"Generic" GH ASSAYS

[CensoredBoardsSuck]

Also go read the thread CBS is referring to. The retards couldn't even work out European way of writing dates and took it for a conclusive proof.

I did mix up the date system you use in Europe. Sorry if I'm not used to working with European standards. I'm used to working with the only standard that actually matters in the world - AMERICAN.

But at least I was willing to admit to my mistake. You still haven't admitted to fabricating data for David/Gentech/GETM, despite being caught red-handed.

 

[Dr JIM]

This wasn't geared towards you at all.

mands

Yea oops lol!

 

[Dr JIM]

Example?

He was paid for his services and he provided them for multiple sources there. Simec was also paid for their services to test the same products and failed to provide accurate results.

And what checks and balances have been instituted to ensure "Janos" one man band lab results are reliable? The LAB MANDS and I are using has well over 50 employees and runs some 600 protein samples MONTHLY

I mean so what "Jano bought an HPLC" but having an HPLC is entirely different than knowing how to USE IT!

 

[PrideofNYC123]

And what checks and balances have been instituted to ensure "Janos" one man band lab results are reliable? The LAB MANDS and I are using has well over 50 employees and runs some 600 protein samples MONTHLY

I mean so what "Jano bought an HPLC" but having an HPLC is entirely different than knowing how to USE IT!

When testing illegal substances, there's always going to be questions whether they're tested in Orange, Texas or Belgium.

But again, it's all we have. It comes down to who you do/don't trust and we can all decide that on our own.
Seeing your lab and Jano test the exact same samples would be interesting.


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[Eman]

Seeing your lab and Jano test the exact same samples would be interesting

Maybe... But seeing as how jano requires payment and mands and Jim are doing this free of charge I wouldn't put any faith in the results personally.

 

[Geterdun]

@jano know it all thread

Here you go @jano your very own thread to show us your studies that prove everything your claiming.

 

[CensoredBoardsSuck]

It seems Jane is having the same troubles at ProMuscle as he is here: He's full of shit. Lmfao

What YOU won't address Janoshik:

It doesn't matter what kind of "chromatograph" you use, they all do the same basic thing, and none of them will see tertiary protein structure.

It is perfectly obvious to me, and everyone here with a decent brain, that you are doing everything possible, including freaking out, posting in caps, spamming pointless information, to avoid this simple fact.

I should make a list of a hundred numbered bullet points with "why won't you address this point" on each one, but I don't care that much.

Yes it sucks we can't see tertiary protein structure with any reasonable method, yes it is one of the most frustrating things about GH, don't blame me though, I'm just the messenger. Deal with the reality, don't try to hide from it with other BS.

This is an attempt to throw so much text and info into a single post that is confuses the general reader and leads them to believe you know what you are talking about, when you do not. You are making a few HUGE assumptions that just aren't true.

The biggest assumption you are making is that a gas chromatograph can somehow see tertiary structure, it cannot. Below you will see what is required to see tertiary structure, read.

This page will tell you, and everyone reading this everything they need to know for the most part:

Protein Structure: Primary, Secondary, Tertiary, Quatemary Structures



THAT is what is required to see 3D structure, not a simple GC/MS test.

Further reading SPECIFIC to GH (I posted this years ago):

Structure-function studies on human growth hormone. Evidence that tertiary structure is essential for biological activity. - PubMed - NCBI

And:

Thermal stability: a means to assure tertiary structure in therapeutic proteins. - PubMed - NCBI

Now lets talk about what is REALLY going on that everyone should know about.

There are peptide synthesizing machines that are relatively cheap that are being used to make 191aa hGH. These chains sometimes fold on their own, but mostly get screwed up. Most of the GH being sold is this crap. It will test on the MS/GC as "real" GH, it will work in the body a little bit if some of it folds right on it's own. It will show up on a blood test as GH.

There are no machines that can make GH, only cells that are genetically modified to do so.

<keyboard drop>

Before I go into detail, let me say that this whole argument is from Janoshik not having a basic low level understanding of WHAT a chromatograph does. You cannot talk like you "know your shit" when you don't even have a BASIC understanding of WHAT a chromatograph is FOR. This guy doesn't even have ANY undergrad studies in biochem or molecular biology or he would understand exactly what I am talking about.

Further, to defend himself, he just tries to discredit rather than backup what he is saying.

He is desperate and childish in his posts. Personally, I find it disgusting.

So are you telling me that identifying changes in tertiary (and in cases quarternary) structure is impossible by any means other than most advanced analytical instrumentation?

Exactly. Any low level (undergrad) biochem student would know this. The fact that you DON'T know this means you shouldn't even be chiming in at all. It's actually a new development that we can see tertiary structure at all, even with "advanced analytical instrumentation."

I quoted the same wiki article (along many others) in my first reply to Kaladryn, but he failed to reply to any of the quoted references and pretty much all the direct questions aimed at him.

That along the obvious failure to grasp the concept of differences between various chromatographic methods makes me pretty sure he is uneducated on the subject.


I'm not sure what he gets out of spreading misinformations and plain lies (because if something is the truth you usually don't contradict yourself the same day, am I right?).


He was trying to make HGH serum testing make look unreliable for qualitative test (not the amount, just if it's HGH). Then he was trying to make IGF tests look like they can't provide information the members here are using them to provide. All that talking about magical peptides, without a single piece of evidence, just so much talk obscuring his lack of knowledge.

Now it's pretty much every other analytical method.


If I was a bit more paranoid I'd say he's somebody who really doesn't want anybody to have anything tested, by trying to make all the testing methods unreliable. If I was less paranoid I'd say Kaladryn is just a fool who gets a feeling of importance by trying to appear smart.

Even if those are not the cases, Mr. M96ss just proved him to be a liar and people like this should be disregarded.

Again with the vague personal attacks, trying to hide the fact that you don't even understand basic biochem.

All I'm saying is that GH bloodwork and/or GC/MS testing (or any other acronym of chromatography) is not sufficient. You MUST also see a rise in IGF-1 levels.

Why you want to refute this so hard is beyond me, all I can think is this entire thing is you trying to defend yourself from not understanding what a chromatograph is for.

 

[muscle96ss]

Seeing your lab and Jano test the exact same samples would be interesting.

Yes, and I am pretty sure we can all make this happen. To me this is the ideal scenario, you will have 2 vials from the same kit, with one vial going to each lab. Then we can make this totally transparent and have both labs agree to reveal all data and testing scenarios/methodologies. I think there is much that can be learned here.

 

[CensoredBoardsSuck]

Poor Jane was even corrected by Karl. That had to sting. lol

As always, I'd like to chime in and say, all these tests do is verify the correct aa chain (ie primary and secondary protein structure) they do not verify tertiary or quaternary protein structure, which is required for the molecule to activate the receptor, and is the major flaw in all GH tests, done by bloodwork or by GC/MS...

In simple terms: just because it is the right amino acid chain, and exact weight, doesn't mean it has the correct 3d structure. I know how disappointing and depressing this fact is, but it's true...

With all the respect, that is not true Sir.

1. HGH doesn't have quarternary structure.
2. Tertiary structure is the complex of secondary structures, so if those can be determined so can be tertiary structure.
3. Secondary and tertiary structure are quite important in interaction with enviroment (like GH receptor). Stationary phase in liquid chromatography (HPLC) is such enviroment and different structure of same amino acid chain causes different retention. Therefore it can be used for confirmation of 3D structure (resolution is imporant factor in this). Also the basis of SEC/GPC HPLC is separation by '3D structure.'
4. Regarding bloodwork/immunoassay the tests vary so much that it's impossible to tell in random case. Some antibodies are quite 'flexible' so they 'ignore' secondary etc structure or they cover only a small portion of the HGH molecule. Some antibodies will only bind to proper 3D structure.

GC can't really be used with proteins, only HPLC is used as far as I know for obvious reasons (GC works under high temperatures, which ain't really compatible with proteins).

Ya... this is why part of the USP basic quality control is the biological assay test (in addition to the chemical assay tests we're familiar with) - in which you test in vitro and in vivo activation of the GH receptors and actual effect.

It is not in EU pharmacopoeia, though, and I've never heard about issues of HGH not being active in EU preparations.

I surely do believe that non-biological methods are capable of determining the activity in vivo.

With HPLC there's the everlasting issue with getting the best resolution possible, but I think it could be possible with that method as well. Misfolded proteins are certainly separable if not by normal reverse phase HPLC then with size exclusion chromatography.

X-ray crystallography is certainly capable of that, but is costly and a great pain in the ass to work with.



I remember talking about biological assay with some Australian professor this spring, but we came to no conclusions unfortunately.

I would like to aplogise to Mr. Karl from Sciroxx and everybody here - I claimed that bioassays for HGH are not used in EU Pharmacopoeia as far as I knew, but going through the materials today I noticed it not to be true and that, indeed, bioassay is used in EU Pharmacopoeia, though possibly to a lesser extent than in US.

I have also gotten in contact with Eli Lilly - manufacturer of Humatrope and after they verify my qualifications I should get access to their quality assurance program data - ie. how does a legitimate pharmaceutical company test their HGH.

Yeah, Jane's a real expert alright. Lmfao

 

[CensoredBoardsSuck]

And what checks and balances have been instituted to ensure "Janos" one man band lab results are reliable?

Jane's "checks and balances" consist of hope. IOW, he's crossing his fingers and hoping it will be enough, but he's not quite sure and might need better equipment. He's still charging for his services though.

You can't make this stuff up. Fucking amateur!

Regarding the purity, I can only point out that I hope I haven't missed anything and that the next upgrade to my equipment will be stuff to be able to separate proteins better and on par with the strictest of the standards.

I was able to compare my process of purity testing with the processes that are used by different sciencists working in the field and with HGH (and here I would like to thank people who made that possible!!) and they were pretty much the same.

Simec adheres to this: http://docdro.id/kIC8ZE0 as far as I know. I used a process a bit different, (120Å column instead of 300Å, as my 300Å was, for some reason, providing worse performance, little bit different mobile phase composition etc). With the money I'll hopefully be getting from this I'll get a new 300Å column (500-600€) and stuff related.

 

[ickyrica]

Jane's "checks and balances" consist of hope. IOW, he's crossing his fingers and hoping it will be enough, but he's not quite sure and might need better equipment. He's still charging for his services though.

You can't make this stuff up. Fucking amateur!

This is all from PM? Thx...

 

[Geterdun]

Seems he is trying to find his own lab from the members pockets.

 

[muscle96ss]

A lot of misinformation here regarding jano. He is a good guy who is very passionate about what he does. Due to time constraints, I am not going to go into one of my long-winded posts about my personal experience with him. Just that I was highly impressed with his honesty, integrity, and knowledge; and I have complete trust in him.

 

[Sampei]

Well at least he is able to say: sorry I was wrong.
Not like a guy we all know well

Poor Jane was even corrected by Karl. That had to sting. lol











Yeah, Jane's a real expert alright. Lmfao

 

[Sampei]

Yes, and I am pretty sure we can all make this happen. To me this is the ideal scenario, you will have 2 vials from the same kit, with one vial going to each lab. Then we can make this totally transparent and have both labs agree to reveal all data and testing scenarios/methodologies. I think there is much that can be learned here.

This would be the best case scenario!

 

[Geterdun]

A lot of misinformation here regarding jano. He is a good guy who is very passionate about what he does. Due to time constraints, I am not going to go into one of my long-winded posts about my personal experience with him. Just that I was highly impressed with his honesty, integrity, and knowledge; and I have complete trust in him.

I don't know you well, but hope you come across with true intentions.

@jano come in to this thread fists raised and arms swinging. I haven't read one post with his name in it, until he showed up trying to talk his BS about how @Dr JIM and how @mands are giving bad testing information. No introduction, no resume, no credentials,(which is HIGHLY expected with the BS he is spewing) nothing. I'm in this thread to learn, not read how anyone thinks their so much smarter, that their testing is so much more reliable, how Dr J said this and Mands said that. I can read and have read every post in this thread more than once. I don't need @jano quoting their posts to bring conflict to this thread. I can read and I am sure there are others who read the entire post as well. Why can't he start his own thread, post his comments there if this thread is so wrong? There have been questions, this thread has gotten off track, but nobody, not even the HGH suppliers themselves have not come in this thread like @jano. Not even Frank and trust me, that's a big statement coming from me. Please help him start his own thread and let the community decide who they want to read.

 

[janoshik]

I believe there is a misunderstanding or I stated it wrong. We had a vial sent in from a member that only contained carbohydrates or starch. The AAA recognized there were no proteins present and then after testing it was determined it was starch.

Thank you for clarifying.

Well it would tell us exactly what was in the sample provided if it was indeed protein correct? Are you saying it would not identify the albumin protein percentage or otherwise?

I am pretty sure the quantitative information from AAA would be far from accurate, but I cannot claim so unless an experiment had been done.
But without a separation method beforehands I can't see AAA being as accurate as being on par with better suited methods.

I would think AAA would be better suited for identifying GH and much quicker? I believe HPLC would be a all around better choice for identifying HGH or other PED's. If you had a standard to go by? Am I assuming correctly?

mands

Yes Sir, absolutely. If you don't have a standard AAA and/or mass spectrometry would be the choice.

And what checks and balances have been instituted to ensure "Janos" one man band lab results are reliable? The LAB MANDS and I are using has well over 50 employees and runs some 600 protein samples MONTHLY

I mean so what "Jano bought an HPLC" but having an HPLC is entirely different than knowing how to USE IT!

Mr. JIM, you are correct pointing this out.

However, please mind that even SIMEC, with many employees screws things up for example, so that doesn't have to be all around argument. Also, with regulated substances I am sure that the regular quality assurance process in big commercial labs is almost non-existent.

Sorry for using SIMEC again as an example, but they were made to admit, that even though they claim the are accredited and adhering to GMP and GLP (and did claim they do so even for regulated substances!!) that they in fact are not accredited for testing such substances and don't adhere to standards of laboratory work.

I don't believe that you will find any laboratory doing so with questionable samples, maybe except DEA and antidoping agencies (and their equivalents). I think cooperating with them wouldn't prove to be really useful.

A lot of misinformation here regarding jano. He is a good guy who is very passionate about what he does. Due to time constraints, I am not going to go into one of my long-winded posts about my personal experience with him. Just that I was highly impressed with his honesty, integrity, and knowledge; and I have complete trust in him.

Thank you Mr. M96SS, I really appreciate your trust.

 

[Dr JIM]

Keep the commentary HERE I've no problem with it.

Hey Jano I noted a few of the HPLCS you conducted and cited on PM and you're asking me for more data, what a farce.

Im willing to bet his "testing" went something like this
Jano is given a Grey sample fully labeled to include the IU content

Jano then can adjust the HPLC device to match whatever quantitative value he is provided.

There is NO BLINDING involved, PM is happy and Jano is delighted as both will receive "fair compensation" sooner or later for work "well done".

Contrast this with what MANDS and I are doing in a fully BLINDED MANNER.

A holding mailbox receives unlabeled samples, that are then forwarde directed to the lab.

Until recently I never knew the manufacture or the sample concentration (yep some simply refuse to comply and sent a fully labeled Godtropin and Somastatin to the holding BOX, and both were trashed!)

The involved facility conducts the assays and forwards the results to me as a FULLY BLINDED certified lab, rather than a Jano one man lab band!

There simply is no comparison and why PM or ANY UGL is willing to accept such gross deviations from existing lab standards is beyond reason.

Hmmm but maybe it has something to do with the amount of MONEY involved!

 

[PrideofNYC123]

He received vials without their retail labels.

They were labeled/numbered in the same manner as yours.


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[Dr JIM]

Really, and thru what mechanism did Jano receive these samples?

Did he know he would be testing GH?

What was the standard he used for comparison?

Where is his concentration calibration curve?

Where on these HPLC does he correlate % with concentration in mg?

Where is the MW listed as compared to the standard?

Was a Mass Spec conducted to confirm the HPLCS MW estimate was accurate?

Did Jano ever confirm his HPLC assays were accurate and run an AAA on any of these samples?

ALL OF THIS INFO is standard for ANY protein based HPLC as it ensures reproducibility and reliability from one sample to the next?

To that end if you only knew the amount of information that was NOT included in Janos HPLCS you wouldn't even consider asking me for more "raw data".

Ya better finish cleaning the testing problem in your own back yard before ya try mine bc what we have posted is as good as it gets, excepting that which is required for published research.

Why don't you take a gander at my second attempt to post GH data using
HPLC and note ALL the criticism I received in spite of divulging much more than what this lab rat posted.

The bias was unequivocal and intentional.

Bottom line Jano gives them what they
WANT and I give them the evidence based TRUTH.

The FACT IS UGL GH remains a crap shoot, some a better crap shoot than others, and if your lucky maybe you will avoid STARCH being the cause of a sticky plunger.