Do You Filter Your Peptides Before Use?

[randompersonrandom]

Just wondering why he says he uses ultrapure water instead of sterile water for injection. 🤷‍♂️

Janoshik having a glass of peptides and getting ready to order some of the old Ultrapure.

 

[desinr-gal]

Janoshik wrote: "Sterility generally is not an issue with lyophilizates. There are not too many microbes that can survive either lyophilization or months while lyophilized. Those that can are not quite likely to find their way into the vials." That's from https://chat.peppys.org/t/lab-faq-guide-with-janoshik-analytical/4136. Click on the .pdf document and you can read the whole thing if you belong to Peppys.

Edited: I didn't realize that the quotation used strikeout type. I fixed that.

That is in regards to live microbes, there are other things that can be in there that maybe harmful. Jano has an entire business built on certifying these things are safe. He's not going to say yeah, we find all kinds of shii in there..

 

[keangkong]

I'm guessing that's sterile? 🤔

That is in regards to live microbes, there are other things that can be in there that maybe harmful. Jano has an entire business built on certifying these things are safe. He's not going to say yeah, we find all kinds of shii in there..

Janoshik will never certify anything is safe. He doesn't have the ability to test for everything.

 

[Nathanologist]

Ultra pure = no ions, organics, particles (chemically pure)
Sterile = no living organisms (biologically pure)

One is generally for lab work and the other is for injection. They are both essentially sterile, if they’ve been packaged in a sterile sealed vial.

 

[desinr-gal]

Janoshik will never certify anything is safe. He doesn't have the ability to test for everything.

No not literally.. Im just saying most of us use these certs, especially if there are more than one, to tell ourselves this is proof that they are safe, at least to an acceptable degree.
If people didnt place that much faith/weight on his coas, he wouldnt be so wildly successful.

 

[JoonyO]

50Shades got me to thinking about doing it just as a simple precaution. I ordered a couple of filter kits from P Test and now have ordered more of the doo dads I overpaid Ptest for from Amazon. Cheap peace of -mind even though I may be over-thinking the possibility of contaminants and my ability to filter them out. 🙄🫡

 

[Nathanologist]

Just push the plunger lightly and slowly.

 

[JoonyO]

Just push the plunger lightly and slowly.

Why? So I don't "push" the reconstituted pep out through the vent needle? Too late. Already did that. ☹️ Live and learn. 🤪

 

[Nathanologist]

Why? So I don't "push" the reconstituted pep out through the vent needle? Too late. Already did that. ☹️ Live and learn. 🤪

You can also blow out the filter. Ask me how I know… 🤦‍♂️

 

[JoonyO]

You can also blow out the filter. Ask me how I know… 🤦‍♂️

🤣 🤣 🤣
Oh great! Something else to look forward to! 🥳 lol

 

[desinr-gal]

You can also blow out the filter. Ask me how I know… 🤦‍♂️

So many times! urggg I am glad it's not just me!

 

[StonePny]

Why would he filter before testing? That completely defeats the purpose of purpose of the testing. I have never in my hours of research on janoshik of in my correspondence with the labs heard that stated or implied.

It was asked and answered of Jano 100% that he does filter before testing. It's to protect his equipment.

I assume "sterility" testing which is a separate test and has to be requested and paid for, is not done on the machines that are testing mass and purity.

 

[cgall88]

Twenty-eight days is the general guidance. I go up to six-weeks, I've read others do three months and longer. My opinion is that these meds are cheap, so why take chances? You really have to decide what your own risk tolerance is.

Thanks. My reasoning was that I considered getting the 60mg vial (x qty 10) but at the start my dosage would be literally 2mg p/wk. I don’t want to tamper with the mixing process so was planning 6ml BAC with the 60mg raw. On that logic even on a ramp up 4 weeks 2, 4 weeks on 4 the constructed vial would have been used for 8 weeks and I would only have used 24mg so the 1 start point vial of 60mg would still have 36mg. Think the penny pinching is silly.

 

[juGGaKNot]

Apparently you are not supposed to filter peptides with GHK in them, does anyone know more about this ?

Adsorption to filter membranes Peptides aren’t simple molecules – they’re amphiphilic, with both hydrophilic and hydrophobic regions, and often carry a surface charge. Depending on the filter type (PES, PVDF, PTFE, nylon, etc.), peptides can bind to the membrane via hydrophobic interactions, hydrogen bonding, or electrostatic attraction. This leads to peptide losses of 5–25 %, especially for small, basic peptides like GHK-Cu, which has multiple positively charged amino groups. Sources: Atha & Ingham, J. Chromatogr. A (1981); Ahern & Klibanov, Science (1985) ⸻ 2. Metal chelate instability (GHK-Cu) GHK-Cu is a copper(II) chelate complex of Gly-His-Lys. When passed through a membrane filter: • Cu²⁺ ions can adsorb to the filter, especially PES or PVDF (both have weakly basic sites). • The chelating equilibrium may shift, causing partial dissociation of the copper complex. Result: loss of active GHK-Cu, visible color change (bluish → colorless), and reduced biological activity. Sources: Pickart & Thaler, J. Inorg. Biochem. (1973); Maquart et al., Biochim. Biophys. Acta (1993) ⸻ 3. pH shift and ionic microenvironment Filter membranes have extractable ions (sulfates, carboxylates, or amines). When solution first contacts the membrane, it can cause a local pH drift of ±0.2–0.4 units — sufficient to denature or partially unfold sensitive peptides such as BPC-157 or TB-500. ⸻ 4. No sterility gain for lyophilized sterile powders Properly manufactured peptide blends (like KLOW) are sterile lyophilisates, meaning: • They were already passed through a 0.22 µm sterile filter before freeze-drying. • The vial was sealed under nitrogen or vacuum in aseptic conditions. If you reconstitute with fresh BAC water, new sterile needle, and wipe both stoppers with alcohol, you’re maintaining sterility equal to lab-grade conditions. Filtering afterward adds no sterility benefit but introduces a new contamination risk. ⸻ 5. Shear stress and conformational impact During syringe filtration, the liquid experiences localized shear forces, especially in small-pore filters. This can subtly distort peptide secondary structures, notably in β-turn-rich sequences such as TB-500. It’s minor but measurable in lab settings. ⸻ Summary table Aspect Effect of filtering Consequence Membrane adsorption 5–25 % peptide loss inaccurate dosing Cu²⁺ dissociation breakdown of GHK-Cu complex loss of efficacy pH / ionic change local denaturation instability Sterility no actual improvement contamination risk Shear stress conformational strain reduced bioactivity ⸻ In short Filtering KLOW offers no biochemical benefit and carries several downsides — you risk losing active peptide, destabilizing GHK-Cu, and even reducing shelf life. If the vial was sterile and vacuum-sealed, the correct approach is simply: → reconstitute aseptically, → store refrigerated, → use new sterile needles each time.

I asked janoshik and they do filter before testing.

Opinions ?

 

[JoonyO]

Apparently you are not supposed to filter peptides with GHK in them, does anyone know more about this ?
I asked janoshik and they do filter before testing.

Opinions ?

🤔 Hadn't heard of this as even a possibility and if it's true I just ordered some sterile vials for the KLOW I ordered for nothing. lmao 😂
Until I hear a to the contrary position the article you quote seems to be an affirmation TO NOT FILTER KLOW 🫡

 

[Nathanologist]

🤔 Hadn't heard of this as even a possibility and if it's true I just ordered some sterile vials for the KLOW I ordered for nothing. lmao 😂
Until I hear a to the contrary position the article you quote seems to be an affirmation TO NOT FILTER KLOW 🫡

A 0.22um filter is 220nm. Ghk-cu is a molecule with an approximate size of 1nm-5nm and is stable. The mechanical force is unlikely to cause damage, the filter is unlikely to restrict or filter it out, PES media is chemically inert… there’s no reason to avoid filtering this peptide that I’ve found. Perhaps loss which can be minimized with higher dilution and certain filtration techniques.

 

[JoonyO]

A 0.22um filter is 220nm. Ghk-cu is a molecule with an approximate size of 1nm-5nm and is stable. The mechanical force is unlikely to cause damage, the filter is unlikely to restrict or filter it out, PES media is chemically inert… there’s no reason to avoid filtering this peptide that I’ve found.

The article seems to FIND reasons you haven't: "In short Filtering KLOW offers no biochemical benefit and carries several downsides — you risk losing active peptide, destabilizing GHK-Cu, and even reducing shelf life."
So for now even not knowing the article's source it still resonates an air of sound rationale behind a reason to not filter KLOW. ( If it was created properly ).
But hey, what do I know? I'm just a kindergartner in Peptide school. I'll believe most anything.

 

[desinr-gal]

A 0.22um filter is 220nm. Ghk-cu is a molecule with an approximate size of 1nm-5nm and is stable. The mechanical force is unlikely to cause damage, the filter is unlikely to restrict or filter it out, PES media is chemically inert… there’s no reason to avoid filtering this peptide that I’ve found. Perhaps loss which can be minimized with higher dilution and certain filtration techniques.

The article quoted seems to warn about the TB and BPC, not the GHK.
"When solution first contacts the membrane, it can cause a local pH drift of ±0.2–0.4 units — sufficient to denature or partially unfold sensitive peptides such as BPC-157 or TB-500."

 

[desinr-gal]

Apparently you are not supposed to filter peptides with GHK in them, does anyone know more about this ?



I asked janoshik and they do filter before testing.

Opinions ?

Thank you for sharing this!

 

[juGGaKNot]

I am asking, the text I quotes reads like AI so I am inclined to not believe it

 

[zpped]

Apparently you are not supposed to filter peptides with GHK in them, does anyone know more about this ?
Opinions ?

Why would you not. Ask whoever told you that to explain what the problem is?

 

[Nathanologist]

The article quoted seems to warn about the TB and BPC, not the GHK.
"When solution first contacts the membrane, it can cause a local pH drift of ±0.2–0.4 units — sufficient to denature or partially unfold sensitive peptides such as BPC-157 or TB-500."

OK so let’s reason on this with what we find with a quick scholarly article search. Instead of making assumptions on how far down the rabbit hole my search has gone. Now I’m not a chemist or scientist or even a holiday inn express guest. But we have some of the most complete online scholarly source libraries and the most powerful search tools we’ve ever had access to.

1) BP-157 is not a sensitive peptide, it is widely described in literature as a stable gastric pentadecapeptide because it is native to gastric digestive juices. If BPC-157 can withstand hours in an environment that is several pH units lower than a neutral buffer, it is highly improbable that a localized, temporary drift of a mere 0.2 to 0.4 units in a buffered preparation would be sufficient to cause denaturation or unfolding. (Seiwerth, S., et al. (2021). Stable Gastric Pentadecapeptide BPC 157 and Wound Healing. Frontiers in Pharmacology, 12, 627533; Sikiric, P., et al. (2014). The Stable Gastric Pentadecapeptide BPC 157 Pleiotropic Beneficial Activity and Its Possible Relations with Neurotransmitter Activity. Molecules, 17(4), 461–481)

2) TB-500 is typically used as the synthetic fragment Ac-LKKTETQ, consisting of only four to seven amino acids. Denaturation and unfolding refer to the disruption of complex structures, which are characteristic of large proteins. Small fragments like TB-500 lack the intricate three-dimensional folds that would be susceptible to denaturation by a minor pH change. TB4 is however fairly unstable from what I’ve read and this is why almost all research is done with the fragment TB500. (Synthesis and characterization of the N-terminal acetylated 17-23 fragment of thymosin beta 4 identified in TB-500, a product suspected to possess doping potential-Simone Espositoa, Koen Deventera, Jan Goemanb, Johan Van der Eyckenb, Peter Van Eenooa)

3) The premise that a pH drift will occur and damage the product is typically nullified by standard pharmaceutical preparation practices, which require the use of proper buffers. The pH drift from filtration is transitory and happens briefly as the media is wetted. Now, normally in high quality pharma peptides they use a buffered solution to handle exactly this type of drift… we are probably not getting that type of product. TFA used to purify the petite may add to the pH problem. (Guideline on the Development and Manufacture of Synthetic Peptides. European Medicines Agency (EMA), Draft Guideline) However, as described in the literature above BPC-157 and TB500 have low susceptibility to denaturing even without a buffer.

4) Not having considered that the material I’ve read is likely referencing higher quality buffered peptides, I revisited the susceptibility of unbuffered GHK-Cu and KPV to filtration and pH drift. GHK-Cu is
stable at pH 4 to 7.5, but dissociation occurs at pH below 4, so testing pH prior to filtration could show you if you have room for the drift without damaging it. Apparently any TFA residual could cause the reconstituted solution to have a very low pH (GLOW/KLOW burn anyone?) which means it could be damaged even before filtering. KPV also a small peptide so now worries about denaturing, but attains a positive charge at pH below 7 so if the filter media has a negative charge it could selectively increase loss due to absorption. (https://www.researchgate.net/figure...ys-Pro-diketopiperazine-formed_fig4_266679185 ; https://pmc.ncbi.nlm.nih.gov/articles/PMC5498804/ ) I guess it seems reasonable to conclude that filter would negatively impact KPV and GHK-Cu. GHK-Cu in an unbuffered lyophilized powder with TFA residuals is already damaged if the reconned solution pH is below 4.

Again I not an expert by any means, but I can research read and reason ok. Those are my findings so far.

 

[Nathanologist]

The article seems to FIND reasons you haven't: "In short Filtering KLOW offers no biochemical benefit and carries several downsides — you risk losing active peptide, destabilizing GHK-Cu, and even reducing shelf life."
So for now even not knowing the article's source it still resonates an air of sound rationale behind a reason to not filter KLOW. ( If it was created properly ).
But hey, what do I know? I'm just a kindergartner in Peptide school. I'll believe most anything.

It’s not an article. It’s a Reddit post with citations that do not directly corroborate its conclusions. But the thought that grey market peptides may be lyophilized with an emphasis on purity but not on stability (ie no buffers) was one I hadn’t considered in all of my reading. The post makes some interesting claims worth investigating. Primarily for me is pH testing my reconned solutions since there may be no buffer.

 

[zpped]

(GLOW/KLOW burn anyone?)

The GLOW burn is certainly not PH based as many people have tested and confirmed the PH before experiencing it. The most likely culprit is free copper in the solution, as evidenced by other members showing that the burn can be neutralized by mixing in GHK-basic that would bind to the free copper.

We've been PH testing reconstituted peptides for a long time and almost everything is fine excluding most NAD as it requires a substantial buffer to make bearable and only PGB makes it that way last I knew.

 

[Nathanologist]

The GLOW burn is certainly not PH based as many people have tested and confirmed the PH before experiencing it. The most likely culprit is free copper in the solution, as evidenced by other members showing that the burn can be neutralized by mixing in GHK-basic that would bind to the free copper.

We've been PH testing reconstituted peptides for a long time and almost everything is fine excluding most NAD as it requires a substantial buffer to make bearable and only PGB makes it that way last I knew.

Do you find the pH lands somewhere between 6.0-7.0? Do you have any evidence that the solution is buffered? From what I’ve read even GHK-Cu when buffered is not going to be pushed out of the 4-7.5 by filtering.