HCG degradation testing after 30, 45, 60 days

[Ghoul]

We have 2 data points:


Sample...Manufacturer....Excipients.....Reconstitution....30-day degradation
1................Liska.................?................Sterile water.............18.1%
2................QSC..................?.................BAC water................0.9%


Now the question is: Does the BAC water lead to reduced degradation, or the presence of (PH-stabilizing) excipients that may or may not be in the QSC product?

For those who think it's the BAC water: What's the mechanism here? The BAC water prevents the growth of airborne bacteria that otherwise would eat up the peptide hormone? @janoshik, what's your preferred explanation?

Of course BAC water delays degradation. Alcohol is a preservative, IE, creating an environment in which bacteria can not reproduce.

Pharma HGH reconstituted with sterile water is specified for single use, within 24 hours. Reconstituted with BAC it's good for 14 days and multiple uses.

https://pdf.hres.ca/dpd_pm/00004349.PDF

Medicines reconstituted with sterile water are never suitable for multi-dose vials.

It's worth noting if you fail to maintain proper technique when piercing that vial on a daily basis, failing to sterilize the stopper, especially if leaving it exposed in your refrigerator where circulating air will deposit bacteria on it. you'll be introducing more with each puncture, and speed up degradation. It should be kept protected in some container, and allowed to come to room temp before removing the vial so it doesn't collect bacteria laden condensation on it.

 

[MyNameIsJeff]

There is actually one more data point from this study I mentioned earlier: The influence of different excipients and storage procedures on hCG (human Corionic Gonadotropin) as evidenced by spectrophotometry - Nature Precedings

Sample...Manufacturer........Excipients........Reconstitution....30-day degrad.
1................Liska.....................?..................Sterile water.............18.1%
2................QSC......................?...................BAC water................0.9%
3................Pharma.......phosphoric acid.....Sterile water.............~0%
......................................&sodium hydroxide.........................................
......................................&Mannitol.......................................................

Regarding sample 3:

"The obtained solution was filtered through sterile syringefilters of 0.22 microns (Minisart 16534 K – cellulose acetate)to guarantee solution sterility.4. The obtained solution was bottled in 10 cc. vials and the vialswere tapped with rubber tops and aluminum securityprecincts.5. All described operations were performed under laminar fluxconditions."

So that could explain why the HCG did not degrade despite the use of sterile rather than BAC water. Then the common denominator for a lack of degradation is either the removal of bacteria, or the impairment of their growth.

Take-home message: Always use BAC water when reconstituting HCG, or any other peptide that you will use for several days or more (like the 40IU GH vials). Very important results, thanks for the testing Tracy/QSC!

 

[Ghoul]

Regarding sample 3:

"The obtained solution was filtered through sterile syringefilters of 0.22 microns (Minisart 16534 K – cellulose acetate)to guarantee solution sterility.4. The obtained solution was bottled in 10 cc. vials and the vialswere tapped with rubber tops and aluminum securityprecincts.5. All described operations were performed under laminar fluxconditions."

So that could explain why the HCG did not degrade despite the use of sterile rather than BAC water. Then the common denominator for a lack of degradation is either the removal of bacteria, or the impairment of their growth.

Take-home message: Always use BAC water when reconstituting HCG, or any other peptide that you will use for several days or more (like the 40IU GH vials). Very important results, thanks for the testing Tracy/QSC!

A related discussion came up in the QSC thread. Peptide manufacturers recommend filtering after reconstitution when a sterile solution is desired. They universally reccomend a .2um filter size, low protein adhesion filter,. which implies all peptides can pass through that size filter while blocking most bacteria.

Another added benefit of filtering is reducing the amount of large protein aggregates, which form in the presence of silicon and other contaminants often leaching from the container, especially the stopper.

These aggregates are known to cause an immune response, Some commercial therapeutic peptides specify they must be filtered at .2um prior to administration.

Of course this won't eliminate all aggregates, dimmer, etc, but fewer medical problems result when filtration is used.

Another reason to filter is that these aggregates can cause your system to develop antibodies to the peptide, potentially reducing its effectiveness in the future.

Probably should have its own thread but here's where it is now:

Post in thread 'Qingdao Sigma Chemical Co., Ltd (International, US, EU, Canada and Australia domestic'

Sep 22, 2024

Ghoul, are there any tests done on filtering peptides? Any chance of filtering the peptide and the drug being filtered out or reduced in the process? I have no idea how large the drug particle size vs mold/bacteria size. Not being a smart ass but I figured you would probably know. I read on another board about some peptides (I don't think they were qsc vials) sent in for testing failed sterility tests.

Yes. The major commercial peptide suppliers in the US advise .2 micron filtration can be used if sterilization of reconstituted solution is desired, which indicates, at least at...

• Ghoul

• 
• 

https://www.sciencedirect.com/science/article/abs/pii/S002235491830457X

PS: And yes, QSC sponsoring so much testing is a major benefit to the community, providing much needed hard data.

 

[Qingdao Sigma Chemicals]

Or could it be the difference between Urine purified and Recombinant HcG?

Urine purified hcg doesn’t exist anymore, let’s not bring this subject again.

 

[Qingdao Sigma Chemicals]

We can push the degradation test another 30 days if needed

 

[hungryalways]

We can push the degradation test another 30 days if needed

Yes, please! Or move to room temperature

 

[Crystal Clear]

Yes, please! Or move to room temperature

Do NOT move to room temperature unless start the process all over again with day 1 starting on room temperature.
QSC thanks for testing this

 

[mknodc13]

Urine purified hcg doesn’t exist anymore, let’s not bring this subject again.

Isn’t Eutrig-HP urine purified HCG?

 

[Qingdao Sigma Chemicals]

@janoshik was kind enough to offer to extend the test another 30 days, in the fridge, to see degradation in 60 days of HCG reconstituted, free of charge. Big thanks to him !

We will keep you updated once we have the results in a month.

 

[Qingdao Sigma Chemicals]

Isn’t Eutrig-HP urine purified HCG?

View attachment 296384

Possible.
I mean all HCG coming for China, the generic versions, there is no more piss versions, only recombinant.

 

[npbcs]

@janoshik was kind enough to offer to extend the test another 30 days, in the fridge, to see degradation in 60 days of HCG reconstituted, free of charge. Big thanks to him !

We will keep you updated once we have the results in a month.

Thank you to both Jano and Tracy for doing this.

 

[HUGE McBIGLARGE]

A related discussion came up in the QSC thread. Peptide manufacturers recommend filtering after reconstitution when a sterile solution is desired. They universally reccomend a .2um filter size, low protein adhesion filter,. which implies all peptides can pass through that size filter while blocking most bacteria.

Another added benefit of filtering is reducing the amount of large protein aggregates, which form in the presence of silicon and other contaminants often leaching from the container, especially the stopper.

These aggregates are known to cause an immune response, Some commercial therapeutic peptides specify they must be filtered at .2um prior to administration.

Of course this won't eliminate all aggregates, dimmer, etc, but fewer medical problems result when filtration is used.

Another reason to filter is that these aggregates can cause your system to develop antibodies to the peptide, potentially reducing its effectiveness in the future.

Probably should have its own thread but here's where it is now:

Post in thread 'Qingdao Sigma Chemical Co., Ltd (International, US, EU, Canada and Australia domestic'

Sep 22, 2024

Ghoul, are there any tests done on filtering peptides? Any chance of filtering the peptide and the drug being filtered out or reduced in the process? I have no idea how large the drug particle size vs mold/bacteria size. Not being a smart ass but I figured you would probably know. I read on another board about some peptides (I don't think they were qsc vials) sent in for testing failed sterility tests.

Yes. The major commercial peptide suppliers in the US advise .2 micron filtration can be used if sterilization of reconstituted solution is desired, which indicates, at least at...

• Ghoul

• 
• 

https://www.sciencedirect.com/science/article/abs/pii/S002235491830457X

PS: And yes, QSC sponsoring so much testing is a major benefit to the community, providing much needed hard data.

What’re the filters called exactly? Google ain’t helping me

Nevermind. I figured it out. Would freezing the peptides after reconstitution, and drawing from the sub-0c vial for use reduce the risk of proteins forming?

And how important is it to use a new vial after filtering? If I plunge the peptide back into the same vial, I’ve filtered maybe 98% of it. Does that reduce the risk by 98%, or does it not work like that? The reason I ask is the only vials I can get are UGL, so I’d rather minimise the contact the peptides have with UGL surfaces

 

[Ghoul]

What’re the filters called exactly? Google ain’t helping me

Nevermind. I figured it out. Would freezing the peptides after reconstitution, and drawing from the sub-0c vial for use reduce the risk of proteins forming?

And how important is it to use a new vial after filtering? If I plunge the peptide back into the same vial, I’ve filtered maybe 98% of it. Does that reduce the risk by 98%, or does it not work like that? The reason I ask is the only vials I can get are UGL, so I’d rather minimise the contact the peptides have with UGL surfaces

Filtering back into the same vial isn't ideal but it's better than not filtering imo. One thing I noticed is when the guys studying this do things to force aggregates to develop, it seems to take days for them to "incubate" and form, so fresh filtered would take a while to regenerate aggregates, even in the old vial.

 

[HUGE McBIGLARGE]

Filtering back into the same vial isn't ideal but it's better than not filtering imo. One thing I noticed is when the guys studying this do things to force aggregates to develop, it seems to take days for them to "incubate" and form, so fresh filtered would take a while to regenerate aggregates, even in the old vial.

And how about storing them in the freezer rather than fridge?

 

[Ghoul]

And how about storing them in the freezer rather than fridge?

The problem is ice crystals can cause physical damage to peptides.

The big peptide manufacturers say if you're going to freeze reconstituted peptide, put it into single dose size containers, so you only freeze/thaw once. Repeatedly cycling it in the freezer will degrade the peptide for sure.

Otherwise, low as possible but above freezing temps, help slow all types of degradation. Chemical. bacterial, aggregation.

 

[HUGE McBIGLARGE]

The problem is ice crystals can cause physical damage to peptides.

The big peptide manufacturers say if you're going to freeze reconstituted peptide, put it into single dose size containers, so you only freeze/thaw once. Repeatedly cycling it in the freezer will degrade the peptide for sure.

Otherwise, low as possible but above freezing temps, help slow all types of degradation. Chemical. bacterial, aggregation.

Thanks.

I’m gonna do this:
-Draw bact into a syringe. Plunge it into a peptide vial
-Draw the reconstituted peptide back into the syringe
-apply a glass fibre filter
-stick the reconstituted peptide back into the vial through the filter
-draw and inject from vial using insulin syringes

Won’t freeze it. Thanks again.

 

[clearheaded]

Urine purified hcg doesn’t exist anymore, let’s not bring this subject again.

uh? it 100% is majority of the pharma market in western world.

 

[Sampei]

What’re the filters called exactly? Google ain’t helping me

Nevermind. I figured it out. Would freezing the peptides after reconstitution, and drawing from the sub-0c vial for use reduce the risk of proteins forming?

And how important is it to use a new vial after filtering? If I plunge the peptide back into the same vial, I’ve filtered maybe 98% of it. Does that reduce the risk by 98%, or does it not work like that? The reason I ask is the only vials I can get are UGL, so I’d rather minimise the contact the peptides have with UGL surfaces

Post the link to the filters

 

[Ghoul]

Post the link to the filters

You need shipping to EU though right?

FYI, the ones used for aggregate removal (and sterilization) of peptide need to be:

-Material: PES

-Sterile

-Hydrophilic

-NON-Charged (if it doesn't specify that, it should say "low protein adhesion", "low protein loss", or "low protein adsorption".)

-.2 or .22um (or even .1um will allow peptides through and eliminate more aggregates, but really slow).

-Some recommend 4mm to minimize loss of liquid in the filter. I tried this and what I assume was "protein fouling" completely clogged it before 1ml got through. A good sign it's working. but not practical imo. 13mm is a good trade off. You can dilute your peptide a little more than usual to compensate, so the retained liquid won't cause as much a loss of active peptide). Pushing a syringe of air through the filter at the end will eject more liquid from the filter.

Expect a price between 40¢ and $3 depending on source and quantity.