Injectable Glutathione Recipe

[Sampei]

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

yeah but who knows how long it last with carnitine in the mix... maybe it degrade and oxidize very quickly

 

[Mac81]

yeah but who knows how long it last with carnitine in the mix... maybe it degrade and oxidize very quickly

Yeah I know man... I'm curious where that guy earlier in the thread came up with the recipe, and the info about it supposedly being some sort of protectant of the glutathione.... All this s***'s is kind of just guessing when you have no way of actually testing it..

 

[nullandvoid]

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.

Best guess is I'm running with some pretty high systemic inflammation right now due to all the oils I bought last year having 10% EO in them (undisclosed by source until privately admitted). EO in things that don't even need it. So anything I inject, subcutaneously especially, just has a tendency to piss tissue off. But that's all it is.. a guess. Be glad when I finish them all out, and I can move on to a different source. Maybe it's that, maybe not. But for the Glutathione, it just has a bite for me I've had to try and accommodate. It's certainly better than Amino Asylum's which is like injecting acid. Carnitine can sometimes too, but it's a bit random. Modern Amino's version has been pretty good, though.

 

[Mac81]

Understanding the Requirements:

• Pharmaceutical Grade Glutathione: It is crucial to use pharmaceutical-grade glutathione, specifically intended for injection, and not a dietary supplement or industrial-grade product.
• Sterile Compounding Environment: Sterile compounding must be conducted in a dedicated, clean environment, such as a sterile compounding pharmacy, to ensure the final solution is free from microbial contamination.
• Sterile Water for Injection (SWFI): Only use sterile water specifically labeled for injection when preparing the solution.
• Buffering Agent: The pH of the glutathione solution must be adjusted to a physiologically acceptable range (usually around 7.4) to minimize irritation and ensure proper absorption.
• Sterilization: The final glutathione solution must be sterilized, typically through filtration with a 0.22-micron sterile filter, to ensure its sterility.

General Steps (Illustrative Purposes Only - Consult with a Professional):

1. Calculate the Required Glutathione Concentration: Determine the necessary glutathione dosage and volume of solution for the intended application.
2. Prepare the Buffering Agent: Choose a suitable buffer system, like phosphate buffer (pH 7.4), and prepare it using sterile water.
3. Dissolve the Glutathione: Add the calculated amount of pharmaceutical-grade glutathione powder to a portion of the sterile water and stir until completely dissolved.
4. Adjust the pH: Carefully adjust the pH of the glutathione solution to the desired level using a sterile acid (like HCl) or base (like NaOH).
5. Add Buffer and Dilute: Gradually add the buffering agent to the glutathione solution while monitoring the pH. Then, dilute the solution to the final volume with sterile water.
6. Sterilize the Solution: Filter the final glutathione solution through a 0.22-micron sterile filter into a sterile vial.
7. Quality Control: Inspect the solution for clarity, absence of particulate matter, and proper labeling before use.

 

[Mac81]

I found this guys.. This is basically exactly what we are doing by adding NAOh and dissolving glutathione... The only step we aren't going is adding the phosphate buffer, which it says can be done after you have dissolved the glutathione with the NAOH... This seems pretty simple actually

 

[Mac81]

So you can do what I did the other day, Make a wicked strong solution with pure NaOH, set the pH.. add the phosphate buffer.. double check the pH. Anybody have any input? All you need is water and the phosphate to make the buffer... It seems pretty damn simple, This is basically exactly what we were already doing, minus adding the buffer at the end...

 

[Sampei]

Understanding the Requirements:

• Pharmaceutical Grade Glutathione: It is crucial to use pharmaceutical-grade glutathione, specifically intended for injection, and not a dietary supplement or industrial-grade product.
• Sterile Compounding Environment: Sterile compounding must be conducted in a dedicated, clean environment, such as a sterile compounding pharmacy, to ensure the final solution is free from microbial contamination.
• Sterile Water for Injection (SWFI): Only use sterile water specifically labeled for injection when preparing the solution.
• Buffering Agent: The pH of the glutathione solution must be adjusted to a physiologically acceptable range (usually around 7.4) to minimize irritation and ensure proper absorption.
• Sterilization: The final glutathione solution must be sterilized, typically through filtration with a 0.22-micron sterile filter, to ensure its sterility.

General Steps (Illustrative Purposes Only - Consult with a Professional):

1. Calculate the Required Glutathione Concentration: Determine the necessary glutathione dosage and volume of solution for the intended application.
2. Prepare the Buffering Agent: Choose a suitable buffer system, like phosphate buffer (pH 7.4), and prepare it using sterile water.
3. Dissolve the Glutathione: Add the calculated amount of pharmaceutical-grade glutathione powder to a portion of the sterile water and stir until completely dissolved.
4. Adjust the pH: Carefully adjust the pH of the glutathione solution to the desired level using a sterile acid (like HCl) or base (like NaOH).
5. Add Buffer and Dilute: Gradually add the buffering agent to the glutathione solution while monitoring the pH. Then, dilute the solution to the final volume with sterile water.
6. Sterilize the Solution: Filter the final glutathione solution through a 0.22-micron sterile filter into a sterile vial.
7. Quality Control: Inspect the solution for clarity, absence of particulate matter, and proper labeling before use.

So I have PBS 1x solution how much of it do I need to use? That's the thing I'm not understanding much. You use the NaOH to raise the PH and that's ok but how much of the PBS Vs sterile water do I need to add to buffer the whole solution?

 

[Sampei]

So I have PBS 1x solution how much of it do I need to use? That's the thing I'm not understanding much. You use the NaOH to raise the PH and that's ok but how much of the PBS Vs sterile water do I need to add to buffer the whole solution?

Ok I found out the answer, you use PBS 1x solution to dissolve the glutathione using the NaOH to raise the PH to 7/7.4 max and just use PBS 1x to top up to the total volume you want. So no need to use any sterile water, just use the PBS 1x unless one use PBS 10x in that case you dilute it with sterile water.

 

[Mac81]

So I have PBS 1x solution how much of it do I need to use? That's the thing I'm not understanding much. You use the NaOH to raise the PH and that's ok but how much of the PBS Vs sterile water do I need to add to buffer the whole solution?

I knew you were going to ask me that, lol I'm working on it now, It can't be that hard to find, I'll get back with you with what I find out, as long as you think we're kind of on to something..

 

[Sampei]

I knew you were going to ask me that, lol I'm working on it now, It can't be that hard to find, I'll get back with you with what I find out, as long as you think we're kind of on to something..

Already found the answer, if pbs solution is 1x just use that like it's sterile water if it's 10x then you need to dilute it in sterile water so that you have a 1x solution.

Just easier to buy PBS 1x and dissolve the Gluthatione into it.

 

[Sampei]

## **600 mg/ml Reduced Glutathione in 1x PBS with 0.9% Benzyl Alcohol (30 ml, pH 7–7.4)**

**Ingredients:**
- Reduced Glutathione: **18.0 g**
- Benzyl Alcohol: **0.27 ml** (0.9% of 30 ml)
- 1x PBS: **18.93 ml** (see note)
- NaOH (1 N): as needed for pH

**Note:**
Glutathione powder displaces 0.6 ml/g (18 g = 10.8 ml displacement).
PBS volume = 30 ml − 10.8 ml (glutathione) − 0.27 ml (BA) = **18.93 ml**

---

### **Steps:**

1. **Add** 18.0 g glutathione powder to a beaker.
2. **Add** 0.27 ml benzyl alcohol.
3. **Add** ~18 ml 1x PBS (reserve the rest for final adjustment).
4. **Stir** to dissolve.
5. **Adjust pH** to 7.0–7.4 with NaOH (add dropwise).
6. **Transfer** to a 30 ml volumetric flask or cylinder.
7. **Top up** to 30 ml with remaining PBS.
8. **Mix well** and aliquot as needed.

---

**Storage:**
Refrigerate (2–8 °C), protect from light/air.

---

*Yields 30 ml of 600 mg/ml glutathione in 1x PBS, 0.9% BA, pH 7–7.4.*

Looks quite easy gonna try it soon as I get the glutathione powder!

 

[Mac81]

24 bucks on eBay for 500 ml...

 

[Mac81]

## **600 mg/ml Reduced Glutathione in 1x PBS with 0.9% Benzyl Alcohol (30 ml, pH 7–7.4)**

**Ingredients:**
- Reduced Glutathione: **18.0 g**
- Benzyl Alcohol: **0.27 ml** (0.9% of 30 ml)
- 1x PBS: **18.93 ml** (see note)
- NaOH (1 N): as needed for pH

**Note:**
Glutathione powder displaces 0.6 ml/g (18 g = 10.8 ml displacement).
PBS volume = 30 ml − 10.8 ml (glutathione) − 0.27 ml (BA) = **18.93 ml**

---

### **Steps:**

1. **Add** 18.0 g glutathione powder to a beaker.
2. **Add** 0.27 ml benzyl alcohol.
3. **Add** ~18 ml 1x PBS (reserve the rest for final adjustment).
4. **Stir** to dissolve.
5. **Adjust pH** to 7.0–7.4 with NaOH (add dropwise).
6. **Transfer** to a 30 ml volumetric flask or cylinder.
7. **Top up** to 30 ml with remaining PBS.
8. **Mix well** and aliquot as needed.

---

**Storage:**
Refrigerate (2–8 °C), protect from light/air.

---

*Yields 30 ml of 600 mg/ml glutathione in 1x PBS, 0.9% BA, pH 7–7.4.*

Looks quite easy gonna try it soon as I get the glutathione powder!

Hell yeah!

 

[Photon]

PBS is nothing magical, its just a combination of

Sodium Chloride
Sodium Phosphate Dibasic Anhydrous
Potassium Phosphate Monobasic
Water (majority of it)

You can get it at different concentrations (1x, 20x), etc, it just means the % of those compounds above are different. You can even get it in powdered form if that's easier for storage. 1X probably wont dissolve much glut but that's where naOH comes in.

 

[Sampei]

PBS is nothing magical, its just a combination of

Sodium Chloride
Sodium Phosphate Dibasic Anhydrous
Potassium Phosphate Monobasic
Water (majority of it)

You can get it at different concentrations (1x, 20x), etc, it just means the % of those compounds above are different. You can even get it in powdered form if that's easier for storage. 1X probably wont dissolve much glut but that's where naOH comes in.

Yeah it doesn't need to dissolve anything that's the buffer, the dissolution comes from the NaOH as you said. It's quite simple, use the PBS to keep it stable long-term and the NaOH to adjust PH and permit it to dissolve at least to 600mg/ml

 

[Duke309]

Yeah it doesn't need to dissolve anything that's the buffer, the dissolution comes from the NaOH as you said. It's quite simple, use the PBS to keep it stable long-term and the NaOH to adjust PH and permit it to dissolve at least to 600mg/ml

My question would be, what time frame are we talking about of refrigerated reconstituted glutathione at 600mg/ml would PBS be required to keep it stable? 1 month, 6 months, 1 year?

 

[Sampei]

My question would be, what time frame are we talking about of refrigerated reconstituted glutathione at 600mg/ml would PBS be required to keep it stable? 1 month, 6 months, 1 year?

I know that the buffered vials you buy of glutathione are stable 100% for 30 days, I'm trying to run a test where we try to se degradation at 60/90/120 days so we will an answer on at least 4 months

 

[Sampei]

Refined the recipes for added stability and anti oxidation long term.

Can be scaled..I'll brew 30ml and leave it in the fridge for 6 months and send it to Jano to test after to see if it has oxides or whatever

**Reduced Glutathione Solution (600 mg/ml, 30 ml) with Preservatives and Antioxidants**

**Ingredients:**
- Reduced Glutathione: 18.0 g
- Benzyl Alcohol: 0.27 ml (0.9%)
- Disodium EDTA: 6 mg (0.02%)
- Sodium Bisulfite: 30 mg (0.1%)
- 1x PBS: to final volume (see below)
- NaOH (1 N): as needed for pH adjustment

 

[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.

Hey I just kind of got the ball rolling, most of the credit is due to sampei, he took all the bits and pieces and threw them all together into a sensible recipe that's seems like it's going to be quite effective. Lol

 

[Mac81]

Refined the recipes for added stability and anti oxidation long term.

Can be scaled..I'll brew 30ml and leave it in the fridge for 6 months and send it to Jano to test after to see if it has oxides or whatever

**Reduced Glutathione Solution (600 mg/ml, 30 ml) with Preservatives and Antioxidants**

**Ingredients:**
- Reduced Glutathione: 18.0 g
- Benzyl Alcohol: 0.27 ml (0.9%)
- Disodium EDTA: 6 mg (0.02%)
- Sodium Bisulfite: 30 mg (0.1%)
- 1x PBS: to final volume (see below)
- NaOH (1 N): as needed for pH adjustment

You try that stuff yet man? I know I said I wasn't going to screw around with it for a while, but I've been getting bored, I actually have everything ordered from your recipe that I didn't have and it's getting delivered this week. So I'm about to try your recipe out, if you haven't yet! I'm planning on just diluting 500 ml of pbs with the edta and sodium bisulfite, that way I don't have to be trying to weigh out sub milligram measurements. Probably going to do it this Saturday....