Typical protocols people are going through (in a step by step basis) for use of grey market peptides

I’m working on learning more about peptides and the protocols associated with their use. Gemini wouldn’t answer my questions on what the typical protocols are that people are using with grey market peptides, but Grok would… so here it is.

Please let me know where you think its missed anything, or is wrong:

Groks’ response:

Typical Processes and Protocols for Using Grey Market Peptides

Grey market peptides, such as BPC-157, TB-500, and semaglutide analogs, are often purchased as lyophilized (freeze-dried) powders from online research chemical vendors. These are typically labeled “for research use only” and not approved for human consumption in most jurisdictions, but users commonly reconstitute and self-administer them for off-label purposes like injury recovery, muscle growth, or weight management. The processes below are compiled from common user-reported practices and guides, which emphasize sterility to minimize risks like infection. Protocols vary by peptide, goal, and individual, but the core steps are similar. Dosages are highly variable and depend on factors like body weight and intended effects—users often start low (e.g., 250-500 mcg for BPC-157) and titrate up based on tolerance.

Always consult a healthcare professional before use, as these substances can have side effects and interactions. The following outlines the step-by-step processes, starting from receipt of the product.

1. Receipt and Initial Storage

  • Upon receiving the package (typically discreetly shipped in vials), inspect for damage or tampering.
  • Store lyophilized peptides in a cool, dark place: room temperature for short-term (up to a few weeks) or in a freezer at -20°C or colder for long-term stability. Avoid light exposure, which can degrade sensitive residues like tryptophan.
  • For peptides prone to oxidation (e.g., those with cysteine or methionine, common in TB-500), store in an oxygen-free environment if possible, such as with desiccants or under nitrogen.

2. Preparation for Reconstitution

  • Gather all materials (see complete list below).
  • Work in a clean, well-lit area: Wash hands thoroughly with soap and water, then wear disposable gloves to avoid contamination.
  • Allow the peptide vial and reconstitution solvent to reach room temperature if stored cold—this prevents condensation and shock to the powder.
  • Calculate the desired concentration: For example, for a 5 mg vial of BPC-157, adding 2 mL of solvent yields 2.5 mg/mL (or 250 mcg per 0.1 mL). Use online calculators or simple math: (total peptide mg / solvent mL) = concentration mg/mL.

3. Reconstitution (Mixing the Peptide)

This step dissolves the powder into a usable solution. Use bacteriostatic water (BAC water, containing 0.9% benzyl alcohol) for multi-dose stability; sterile water is an alternative but limits shelf life to 24 hours.

  • Step 1: Disinfect the rubber stoppers on both the peptide vial and solvent vial with alcohol swabs. Let air-dry.
  • Step 2: To equalize pressure (prevents spraying), draw air into a mixing syringe (e.g., 1-3 mL) equal to the solvent volume. Insert into the peptide vial and inject the air (or let vacuum pull it).
  • Step 3: Draw the calculated volume of solvent (e.g., 1-2 mL) from its vial using the mixing syringe.
  • Step 4: Insert the syringe into the peptide vial at a 45-degree angle. Slowly inject the solvent down the inner wall of the vial—avoid direct contact with the powder to prevent foaming or degradation.
  • Step 5: Gently swirl or roll the vial between your palms until the powder fully dissolves into a clear solution. Do not shake vigorously, as this can denature the peptide.
  • Step 6: Label the vial with the reconstitution date, concentration, and peptide name.
  • If the peptide doesn’t dissolve easily (e.g., neutral-charge peptides), consider dissolution methods based on charge: For positively charged peptides, use acidic solvents like acetic acid; for negatively charged, basic like ammonium bicarbonate; for neutral/hydrophobic, organic solvents like DMSO in small amounts before diluting with water.

4. Storage of Reconstituted Peptide

  • Refrigerate immediately at 2-8°C (36-46°F) in an upright position, away from light (e.g., in the original box or a light-proof container). Do not freeze, as this can cause precipitation.
  • Shelf life: Up to 28-90 days with BAC water; discard if cloudy, discolored, or past expiration. Minimize freeze-thaw cycles for aliquots if storing long-term.
  • For travel or extended use, some users aliquot into smaller sterile vials to avoid repeated exposure.

5. Dosing Calculation and Preparation

  • Determine your dose based on research or community protocols (e.g., BPC-157: 250-500 mcg daily; TB-500: 2-5 mg twice weekly; semaglutide: 0.25-2.4 mg weekly, titrated).
  • Use an insulin syringe for precision: Convert dose to volume (e.g., for 2.5 mg/mL concentration, 500 mcg = 0.2 mL or 20 units on a 100-unit syringe).
  • Optionally prefill syringes for convenience and store in the fridge in a light-proof container.

6. Administration (Injection)

Most grey market peptides are administered subcutaneously (SubQ) or intramuscularly (IM); some like BPC-157 can be oral but less common due to lower bioavailability.

  • Step 1: Clean the vial top and injection site (e.g., abdomen, thigh, or near injury for localized effects) with alcohol swabs.
  • Step 2: Draw the dose into an insulin syringe: Insert needle, invert vial, pull plunger to the mark, tap out air bubbles.
  • Step 3: For SubQ: Pinch 1-2 inches of skin/fat, insert needle at 45-90 degrees, inject slowly, hold for 3-5 seconds, then withdraw.
  • Step 4: For IM (e.g., BPC-157 for muscle repair): Insert straight into muscle (e.g., deltoid, glute) at 90 degrees, inject, hold, withdraw.
  • Step 5: Dispose of the needle immediately in a sharps container. Rotate injection sites to avoid irritation.
  • Cycle protocols: E.g., BPC-157/TB-500 combo: 250 mcg BPC + 2 mg TB twice weekly for 4-6 weeks, then maintenance or off-cycle.

7. Monitoring and Disposal

  • Track effects in a log: Dose, time, site, side effects (e.g., nausea for semaglutide).
  • Dispose of waste safely: Sharps in puncture-resistant containers; check local regulations for drop-off (e.g., via FDA guidelines).
  • If issues arise (e.g., infection, allergic reaction), seek medical help immediately.

Complete List of Materials Needed

Use the table below for a comprehensive list, sourced from typical setups. Quantities vary by usage volume.

Category Items Purpose Notes
Core Reagents Lyophilized peptide vial (e.g., 5-10 mg) The active substance Purchased from vendors; check purity certs if available.
Bacteriostatic water (BAC water) or sterile water (10-30 mL vials) Solvent for reconstitution BAC preferred for preservation; sterile for single-use.
Measuring Tools Insulin syringes (29-31 gauge, 0.3-1 mL, 100 units) Dosing and injection Fine gauge reduces pain; units for precise mcg measurement.
Mixing syringes (1-3 mL, 18-22 gauge) Drawing solvent Larger for initial mixing.
Micro-pipettes (optional, 10-1000 µL) Ultra-precise dosing Used by some for lab-like accuracy, especially with small volumes.
Sterility Items Alcohol swabs (70% isopropyl) Disinfection Essential for vials and skin.
Disposable gloves (latex or nitrile) Handling Prevents contamination.
Soap and water Hand washing Basic hygiene.
Storage Refrigerator (2-8°C) Post-reconstitution storage Standard home fridge; avoid freezer compartment.
Freezer (-20°C or colder, optional) Long-term lyophilized storage For unused vials.
Light-proof containers or original boxes Protection from light Aluminum foil or dark bags as alternatives.
Safety/Disposal Sharps container (puncture-resistant) Needle disposal FDA-cleared; available at pharmacies.
Needle clipper (optional) Safe needle removal For home use.
Miscellaneous Clean counter/workspace Preparation area Disinfected surface.
Labels and marker Vial annotation Date, concentration tracking.
Logbook or app Monitoring Record doses and effects.

This covers the end-to-end process as typically followed. Variations exist (e.g., oral capsules for some peptides require additional encapsulation tools like empty gel caps and fillers), but injection is the most common route for grey market use.

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I’m going to start tirzapetide. The vendor is trusted, but I am torn between getting vials of 30 ml or 10 ml, the former being much cheaper per weight unit but needing more punctures and more prone to contamination.

Anyone use 30 ml vials with no problem and can walk me through their sterilization / reconstitution protocol? @Steve_Combi ? Anyone? Or play it safe with 10 ml vials?

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I’m wondering about this also. Can’t you just repackage the larger 30ML (or larger amounts) into smaller vials or even sealable packages?

I think you guys mean 30mg, and not 30ml. When talking about dosage, no one talks about ml or units since that can vary based on the amount of bac water used.

You can, although with proper aseptic technique and filtering, your solution should stay sterile for 30 to 60 days.

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I’m talking about the original source powder… the lyophilized peptides. When you get the vial can’t you just split it up into smaller amounts? (pre solution).

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I also highly recommend filtering after reconstitution. Lyophilized peptide vials can be sterile, but that’s absolutely not a requirement. Here is a video that shows the whole reconstitution process with filtering

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No, absolutely not. You risk introducing bacteria. You also assume that you’ll be able to separate 30mg of peptide evenly, when most of the powder is filler.

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@qBx123Yk So what do you suggest? And yes you’re right it’s 30 mg of course not ml. Do you think it’s too big an amount? I am titrating up from nothing so it will take me who knows how long to even get to 7mg a week. I love me a bargain but I’m not sure it’s worth it.

I think 10mg is a good choice if that’s you want to go with. You don’t have or worry about poking your vial too much, and stay around 30 days of using the same reconstituted vial.

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Best thing is, this is not rocket science. Work in a relatively “clean” environment, not out in the barn, the kitchen is fine :slight_smile: , wash your hands before, and that pretty much does it for “sterilization”. 100’s of millions of people have been self injecting at home for decades, it’s very safe and easy.

Filtering is a waste of time. The most harmful thing that could possibly be in a peptide is an endotoxin and filtering will NOT remove that. Again a LOT of people are doing this with zero filtering and no issues. Paranoia will destroy ya :slight_smile: I’ve never filtered and I’ve been doing this for 2 years, with up to 10 injections in one day of different peptides. Not to mention all my family and friends, My average daily is 5 injections.

I keep all my vials (when I used to use vials) in a baggy in the fridge. Keeps random airborne stuff off my vials.

You don’t need much in the way of “sterilization”, just alcohol swabs. Wipe the top of the peptide vial before every insertion of a needle. When you draw Bacteriostatic water from the BAC water vial, wipe the top of that first as well. Wipe the injection site with an alcohol swab before injection, let it dry a bit or it may sting :slight_smile:

I use 2 different sized needles,

  1. for reconstitution - 5.0mL with a 22ga needle
  2. for injection - 1.0mL insulin syringe 29 to 32ga needle, marked off in “units”, 100 units = 1.0mL

I try to reconstitute with enough BAC water so my smallest injection is 10 units = 0.10mL If you don’t use enough BAC water the injection volume will be too low for accuracy.

Keep in mind all these vials are air tight. You must inject the same amount of air into the vial as you will draw out. With the BAC water vial, you would draw the plunger back to say 2.0mL Then insert in the vial and push the air into the the vial. This will make it much easier to draw out the 2.0mL of BAC water you need. This also applies to the reconstituted vial, if the injection volume is 50 units, draw 50 units of air into the syringe before and then inject that into the vial and draw out the 50 unit shot.

First step is to decide on the weekly dose.
This will tell you how long a vial will last.
10mg vial using a dose of 1mg per week would last 10 weeks.
10mg vial using a dose or 2.5mg per week would last 4 weeks.
With GLP1’s you can use a vial for at least 8 to 10 weeks without any issues.

Second step is to decide on how much bacteriostatic water to use. This is somewhat dependent on the volume of the vial. If they are 5mL vials, easy to deal with, 3mL vials also work well for GLP1’s

Third step is to decide on the volume of the shot. For GLP1’s I like to use more BAC water than less. This makes the shot more accurate.

In the case of a 10mg vial with a 2.5mg weekly dose I’d use 2.0mL of BAC water.
You would then have a shot size of 0.5m: (50 units on an insulin syringe (x 4 = 2.0mL BAC water)

Attached is a spread sheet you can use to play around with vial sizes, shot sizes and doses.

Reconstitution_Basic.xlsx (11.4 KB)

What @Steve_combi says :100::100::100:.

Buy economically, so 30 ml vials. Pay attention to your body’s response. If no weight loss in 2 weeks up the dose a bit. People with poor genetics seems to need higher doses to start the process.

Good luck!

Don’t pre-dispose yourself to some random dose you think you need, i.e. 7mg

The scale is your friend, if you are doing this for weight management, weigh yourself at least 3 times a week. I would not increase a dose mid month cycle, I would be patient and only increase dose after 4 weeks of weight data.

The clinical process is that the first month is the acclimation dose, it’s not a weight loss dose. You may lose weight or you may not, that is the process, let your body become acclimated to this new reality. GLP1’s are a significant change to how your body works. Let it adapt, there is no hurry.

After the first month asses your weight, if you have lost a few lb’s increase the dose a bit, not to the next clinical level, unless you want to deal with potential side effects. Low and slow will ensure success with very few side effects.

A reasonable dose progression is based on the scale (goal 1 lb per week) and goes something like this,

  1. month 1 - 2.0 or 2.5 mg per week for 4 weeks
  2. month 2 - depending on response and any sides 3.5mg to 4.0 mg per week x 4 weeks
  3. Month 3 - now you are fully acclimated, larger increase will have fewer sides now, again do not change dose mid 4 week cycle, be patient and see how it goes before increasing dose.

I have found that it takes longer that the 1 month acclimation to truly acclimate, 2 months minimum and for some it takes 3 or 4 months. Sure you can push this harder, as they do in the clinics (they need the result FAST so they don’t lose a client) but fast is not the best way for the client.

I’ve got more than a few who went to clinics that followed this faster protocol who dropped out due to the issues they experienced. With the low and slow concept I’ve not had one of those folks drop out as they are now successful with no sides.

This is either a lifetime of being at a healthy weight and deserves patience or it’s a vanity project that will be dropped as soon as it become difficult.

Avoid the difficulties and be successful :slight_smile:

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I had bookmarked a comment of yours with a titration schedule that’s similar to what you just mentioned (though I’ve bookmarked so many things I’m not sure I’d have found it :laughing:). But my question was about whether a 30 mg vial is safe to use. You haven’t directly answered that question but I suppose you did answer it indirectly: I should be dealing with very small doses for the first 2+ months so 10mg vials are the way to go as if I go as slow and steady as you suggest that 30 mg will last TOO long for safety.

I only need to lose about 20-25lb if that makes a difference.

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@Steve_Combi I’ve been doing more learning this month and it was only this week I saw the advice to inject equal amounts of air into the vial before you inject it with water or draw it up into the syringe… all this is to say that I’ve never done it and oddly have never noticed a problem…and with my cats, I’m drawing into a syringe several times each week. ??? Go figure

@qBx123Yk … I also use a filter… My theory is they are cheap and even if they most likely will never be needed, I figure why not. Cheap easy insurance.

And glad you posted a tutorial on venting… once upon a time, I didn’t know about venting, and a serious quantity of $$$$ alpha klotho sprayed all over my bathroom :). Painful!!! I don’t always vent my cheap stuff and I’ve never had a problem again, but I do try to remember to do it.

@RapAdmin as QB said, I would not risk contamination on splitting up larger bottles. I’ve seen you, so I’m assuming you are doing this for potential other health benefits vs weight loss. For that reason I think the lowest dose, and subsequently least expensive bottles would be perfect for you. Even without splitting it up, you can most likely take it small enough doses and maintain your weight. I initially lost 2-3 pounds on semaglutide which was great and I loved being less food obsessed. But after I added LDN, I lost more and then maintaining my weight started to become an issue (unlike glp, LDN killed my love of the taste of food, and even more of my obsession), so I just took even lower glp doses or less often. I have since used TZ and RT and find it is all very easy and manageable to control how much appetite suppression I want. I actually favor retatrutide because, while it’s nice that it also calms the noise down, I can still happily eat ‘through’ it.

If you by chance want to lose a couple pounds, I also think the smaller doses would still suit you best. If you wind up being a high responder and need to back off and waste product, I wouldn’t think twice about any of this going to waste because you can get a bottle of 15mg of TZ for $12.50! I personally have used a reconstituted bottle for up to 3 months (which QB reminds me I should not do), but even if you want to err on the side of caution and dump it after a month, you won’t worry about it.

@medaura I would defer to Steve, but I agree with QB that you should start at 10-15mg bottles. It’s all cheap enough that I think erring on potentially growing out of it is a better/safer plan. There is a good chance you will never need to go higher, but as you’ve heard, everyone responds just a little differently. Even if you wound up taking 10mg per week, unless my math is wrong, the $125 purchase of the 10 15mg bottles would still last you 3 months… so blowing through $125 in 3 months is pretty much the very worst case scenario that I don’t think is even likely to happen. And as Steve says, low and slow is the best plan and helps you with preserving muscle.

Also, @Steve_Combi helped my cousin lose 50lbs!!!

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A 30mg vial is perfectly safe to use, depending on the dose…

If the dose schedule uses up the entire vial in less than 8 weeks, it’s fine.

It’s not the amount of peptide in the vial, it’s the duration of use, which is determined by… the dose.

There seems to be two issues here you are bring up; the sterile environment, and the filler/distribution of the peptide in the powder.

We had a pharmacologist drop by here earlier talking about the sterility issue - regarding the similar issue of injections (different compound, but same approach I think). see here: Intramuscular (IM) + Intranasal (IN) Rapamycin - A new paradigm for human longevity translation - #124 by O_o

He recommended using a “still air box” similar to this design: How to Make a Still Air Box: The Complete Guide - Fungi Academy

The distribution of the powder is another issue. Does anyone have suggestions on how to get around this… do they even do this with a 3 or 4 gram order?

I’ve been talking with vendors regarding the purchase of 3 or 4 grams of SS-31, and it seems it would be about 100X more expensive if you purchase small dose vials (e.g. 10mg) than the full 3 or 4 grams.

Doing something that doesn’t work, is not insurance :slight_smile:

But if it makes sense to you, that’s all that really matters.

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I’m not trying to get endoxins that are too small to be filtered, but what about the very low chance of something bigger getting in there?

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I’d say low to none, at least in my 2 years of heavy peptide use.

But I know my supplier very well after doing business with them for 5 years.

If you don’t trust your supplier, why are you purchasing from them? Isn’t that why people do their “research”? to find the best and safest suppliers recommended by the other people we don’t know :slight_smile:

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I’m using and liking SS-31, so I’m intrigued by the price you are being quoted… also nervous about being able to handle that much raw volume on my own while keeping it sterile.

To handle that kind of volume, I think you are saying you’d potentially be using one of those boxes to be a mad scientist at your house?

I know most of the peptides you buy will be mixed with mannitol… but I also know it’s not necessary because Steve doesn’t use it (or at least he didn’t)… maybe he will comment. Having said that, when you aren’t using filler, you would need to be really really precise because the amount that would go into one vial for reconstitution would look like dust. SS-31 seems safe at high doses and a mistake might turn out to be fine, unlike puking one’s guts out on an OD of glp’s … ??

Keep us posted!!

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