post my mother suffers from a mild level of Chiari Malformation Syndrome and has very significant fatigue and strength issues allowing for an easier determination of supplementation effectiveness. It is acknowledged that any perceived improvement is subjective and not valid for any scientific basis. My plan is to increase my parents dosage over a couple months until they are at roughly 1,000 mcg - 1,500 mcg per day as that seems to be the agreed upon dose for optimal results.

#268 Logic

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Posted 13 January 2012 - 05:40 PM

Thx for the replies guys.

I mixed 1 drop into 1.2 Liters of water and have been taking just under 100 ml to get around 1mg per dose, twice a day.

I have seen a definate improvement in memory and cognitave function.
My folks think its the bomb and its refreshing to see em actually awake when I visit. :)

#269 hippocampus

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Posted 16 January 2012 - 07:33 PM

I have 0,3% solution. How much drops do I need to put in 0,25 L of water to get 60 mcg of MB in 1 drop?

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#270 Metrodorus

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Posted 16 January 2012 - 07:38 PM

What is this obsession with drops? It is a really inaccurate way of dosing.
60mcg per 100mL would be more sensible to aim for, when you are at such miniscule concentrations.
Why would you want to take 60mcg anyway?
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