DISCLAIMER: This is not medical advice, engage in DIY HRT at your own risk; the
intent of this post is harm reduction. This information is not intended for
minors; by reading further you agree you are over the age of 21. DISCLAIMER 2:
This is not rigorous scientific research. I conducted this experiment over the
course of a work day, carrying out my actual work while trying to not arouse
suspicion. I did not have time to create a calibration curve to conduct
quantitative analysis. Further, isolating samples and using IR and NMR to
identify impurities would increase my chance of being found out by coworkers. I
also forgot to collect a data point and made another mistake that I explained in
the “results” section that I did not have time to correct. That being said,
these results are roughly accurate and I am using them to inform my own DIY HRT.
This is somewhat of a follow-up of this post on DragonOrdnance estradiol
enanthate [https://hexbear.net/post/1419643], and I plan on posting every year
with stability data for prepared solutions when properly stored (cool, dark,
dry). Since I have 10 vials with 7 mL each, I have enough to provide over a
decade of stability data. I also have leftover powder which I also plan on
testing yearly. Abstract: To improve the safety of the DIY transfem community,
it is necessary to demonstrate the thermal stability of estradiol esters to
ensure DIYers properly sterilize injection vials. Without an autoclave, a higher
temperature is necessary to destroy all bacterial endospores that may be
present. Estradiol enanthate (EEn) has considerable thermal stability and can be
sterilized at home without the fear of decomposing the EEn. At the suggested
home-sterlization conditions of 130degC for 30 minutes, no decomposition was
detected. Even at 180degC for 1 hour, less than 5% of the EEn decomposed into an
unidentified compound. Light exposure also has minimal effect, with no
decomposition detected after six months of continuous exposure to artificial
light and indirect sunlight. Home sterilization is recommended to avoid
infection. However, it is recommended to store both EEn powder and EEn solutions
in a cool, dry, dark area to minimize the decomposition of EEn over longer
periods of time (EEn powder is shelf-stable for five years and likely much
longer). A follow-up study will focus on long-term stability of properly stored
EEn powder and EEn injection solutions. Introduction: Last week, I saw a post
about a comrade who started HRT and mentioned their sterilization method
(130degC for 30 minutes). A discussion in the comments included users suggesting
that estradiol enanthate (EEn) would start to decompose at 130degC. This
surprised me, as I remember seeing somewhere that estradiol ester injection
solutions can be sterilized at higher temperatures than an autoclave (121degC)
without significant decomposition. This is important because autoclaves are
effective not only for the high temperature, but for the elevated pressure as
well (~30 psi, or ~2 atm). Therefore, slightly higher temperatures are required
to effectively sterilize injection vials. DIYers likely don’t have access to an
autoclave or any kind of pressure vessel, so to improve the safety of transfem
DIYers, I aim to alleviate concerns regarding the thermal stability of EEn. I
have a considerable stash of EEn powder that I purchased 1.5 years ago (likely
over 2 years past its manufacture date) and I have an injection solution I
prepared in bulk, originally intending to prepare vials as needed. This bulk
solution has not been subjected to heat but has been on my desk at home where
it’s been subjected to artificial light and indirect sunlight (light, like heat,
accelerates decomposition of EEn). In this experiment, I determined the
stability of EEn under various conditions: as a powder and a solution, and after
exposure of the solution to light and/or heat. Methods: All injection solutions
were prepared as 50 mg/mL EEn in MCT oil with 2% (v/v) benzyl alcohol. The EEn
powder, MCT oil, and benzyl alcohol were kept at room temperature in opaque,
resealable plastic bags since receiving them 1.5 years ago. The light-exposed
injection solution was kept at room temperature in a clear, resealable plastic
bag where it was continuously exposed to artificial light and indirect sunlight
for 6 months. Two more injection solutions were prepared the same day as this
experiment to represent injection solutions that have not been exposed to
significant light. The “light-exposed solution” was analyzed by GC-MS. Then, the
solution was sealed in injection vials and the vials were placed in a GC oven at
130degC for 30 minutes. This “light-and-heat-exposed solution” was analyzed by
GC-MS. One of the freshly-prepared solutions was sealed in injection vials and
placed in a GC oven at 130degC for 30 minutes. This is the “heat-exposed
solution”. The other freshly-prepared solution was transferred to an open test
tube and subjected to a 180degC oil bath for 1 hour. This is the
“high-heat-exposed solution” and represents the worst-case scenario of a DIYer
sterilizing at a temperature far beyond what is necessary. Results/Analysis: The
chromatogram of the EEn powder suggests this is a high purity sample, as the
only peaks are EEn (at 26.4-26.9 minutes) and the MCT oil from previous analyses
(Image 1) This EEn powder was analyzed following the analysis of several EEn
solutions, and the needle wash was empty, so there were three other peaks that
were confirmed to be triglycerides from the MCT oil residue on the needle. There
is also a solvent peak (acetonitrile) at ~1-2 minutes. The mass spectrum is
consistent with that of EEn (Image 2).
Show [https://hexbear.net/pictrs/image/8197eb29-f1cb-493b-8559-3ed5796eb142.png]
Image 1. GC Chromatogram of EEn powder
Show [https://hexbear.net/pictrs/image/631b836d-3cc8-41c5-9bbb-49e36dfa7363.png]
Image 2. Mass spectrum of EEn powder Image 3 demonstrates a newly prepared EEn
solution that was exposed to light and atmosphere while being heated at 180degC
for 1 hour. Samples were diluted by adding 1 gram acetonitrile for every 10 mg
of sample. As the samples were at the same concentration, the peak area of the
EEn peak is a reasonable (though rough) measure of the remaining EEn. Before
heating (t= 0), the peak area was 186.0 M counts. At t= 15 min, peak area was
184.9 M counts. At t= 30 min, peak area was 178.7 M counts. At t= 1 hr, peak
area was 195.4 M counts. The peak area at 1 hr had the highest z-score at z=
1.5324, giving a p-value of .1254. This indicates it is very likely that the
variations in these measurements are due to chance. The chromatogram at t= 1 hr
demonstrates the production of a decomposition product, but the peak area of
this decomposition product is <5% of the area of the EEn peak (Image 4).
Show [https://hexbear.net/pictrs/image/7e942fe5-3b10-4c65-a70c-a90741e430e6.png]
Image 3. High-heat solution chromatograms
Show [https://hexbear.net/pictrs/image/dd78a72e-c522-4350-b74f-22a9420c8943.png]
Image 4. High-heat solution chromatogram at t= 1hr, zoomed-in to visualize
decomposition product peak The following three chromatograms represent the
light-exposed solution that was not heated (Image 5), the light-exposed solution
that was heated at 130degC for 30 minutes (Image 6), and the newly-prepared
solution that was heated at 130degC for 30 minutes (Image 7).
Show [https://hexbear.net/pictrs/image/f2ad614d-e422-4c23-9f43-46fc4025a4cf.png]
Image 5. Chromatogram of EEn solution exposed to light for six months
Show [https://hexbear.net/pictrs/image/8cfb2b22-b108-4acb-b906-3e66c1871b77.png]
Image 6. Chromatogram of EEn solution exposed to light for six months and
130degC for 30 min
Show [https://hexbear.net/pictrs/image/50590f5b-6840-40da-a88a-333c44606ef8.png]
Image 7. Chromatogram of EEn solution exposed to 130degC for 30 min, but no
light exposure In absence of rigorous quantitative analysis, the proportion of
peak area of EEn to benzyl alcohol was used to compare the decomposition of EEn.
The concentration of benzyl alcohol in these solutions was constant, so it is
another reasonable yet rough estimation of remaining EEn to determine if
decomposition occured. For the light-exposed/no-heat solution, the EEn peak was
68.99% the area of the benzyl alcohol peak. For the light-exposed/heat-exposed
solution, the EEn peak was 68.41% the area of the benzyl alcohol peak. For the
no-light/heat-exposed solution, the EEn peak was 68.20% the area of the benzyl
alcohol peak. These values are not significantly different, indicating that
decomposition did not occur in any of these samples. (I forgot to measure the
peak areas of a no-light/no-heat solution, I’m sowwy :(, but you can look at the
180degC data to see that no significant decomposition occurred at 30 minutes at
such a high heat). Conclusion: According to GC-MS data, no significant
decomposition at normal sterilization conditions (130degC for 30 minutes). Even
at 180degC, EEn is fairly stable, remaining 95% intact after one hour at this
extreme temperature. Light exposure is also a minimal risk, as no significant
decomposition occurred after six months of moderate light exposure.
A user posted some incredibly good science on DIY HRT, I have verified them, they wish to stay anonymous.
Please take a look at their post! I know people have a vendetta against hexbear yada yada yada but please take a look at the post, it is very good information.
Tldr?
Sterilize at 130C is fine and good. Storage area and heat seems to have minimal impact on estradiol enanthate decomposing