Janoshik Analytical laboratory testing services

[Kralteramon]

I'd be quite surprised if it was GH.

I'd suspect its more of a correlation rather than causative relationship.

Nothing other than sending it in for a regular test occurs to me.
If it comes back shit, it's likely there's connection, but it's shooting in the dark.

Thanks Jano, I think that might be the case as well (correlation). I want to rule out some stuff through bloodwork and experiments since the test is just as expensive as the 700iu dirt cheap HGH and the supplier is not here anymore to take responsibility or co-fund the cost. I don't want to pay a reasonable but high price to find out it was not the HGH after all. I might just toss it at that point..

 

[Ghoul]

What size filter is used to prepare a sample for SEC analysis?

 

[janoshik]

What size filter is used to prepare a sample for SEC analysis?

.22um, as is industry standard

 

[Ghoul]

.22um, as is industry standard

I saw references to .22 and .45 so didn't want to assume.

What I think is interesting is this means SEC results are representative of what an end user would be injecting after .22um filtration.

For larger particles there seem to be a lot of different secondary methods used to get a more complete, "orthogonal" measure of larger particulates.

In house or outsourced, is there one available that could be used to augment SEC? I've seen MFI which seems straightforward to me (literally numeric counts of 1um, 2um, etc particles) but I have no idea if this is practical at a reasonable cost. That fact it captures other "trash" from the vial could be very revealing.

Anything you can offer in this regard?

 

[readalot]

I saw references to .22 and .45 so didn't want to assume.

What I think is interesting is this means SEC results are representative of what an end user would be injecting after .22um filtration.

For larger particles there seem to be a lot of different secondary methods used to get a more complete, "orthogonal" measure of larger particulates.

In house or outsourced, is there one available that could be used to augment SEC? I've seen MFI which seems straightforward to me (literally numeric counts of 1um, 2um, etc particles) but I have no idea if this is practical at a reasonable cost. That fact it captures other "trash" from the vial could be very revealing.

Anything you can offer in this regard?

Another excellent job Ghoul. Unfortunately few will appreciate the implications of this. Similar to folks thinking the aggregate test was just "dimer". Best wishes.

 

[Ghoul]

Another excellent job Ghoul. Unfortunately few will appreciate the implications of this. Similar to folks thinking the aggregate test was just "dimer". Best wishes.

Obviously this is not a limitation of the labs we use, or the formidable capabilities of those who make their invaluable expertise available to the community like Jano, but the limitations they have to work within based on our demands and the resources we make available to them.

There is little doubt large particulates present many types of risks, whether inorganic like glass and plastic from contaminated vials, or the well documented hazards of large aggregates presenting a large surface area of epitopes capable of triggering many types of immune responses with negative consequences.

Pharma has been keeping a tight limit on these particulates for quite a while, but the FDA still had to give the slow learners a "wake up call" a decade ago as they continue to tighten down the max limits of these contaminants.

 

[janoshik]

I saw references to .22 and .45 so didn't want to assume.

What I think is interesting is this means SEC results are representative of what an end user would be injecting after .22um filtration.

For larger particles there seem to be a lot of different secondary methods used to get a more complete, "orthogonal" measure of larger particulates.

In house or outsourced, is there one available that could be used to augment SEC? I've seen MFI which seems straightforward to me (literally numeric counts of 1um, 2um, etc particles) but I have no idea if this is practical at a reasonable cost. That fact it captures other "trash" from the vial could be very revealing.

Anything you can offer in this regard?

If someone has stuff larger than 0.22um in their GH, they have quite some problem, as at that point it's visible by bare eye. You measure that stuff with turbidimetry, which we can also do.

We do SEC inhouse.

 

[readalot]

If someone has stuff larger than 0.22um in their GH, they have quite some problem, as at that point it's visible by bare eye. You measure that stuff with turbidimetry, which we can also do.

We do SEC inhouse.

People inject "cloudy" GH every day and don't care. Not much you can do once the user is that apathetic.

Bubbles? No problem, haha.

Thanks for your services.

 

[janoshik]

People inject "cloudy" GH every day and don't care. Not much you can do once the user is that apathetic.

Bubbles? No problem, haha.

Thanks for your services.

My pleasure.

turbidimetry should be a fast test, we can probably try it out sometimes

 

[Ghoul]

If someone has stuff larger than 0.22um in their GH, they have quite some problem, as at that point it's visible by bare eye. You measure that stuff with turbidimetry, which we can also do.

We do SEC inhouse.

Below 100um is sub-visible, That's over 400x the size of what's removed by a .22um filter. Beyond that, many of us have seen particulates form after initial clear reconstitution, sometime in just minutes, sometimes days. Surely that means sub visible particulates can form even more quickly, if, for instance, there's no PH buffer.

This is what was detected in pharma HGH in an experiment led by a world leading expert in protein drug formulation, before and after .22um filtration. That's a lot of crap above .22um. I don't think it's unreasonable to think at least some would prefer to remove that prior to injection, in the reasonable belief it reduces risk. Those particulates serve no useful purpose. Removal certainly doesn't worsen risk. iI'm sure others are fine with it, and that's ok.

This is a good opportunity to advance harm reduction, raise standards closer to pharma. Controlling particulates this size is the norm, for well documented good reasons, and not a fringe theory.

 

[janoshik]

Below 100um is sub-visible, That's over 400x the size of what's removed by a .22um filter. Beyond that, many of us have seen particulates form after initial clear reconstitution, sometime in just minutes, sometimes days.

This is what was detected in pharma HGH in an experiment led by a world leading expert in protein drug formulation, before and after .22um filtration. That's a lot of crap above .22um. I don't think it's unreasonable to think at least some would prefer to remove that prior to injection, in the reasonable belief it reduces risk. Those particulates serve no useful purpose. Removal certainly doesn't worsen risk. iI'm sure others are fine with it, and that's ok.

It's a good opportunity to advance harm reduction, and certainly not a fringe theory.

View attachment 321236

You talk about single particle visibility, but if you have a sufficient amount of .22 um particles, the turbidity is visible by bare eye.

 

[Ghoul]

You talk about single particle visibility, but if you have a sufficient amount of .22 um particles, the turbidity is visible by bare eye.

I have a visible turbidity study somewhere, I'm curious how much and how large aggregates are needed to become cloudy to the naked eye. Beyond that, I've seen visible fibrils, which must be absolutely enormous, form in a short amount of time.

I'm seeing a theme in many of the studies on peptide degradation:

Large amounts of particulates are formed after some kind of stress, ie, PH, heat, shock. bubbles. but as they're mostly larger than 2um, not detected by SEC, but are by other methods.

For instance after dropping a vial of rHGH:

SEC:



MFI:



(on a side note, I've seen these gelatinous spots as well, and others I've discussed this with have too)





Anyway, I think many of us would appreciate a way to measure this important metric of quality and safety(a fun debate to be had with some "it's fine just pin it" dinosaurs I'm sure) , and would appreciate your efforts to be the first end user accessible lab to make it available.

DO NOT DROP: MECHANICAL SHOCK IN VIALS CAUSES CAVITATION, PROTEIN AGGREGATION AND PARTICLE FORMATION - PMC

Industry experience suggests that g-forces sustained when vials containing protein formulations are accidentally dropped can cause aggregation and particle formation. To study this phenomenon, a shock tower was used to apply controlled g-forces to ...

pmc.ncbi.nlm.nih.gov

 

[Ghoul]

Example of rHGH fibrils from a user here:

 

[AlexDavis43]

So we have a study showing aggregates in GH don't increase immunogenicity (GH antibodies) and GH antibodies don't decrease GH efficacy (ref).

There are also reports from people on Serostim, which has more aggregates than UGL hGH, who swear it is better than UGL hGH.

And some of the best IGF1 responses to GH on Meso came from one guy who used vacuumless GH and another guy who stored lyophilized GH at room temperature. Neither filtered.

 

[Ghoul]

So we have a study showing aggregates in GH don't increase immunogenicity (GH antibodies) and GH antibodies don't decrease GH efficacy (ref).

There are also reports from people on Serostim, which has more aggregates than UGL hGH, who swear it is better than UGL hGH.

And some of the best IGF1 responses to GH on Meso came from one guy who used vacuumless GH and another guy who stored lyophilized GH at room temperature. Neither filtered.

Each one of those statements is a gross mischaracterization, or a worthless anecdote.

 

[chumpsickel]

So we have a study showing aggregates in GH don't increase immunogenicity (GH antibodies) and GH antibodies don't decrease GH efficacy (ref).

There are also reports from people on Serostim, which has more aggregates than UGL hGH, who swear it is better than UGL hGH.

And some of the best IGF1 responses to GH on Meso came from one guy who used vacuumless GH and another guy who stored lyophilized GH at room temperature. Neither filtered.

So what you are saying is vendors should stop applying vacuum to the GH, and just buy the cheapest GH we can find. No point in testing because it might actually be better if the results suck. Got it.

 

[AlexDavis43]

So what you are saying is vendors should stop applying vacuum to the GH, and just buy the cheapest GH we can find. No point in testing because it might actually be better if the results suck. Got it.

Absolutely keep testing it to make sure you're still injecting GH.

Now, re-read what I wrote:

So we have a study showing aggregates in GH don't increase immunogenicity (GH antibodies) and GH antibodies don't decrease GH efficacy (ref).

There are also reports from people on Serostim, which has more aggregates than UGL hGH, who swear it is better than UGL hGH.

And some of the best IGF1 responses to GH on Meso came from one guy who used vacuumless GH and another guy who stored lyophilized GH at room temperature. Neither filtered.

 

[Ateam2023]

Example of rHGH fibrils from a user here:

View attachment 321257View attachment 321258

didn't you already post this somewhere?

 

[Ateam2023]

Each one of those statements is a gross mischaracterization, or a worthless anecdote.

just like you posting "essential meds list" ,, Why did you even say you would post it on more than one occasion if your just going to ignore the multiple requests from other members, , that's lame,, wrong thread didn't mean to clog up Janos thread, my apologies,,

 

[Ghoul]

So what you are saying is vendors should stop applying vacuum to the GH, and just buy the cheapest GH we can find. No point in testing because it might actually be better if the results suck. Got it.

An interesting thing about oxidation damage to rHGH. The study showed that it didn't directly damage the rHGH molecule.

What it did do was make the rHGH much more temperature sensitive, speeding up degradation, leading to aggregation.

Since more rHGH is pulled onto the aggregate, there's loss of active ingredient.

Once this aggregate becomes larger than .22um, it's no longer detected as a contaminant bringing down purity or measured as a loss of rHGH, since there's no way to know how much was in the vial to start with.

So the loss of purity you see in the test, and the huge increase in dimer, is only the tip of the iceberg. There are impurities and lost rHGH in the other side of the filter.

On top of this, the remaining rHGH will degrade faster than it should since it's been oxidized.