Injectable Glutathione Recipe

[Mac81]

All the baking soda does is make it foam like hell, and turn Cloudy as fuck.. The pH remains the same, And it doesn't dissolve any more glutathione.... It's not turning it crystal clear and dissolving more and more glutathione like you are theorizing... You need to buy some supplies online, And do some experimentals and then it will all make sense to you. The pH is what determines how much it can hold in solution plain and simple, If you can't bring the pH up it's not going to take any more into solution., not to mention you do not want to inject something with the actual pH of glutathione, you will be walking funny for a week. I don't know how else to explain it,NAOH does a fantastic job of what I just said, you can dissolve so much of it It's f****** crazy,

 

[Mac81]

Not all buffers you stated should be injected, some are not buffers as well.


That depends on whats in the buffer, and how concentrated it is. A buffer of 7 is not going to remain at 7 long when you add something highly acidic or alkaline. We are talking about 700mg/ml+..not 50mg/ml+ or 200mg/ml+

Not all buffers you stated should be injected, some are not buffers as well.


That depends on whats in the buffer, and how concentrated it is. A buffer of 7 is not going to remain at 7 long when you add something highly acidic or alkaline. We are talking about 700mg/ml+..not 50mg/ml+ or 200mg/ml+

I have glutathione recipes for everyone of the ingredients I listed above, I can post them for you if you wish, They are very detailed and explain exactly how to use each one of them to make glutathione, They are just a lot more complicated than what we are doing, And there's a lot more steps involved, I just didn't post the whole process for each one, I just cut and pasted the ingredients.

 

[Sampei]

I have glutathione recipes for everyone of the ingredients I listed above, I can post them for you if you wish, They are very detailed and explain exactly how to use each one of them to make glutathione

Please do

 

[Mac81]

Please do

Will do

 

[Mac81]

To create a buffered glutathione (GSH) solution, you need to choose a buffer that maintains a stable pH in the desired range and is compatible with GSH and your intended application.
Here's a general approach, incorporating the provided search results:
1. Choose your buffer system:
Phosphate Buffers (PBS or Potassium Phosphate): Commonly used and generally compatible with GSH. Note that phosphate buffers can influence GSH oxidation and radical formation, so consider this if your application is redox-sensitive.
Tris-HCl Buffers: Another popular choice for biochemical applications.
MES Buffer: A buffer system specifically designed for GSH assays, often used at a slightly acidic pH.
2. Determine the desired pH:
Consider the pKa of GSH's thiol group (around 8.5 - 9).
Choose a pH that keeps GSH mostly in its reduced (thiol) form if that's important for your application.
If you're working in a system where GSH will be involved in redox reactions, consider the optimal pH for those reactions.
3. Prepare the buffer stock solution:
Follow standard protocols for preparing your chosen buffer.
Make sure to use high-purity water (e.g., ultrapure water).
Adjust the pH of the buffer to the desired value using HCl or NaOH, as needed.
4. Dissolve GSH in the buffer:
Weigh out the required amount of reduced GSH (GSH).
Dissolve the GSH powder directly in the prepared buffer solution.
5. Consider adding stabilizing agents (optional):
EDTA: May help to chelate metal ions that can catalyze GSH oxidation.
Sulfosalicylic acid (SSA): Can help to deproteinize and act as an antioxidant, but it's important to note it may not be suitable for all applications.
Example using a phosphate buffer:
To prepare a 10 mM GSH solution in 100 mM potassium phosphate buffer, pH 7.0:
Prepare a 100 mM potassium phosphate buffer, pH 7.0.
Weigh out the appropriate amount of GSH (e.g., using the molecular weight of 307.3 g/mol for reduced GSH).
Dissolve the GSH in the phosphate buffer.
Adjust the pH, if necessary, to 7.0.
The final solution would be 10 mM GSH in 100 mM potassium phosphate buffer, pH 7.0.
Important considerations:
GSH oxidation: GSH is susceptible to oxidation, so minimize exposure to air.
Storage: Freshly prepared GSH solutions are generally preferred. If you need to store the solution, freeze in aliquots at -20°C or lower.
Compatibility: Ensure the buffer and any added agents are compatible with your specific assay or experiment.
Remember to consult the specific protocols and guidelines for your chosen buffer system and GSH assay or application.

 

[Mac81]

Tris-HCl as a Buffer:

• Tris-HCl is a widely used buffer in biochemistry and molecular biology.
• It helps maintain a stable pH, which is crucial for the stability and activity of biomolecules like glutathione.
• Tris buffers are effective in the pH range of 7.0 to 9.0, making them suitable for many biological applications.
• Using Tris-HCl prevents "overshooting" the desired pH during buffer preparation and avoids the need for strong acids or bases, according to Hopax Fine Chemicals.

2. Preparing Glutathione Solutions with Tris-HCl:

• Dissolving Glutathione: Reduced glutathione (GSH) is soluble in water and can be directly dissolved in Tris-HCl buffer at the desired concentration.
• pH Adjustment: If needed, adjust the pH of the Tris-HCl buffer to the desired value (often around pH 8.0 for applications like GST-fusion protein elution) before adding the glutathione.
• Elution Buffer Example: For example, a common elution buffer used in GST-tagged protein purification contains 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 0.1 mM EDTA, and 10 mM reduced glutathione.

3. Considerations:

• Glutathione Stability: While glutathione is more stable in water, solutions should be prepared and used fresh for optimal activity, especially if you're working with reduced glutathione (GSH), according to Bio-Rad.
• Storage: If storage is necessary, glutathione solutions are more stable at lower temperatures (e.g., -20°C).
• Specific Application: Always check the specific protocol or application to determine the appropriate Tris-HCl buffer concentration and pH for your needs.

 

[Mac81]

I'll post the other stuff I have later, I got to get back to work

 

[Mac81]

While baking soda (sodium bicarbonate) can act as a buffer in various situations, it's not the ideal choice for buffering glutathione solutions, particularly in biological contexts, due to potential side effects and limitations.
Here's why:
1. Baking Soda and pH Adjustment:

• Baking soda is a base and can increase the pH of a solution.
• Glutathione solutions, like those used in research or supplements, may need a specific pH range for optimal stability and activity.
• Using baking soda could lead to a pH that's too high (alkaline) for the desired application of glutathione.

2. Buffers and Maintaining pH Stability:

• Buffers are essential for maintaining a stable pH, resisting changes when acids or bases are added.
• Good buffers for biochemical reactions typically have a pKa (acid dissociation constant) within the desired pH range, are water-soluble, and don't interfere with biological processes.
• Baking soda's buffering range may not be suitable for all situations involving glutathione, and its use could lead to unwanted side effects.

3. Considerations for Glutathione Buffering:

• Specific Buffer Systems: In biological systems, the bicarbonate buffer system is crucial for maintaining blood pH, but other buffer systems, like phosphate buffers or organic buffers like HEPES, are often used in research and pharmaceutical applications, each with specific properties and advantages.
• Potential Side Effects: Sodium bicarbonate can lead to various side effects, including gastrointestinal issues, electrolyte imbalances, and acid-base disturbances, especially with chronic or high-dose use.
• Interactions with Glutathione: There's no specific evidence suggesting direct chemical interactions between baking soda and glutathione. However, changes in pH caused by baking soda could indirectly impact glutathione's stability or activity.

In summary:

• While baking soda can buffer pH in certain situations, it's not the ideal choice for buffering glutathione, particularly in biological applications.
• Consider using established buffer systems suitable for the specific application of the glutathione solution, and be mindful of potential side effects.
• Consult with a healthcare professional or relevant expert for specific advice regarding buffering glutathione for particular applications.

 

[Mac81]

As photon already said phosphate buffers are generally preferred in pharmaceutical applications and work the best, but you still have to adjust the pH with NaOH after you buffer it... So if you wanted to get all technical the very first one I posted Is as accurate as you can get with what the pharmaceutical industry does, And it is the most accurate recipe I posted, And if you notice it says to adjust the pH as needed, so as you increase the concentration you're going to need to continue to adjust the pH, It's a simple give and take, the more glutathione you add, The more NAOH you need, you can buffer it with all kinds of stuff according to your intended application and whatever you feel works best, but you can't get around the pH thing when you continue to jack up the concentration, the buffer is limited in how much It can manipulate the pH, you can only put so much buffer in before you start overriding your actual concentration, The pH drives the f****** car, the buffer is just a passenger, or the seat belts, however you want to look at it

 

[Photon]

I have glutathione recipes for everyone of the ingredients I listed above,

Will not comment on your recipe, but i hope you at least tested it first and not pulled it out from ChatGpt. Some of these buffers..are not for injections.

 

[Mac81]

Will not comment on your recipe, but i hope you at least tested it first and not pulled it out from ChatGpt. Some of these buffers..are not for injections.

No I would definitely go with the phosphate one, as that is what the pharma industry uses, I got that from sigma Aldrich,And if you notice there's one reoccurring theme, It doesn't matter what you buffer it with It's still doesn't store long-term worth a f*** unless you freeze it, which is why it comes in a vial and you have to reconstitute it to use it.

 

[Mac81]

We are basically chasing a pipe dream, you can buffer with this, you can buffer it with that, This buffer will do this, This buffer will do that, none of them do the same thing, none of them prevent oxidization, which is what we're ultimately chasing here with this 90 f****** replies over baking soda, The ultimate buffer with the most health benefits, that last longest, which is simply something that doesn't exist with pre-made glutathione solutions, you make them and you use them, or you freeze them, there is no miracle recipe, And if you don't want a weak ass concentration, you must change the pH with a strong base... It's just the writing on the wall

 

[Mac81]

I appreciate everything everybody's contributed, but I for one have no intention of changing anything, If it's not broke don't fix it, plenty of people here already figured out what works and what works well, And it's simple just don't make too much of it and plan on storing long term Make what you need.... I'm bowing out once and for all, lol thanks for everybody's input and good luck... If anything I learned why it comes in little glass vials with no liquid, It's the only f****** way to make it last any amount of time

 

[Mac81]

And there's no way in hell I'm Jano testing My homemade glutathione 60 days in to see how much it oxidized, lmao.. It's quite obvious the pharma industry sells it unconstituted for a reason... I think we're trying to reinvent the wheel here.

 

[nullandvoid]

I just like the simplicity of bacteriostatic NaCl with nothing else (with the Glutaone vials, which have some unknown amount of sodium bicarbonate as a buffer). 11ml of bacteriostatic NaCl creates roughly a 100mg/ml solution which is good enough for my uses (the huge puck itself adds about 1ml to the final volume). It still has a very mild site reaction, but it's tolerable. Bacteriostatic water requires I fill nearly the entire 20ml vial, requiring well over 1ml for 100mg, and it still produces a worse site reaction than the NaCl. But as mentioned, I've since learned about the oxidation and how fragile, so I've been doing the aliquot method and freezing syringes. NaCl is not an ideal choice for the sake of oxidation, but by freezing them immediately, I can get close to the best of all worlds... reduced volume, lowest site reaction, and preservation of the Glutathione.

 

[Mac81]

I don't know what you guys think... But I do think the next time I make it I'm going to put it in syringes and freeze them individually... That seems about the smartest way to store this stuff long-term and guarantee that it doesn't lose any of its effectiveness?

 

[Mac81]

I think the recipe with the L-Carnitine has some uses though.. sampei had mentioned it, A lot of people are trying to take both compounds anyways and a 600/ 200 solution is admirable.. And supposedly whoever came up with that claims that L-Carnitine buffers it and makes it last much longer.... It's damn simple you don't even need NAOH... You just add all three things in a beaker and stir it up and it comes out crystal clear you can't get much simpler than that It was the first one I ever made.. I think I would rather do that than try to figure out the baking soda secret... I see no need to inject baking soda, L-Carnitine and glutathione though? Seems like a pretty good combo..It's a solid recipe and it plain works.. still got to freeze it though.. I don't see any way around that, nothing completely protects it regardless how you make it, It's a perishable item. And I'm willing to bet you could tweak that recipe any way you wanted with pure NAOH . And make it any ratio you wanted

 

[Sampei]

I just like the simplicity of bacteriostatic NaCl with nothing else (with the Glutaone vials, which have some unknown amount of sodium bicarbonate as a buffer). 11ml of bacteriostatic NaCl creates roughly a 100mg/ml solution which is good enough for my uses (the huge puck itself adds about 1ml to the final volume). It still has a very mild site reaction, but it's tolerable. Bacteriostatic water requires I fill nearly the entire 20ml vial, requiring well over 1ml for 100mg, and it still produces a worse site reaction than the NaCl. But as mentioned, I've since learned about the oxidation and how fragile, so I've been doing the aliquot method and freezing syringes. NaCl is not an ideal choice for the sake of oxidation, but by freezing them immediately, I can get close to the best of all worlds... reduced volume, lowest site reaction, and preservation of the Glutathione.

that's just bullshit, I reconstitute 2400mg vials of glutathione with 3ml BAC and it comes out as 4ml of 600mg/ml solution.

I inject 1ml twice a week.

your statement of: well over 1ml for 100mg if you use BAC it's utter nonsense lol

You talk about freezing but have you ever lab tested if you can freeze reconstituted glutathione without harming the substance itself? no you have no idea, so that's another assumption you are making that is not proven at all.

Glutathione can stay in solution 30 days without any significant degradation so there is that, more I have no clue but I'll try to find out with a nice degradation test

 

[Photon]

that's just bullshit, I reconstitute 2400mg vials of glutathione with 3ml BAC and it comes out as 4ml of 600mg/ml solution.

I inject 1ml twice a week.

your statement of: well over 1ml for 100mg if you use BAC it's utter nonsense lol

You talk about freezing but have you ever lab tested if you can freeze reconstituted glutathione without harming the substance itself? no you have no idea, so that's another assumption you are making that is not proven at all.

Glutathione can stay in solution 30 days without any significant degradation so there is that, more I have no clue but I'll try to find out with a nice degradation test

We are able to reconstitute at such high mg/ml because it is already buffered.

Whereas when brewing, you need to keep it within a certain pH range just to dissolve.

I was planning to brew something like 100ml but then I'm not entirely sure if it will degrade. A good test is a glutathione blood test, which can be taken monthly. ($60 in the US). However at that point, it might be better to just buy it rather than brew.

I just like the simplicity of bacteriostatic NaCl with nothing else (with the Glutaone vials, which have some unknown amount of sodium bicarbonate as a buffer).

I'm not sure what are you doing with Glutaone, I am using the exact same product. I reconstitute it with hospira bac water, 700+ mg/ml and i pin it subq 0.15ml everyday using a 33g 4mm needle. Why are you struggling with such low concentrations, I have no idea.

Even doing 1ml at 700+ mg/ml subq has no pip for me.

 

[Sampei]

We are able to reconstitute at such high mg/ml because it is already buffered.

Whereas when brewing, you need to keep it within a certain pH range just to dissolve.

I was planning to brew something like 100ml but then I'm not entirely sure if it will degrade. A good test is a glutathione blood test, which can be taken monthly. ($60 in the US). However at that point, it might be better to just buy it rather than brew.



I'm not sure what are you doing with Glutaone, I am using the exact same product. I reconstitute it with hospira bac water, 700+ mg/ml and i pin it subq 0.15ml everyday using a 33g 4mm needle. Why are you struggling with such low concentrations, I have no idea.

Even doing 1ml at 700+ mg/ml subq has no pip for me.

I was referring to his statement of using premade glutathione lyophilized vials and not being able as you clearly stated to have 600-700mg/ml solution. it's utter nonsense we both just stated it

I well know that when we do our own glutathione the solution can be more or less concentrated depending on the buffer we use, I have tested buffered glutathione and at 30 days there was 0 degradation. Not sure past 30 days tho.

Not sure we can achieve the same buffer tho... I'm conflicted on what kind of buffer use, maybe a mix of NaOH and sodium bicarbonate?

If I can''t achieve 600mg/ml it becomes kinda useless to brew it myself, as I don't like to pin everyday so those 600mg/ml x2 a week is what I wanna achieve.