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Were Your DIY Vials Properly Sterilized?

Updated July 17, 2023

Broadly speaking, probably not. Of the 4 different homebrew guides that were evaluated, 3 failed to offer adequate terminal sterilization and none provided any guidance on depyrogenation. None offered any form of quality assurance testing. Does that mean they’re unsafe? Not necessarily. Many, many trans people inject homemade HRT every single week. If it was categorically unsafe then there would be large reports of infections on popular DIY forums and discords. A brief meta analysis of academic literature indicates that when contamination is introduced to multi-dose vials, the preservatives in the vials often kill the contamination in under 48 hours. While these are not studies regarding pre-existing contamination, nor studies about HRT, we can assume that at least the same logic structure applies.

When we take that data, and we combine it with the consideration that the average person is likely capable of fighting off small amounts of bacterial/viral contamination from an injection, the picture becomes a bit more clear. There is always potential for complication, but most people do not experience it.

That said. The creators of most DIY injectables believe they are achieving terminal sterilization, which they likely are not. Their methods are misguided and not scientifically supported. It is our belief that the amateur pharmacist who is compounding these medications should have a stronger understanding of the process they’re undertaking, especially regarding the safety of their product. Relying on the patients immune system, and potentially a preservative in the vial, to keep them safe from a product you’re making is irresponsible. Especially when sterility doesn’t have to be totally out of reach to achieve.

HRT.cat recommends the following sterilization procedure for folks who do not possess special equipment or know-how.

How sterilization should be done at home

This is about as close as we can get to following compounding pharmacy guidelines while working out of our own homes and without investing in expensive equipment.

  • All glassware washed and sterilized/depyrogenated
  • All other tools autoclave sterilized
  • All work area surfaces and items wiped down with isopropyl alcohol 70%
  • Working in a makeshift cleanroom under a still air box or laminar flow hood
  • Using sterile syringe filtration (0.22um) to achieve terminal sterilization
  • Testing the integrity of the filter
  • Testing the product for contamination

Lena / Ultimate DIY

https://groups.io/g/MTFHRT/wiki/29602 - This is often held up as the most reliable DIY injectables guide, but unfortunately falls vastly short in terms of safe sterilization practices. Lena has been brewing DIY since the 90s, and honestly power to her for that. She has helped countless trans women gain access to hormone therapy both through her supplying it and through the influence she has had on the community. It is our belief that the majority of misinformation that plagues the DIY injectables community comes directly from Lena. Lena taught herself how to do this in 1990s Ukraine, resources weren’t exactly available on safe lab manufacturing practices for HRT. It worked for her and the information has spread. That said, times have changed and we deserve higher-quality information.

The guide has a single recommendation for sterilization: make your vials and then boil them in a pot of water for 30 minutes.

  1. 100°C is not hot enough to achieve any type of sterilization. This is considered dry heat sterilization, as we’re not applying steam to the HRT inside the vial. Dry heat sterilization requires a holding temperature of 180°C for 30 minutes (above the MCT oil’s smoke point).
  2. Nothing was sterilized before sealing the vials and nothing gets sterilized after they’re sealed.

Lena has a separate page that recommends using an autoclave. Those instructions are also invalid. We did a full write-up on putting sealed vials into an autoclave here.

In summary: don’t use this method. Don’t write a new guide based on this method. If these vials have any sort of sterility it is solely because of the benzyl alcohol included in the recipe.

Noire Labs

https://noirelabs.net/diy-estradiol-guide - This person probably got the bulk of her info from Lena’s guide but has supplemented it with a few additional safety practices.

She recommends 175°C for 60 minutes to sterilize the vials, after rinsing them in ethanol. She recommends cleaning your workspace with ethanol, love this! Then the instruction is to fill the vials and to boil in a pressure cooker for 30 minutes.

  1. 175°C for 60 minutes is adequate to sterilize glassware. However, as mentioned elsewhere, this is a holding time at 175°C, not a cooking time. It probably falls short of actual sterilization, though noble effort!
  2. Sterilizing vials is great! But they need to be depyrogenated as well. That goes for the rubber stoppers too.
  3. And finally, the terminal sterilization is to boil in a pressure cooker for 30 minutes. Because the vials are already sealed this is essentially an application of dry heat (steam can’t penetrate the vial). Even a high level of heat in a pressure cooker isn’t enough. More about this here.

Ultimately we are left with a semi-sterilized glass vial and no proper sterilization on anything else. We do not recommend any of this.

Otokonoko Pharmaceuticals / Lilian

https://archive.is/htP6g - This one really upset us because we know how loved and respected Otokonoko is. This is an archived version of her method that was since deleted. Someone had a backup.

The only method of sterilization mentioned is putting a flask in an oven at 230° (presumably Celsius) for 30 minutes to sterilize a container of HRT. This is bad for a few reasons.

  1. It doesn’t depyrogenate
  2. 30 minutes is likely just long enough to get a small-sized flask up to temperature. When sterilization guidelines state that you can heat something at 230°C for 30 minutes it is instructing you to get the solution to that temperature and then to hold it there for 30 minutes. This isn’t like when a cooking recipe says 30 minutes in the oven.
  3. Nothing has been done to sterilize the vials the solution will go into
  4. We don’t know what that type of heat will do to the hormone compound, we’re unable to find research
  5. There is tons of room for contamination when the mixture is transported to the vials.
  6. The smoke point of MCT oil is 160°C. There’s no way it’s a good idea to inject burned oil.
  7. The guide states that this is overkill on sterilization, and that a simple water bath “would kill most bacteria”

This guide doesn’t add up to proper sterilization in any way. Hopefully, Lilian has adopted better lab safety procedures since this guide was published in 2020, however without evidence of her improving her practices we wouldn’t order from Otokonoko.

Guide by Anon

https://files.catbox.moe/ax9efj.pdf - This guide circulates sometimes and can be found in comment sections around reddit. Likely elsewhere. It has much more info in it besides how to brew vials. It is by far the best guide we’d seen written until we encountered Tyger’s guides below.

Soak all glassware, stoppers, and caps in IPA 70% for 30 minutes, then autoclave for 15+ minutes. Sterilize misc glassware, it links to what at first glance appears to be good data on how to do that. Sanitize lab area. Optional gloves and still air box. Use a sterile syringe filter to perform terminal sterilization. Cap. Heat in oven at 275°F(135°C) for 6+ hours.

  1. IPA soak might be unnecessary, but it’s not hurting.
  2. 15 minutes isn’t an adequate amount of time in an autoclave.
  3. Still air box and sanitized gloves should be required, not optional.
  4. Lacks depyrogenation
  5. Once getting to temperatures as low as 135°C it is hard to validate if sterilization is taking place. Maybe someone who knows more than us can help calculate this. It’s also unclear if subjecting the hormone to that temperature for that length of time is causing it to break down some.

Overall this is a very good method considering the other ones We’ve evaluated. If we added in depyrogenation and some validation testing that the syringe filter worked we would rubber stamp it. This method is likely properly sterilizing vials but isn’t removing pyrogens/endotoxins.

Guide by r/steroids Wiki

#TODO - Guide is here

Manufactured by Tyger

Tyger’s Guides

We’re looking at their “medium-scale” guide v1.3

Tyger’s opening section: “Who can homebrew?” sets the stage for the rest of this guide. They suggest a series of qualities in a person that makes for a good brewers, statements we wholeheartedly agree with. Overall this guide is exceptionally smart and is laden with good advice.

This is hands down the best guide on how to brew HRT that we’ve read thus far. Obviously we like our guide more, but we’re going to be biased on that ;)

The medium scale guide is for bottle top filtering, a technique we would like to support on this site soon.

The small scale guide, is for a technique to purchase pre-sterilized empty vials and to push syringe filtered preparation straight into it. A viable technique for small scale brews (1-4 vials).

There are minor issues with these guides and techniques, but at a certain point some of this actually just comes down to opinion. These are likely THE BEST vials you can get in Europe, and they’re all made from a lens of mutual aid.

We are in active communication with Tyger to compare notes and in hopes of improving both their guides and our guide. The minor issues we have with Tyger’s guide aren’t listed here as Tyger is actively working to change certain parts of their published technique. Expect an even stronger v1.4.

Once again, if we were in Europe these would be our preferred DIY vials.

Vanna Pharma

Vanna Pharma does not list their brew technique on their website, but we sure wish they did! They use all the right buzzwords and they speak about their brew process with the attitude we want to see out of homebrewers: intelligence, humility, and a strong admission that this isn’t pharmaceutical quality.

If anyone finds Vanna’s brew guide, send it along.

Other Producers

If you find the brew guides of ANY producers that aren’t listed here, please send them to us! We’d love to review their methods to add to this page.