"Generic" GH ASSAYS

[mands]

You know what I will do if people can't let it go and keep stirring the pot! I will make a private group and only invite whom @Dr JIM and I see fit.

That will take care of all the mud slinging

mands

Though there is a 'Steroid Lab Testing' section, personally I would rather have it under the 'Human Growth Hormone and Peptides' section. I don't consider rHGH a steroid and believe it is better suited/categorized here. IMO

That's my thoughts as well.

mands

 

[mands]

THE new "Generic" HGH Assay PAGE! AAA testing

mands

 

[janoshik]

With deference Millard, as we discussed CBSs idea would spread bt 50-100 samples over 50-100 threads and is just not workable as a means of evidence consolidation.

Although a spread sheet may provide a solution for single line narratives like those used by SIMEC on Anabolic Labs, such a format grossly deemphasizes the importance of graphic and worksheet analytical data from an evidence based perspective, as latter distinguish legitimate data from what Meso already has plenty of, unreliable nonevidence based "testing".

To that end, exclusive of a group forum or reporting results to individual donors via email or PM, the best option seems to be linked teferencing similar to what @Eman has posted as it enables members to access the original evidence in a fully transparent manner.

Jim

Really, let see the worksheet data I've posted on each sample on some spread sheet ?

Exclusive of the originators selected items a spread sheet primary purpose is to provide a summary of the RESULTS primarily, and while I agree that's what some want, it undercuts the validity of EVIDENCE based assays

A similar SIMEC narrative GH "spread sheet" report was recently posted on PM which lacked credibility bc the techniques and or methodologies were omitted Millard.

And that is the problem with spread sheets unless the original data can be referenced for credibility

Meso has a plethora of testing that's been posted over the years ranging from Lab Max "Angus" MS and little of it should be considered legitimate bc the results lacked credibility.

I am a fan of extensive data from testing , but why was it refused, numerous times, to provide such data in the first pages of the thread?

I think you said you don't want to overflow community with data, but I'm sure it would be useful for many, Mr. Dr. JIM, and for those for whom it would not be useful - those could just use the spreadsheet. Maybe somebody educated would come and help us all interpret the data, sir.

One of the FEW data related questions in this entire thread and is also one of the reasons those who rely on spread sheets exclusively as a measure GH quality are only cheating themselves.

So 4 grams of glycine made about 5ng raise in HGH blood levels. In fasting subjects.

The reaction seems to be dose dependent from the articles you have quoted.

4g of glycine 5ng HGH raise
8g glycine 9ng raise in HGH levels
12g glycine 16ng raise

So are you saying that the producers are adding 4 grams of glycine to vials with a goal in their mind to produce HGH levels raise?

I think 4 grams are the minimum, because if they only added 2 grams, it could be extrapolated as 2.5ng raise in HGH levels and that would be useless, right?

So are you saying, that the manufacturers are adding multiple grams of glycine into the HGH vials? I'd be seriously worried about them doing that!!!

 

[Dr JIM]

^^^^^

The articles I've cited only provide a reason why glycine
is either being added OR not removed from generic GH products anything else is CONJECTURE.

 

[janoshik]

So you are saying, that the miniscule amount of glycine (because anybody who knows jack shit realises than even half a goddamn gram of glycine in a vial is absolute nonsense and every single customer can BREAK a vial and weight the powder with 20$ scale to see that it's never over 100mg per vial) is being ADDED for a hope of what? 0.2ng raise in HGH? Do you yourself believe that?

Or that glycine is intentionaly not removed? Do you have any idea how purification process works? Do you have any idea how much more expensive and complicated it would be to purify BOTH HGH and GLYCINE , which have vastly differing properties? Do you realise that two separations would be necessary? For what? 0.2ng raise in HGH? Along with the risk that other low molecular stuff gets in along with glycine?

Absolute nonsense.

Why have you not sent the raw data yet if you hate the 'incomplete data' so much?

I would absolutely love to see them and interpret them. Coincidentally I kinda am involved in testing stuff. AAA as well. I sorta am MD working in research.


Also, please, tell me how is AAA MORE PRECISE THAN HPLC again.

Please, just tell me so. It would help to explain to WHOLE PHARMACEUTICAL INDUSTRY which is using HPLC as routine quality testing for PHARMACEUTICAL PRODUCTS.

Or is European and US pharmacopoeia written by noobs in your opinion?


I mean, don't get me wrong. TESTING STUFF FOR FREE FOR PEOPLE IS AN AWESOME THING TO DO AND I'M FULLY UP TO SUPPORT IT.


WHAT I DO NOT SUPPORT IS SPREADING MISINFORMATIONS AND BULLSHIT.

 

[Eman]

I sorta am MD working in research.

I'm an astronaut. Also, I'm a pornstar.

 

[RandallNowan]

I thought the hormone was too fragile to survive the HPLC process? HP being "high pressure", of course, and HGH being something 'almost living'...

 

[Sampei]

I'm an astronaut. Also, I'm a pornstar.

Bullshit your pecker can't go past 3inch! You are a taker not a giver.

Liarrrrr

 

[Eman]

Bullshit your pecker can't go past 3inch!

Psh.. it's all about the girth anyways Sampei.

 

[janoshik]

I'm an astronaut. Also, I'm a pornstar.

Thank you for a valuable point in discussion. You sure are gettin a pat on the back for that one!

I thought the hormone was too fragile to survive the HPLC process? HP being "high pressure", of course, and HGH being something 'almost living'...

No Sir, that is a misconception and an absolutely untrue. HPLC is used even with much more fragile molecules than HGH.

The pressure of the fluid the HGH is dissolved in has no effect on the structure. HGH is sucessfully separated by liquid chromatography (LC) at pressure well exceeding 600 bar while still retaining ALL biological function.

Routine HPLC analyses seldom go over 400 bar. UHPLC over 600.

When I get back home I can send you several articles discussing this if you are further interested. Pubmed articles.

 

[mands]

So you are saying, that the miniscule amount of glycine (because anybody who knows jack shit realises than even half a goddamn gram of glycine in a vial is absolute nonsense and every single customer can BREAK a vial and weight the powder with 20$ scale to see that it's never over 100mg per vial) is being ADDED for a hope of what? 0.2ng raise in HGH? Do you yourself believe that?

Or that glycine is intentionaly not removed? Do you have any idea how purification process works? Do you have any idea how much more expensive and complicated it would be to purify BOTH HGH and GLYCINE , which have vastly differing properties? Do you realise that two separations would be necessary? For what? 0.2ng raise in HGH? Along with the risk that other low molecular stuff gets in along with glycine?

Absolute nonsense.

Why have you not sent the raw data yet if you hate the 'incomplete data' so much?

I would absolutely love to see them and interpret them. Coincidentally I kinda am involved in testing stuff. AAA as well. I sorta am MD working in research.


Also, please, tell me how is AAA MORE PRECISE THAN HPLC again.

Please, just tell me so. It would help to explain to WHOLE PHARMACEUTICAL INDUSTRY which is using HPLC as routine quality testing for PHARMACEUTICAL PRODUCTS.

Or is European and US pharmacopoeia written by noobs in your opinion?


I mean, don't get me wrong. TESTING STUFF FOR FREE FOR PEOPLE IS AN AWESOME THING TO DO AND I'M FULLY UP TO SUPPORT IT.


WHAT I DO NOT SUPPORT IS SPREADING MISINFORMATIONS AND BULLSHIT.

"Generic" GH ASSAYS

Here is a page of data posted on the greys.

Isn't a AAA the most accurate and absolute qualification of proteins that are of smaller concentrations?

Why wouldn't it be for larger amounts as well or any amount for that matter?

mands

 

[janoshik]

"Generic" GH ASSAYS

Here is a page of data posted on the greys.

Isn't a AAA the most accurate and absolute qualification of proteins that are sub-picomolar?

Why wouldn't it be for larger amounts as well?

mands

Thank you Mr. Mands, I have read it - is it all the raw data available? What about the system used? Separation method used? How it was calibrated?


And no, it is not the most precise method. For quantification it can range up to 15% inaccuracy.

Also please mind that AAA is DEPENDENT on HPLC. HPLC is part of the AAA and AAA can be only as precise as the HPLC separatation used. If there is no separation (how I suspect it was in this case!) beforehands, the results become even less precise.

So in ideal case you have to run 2 HPLCs to get AAA data. So any inaccuracies MULTIPLY.

 

[mands]

Thank you Mr. Mands, I have read it - is it all the raw data available? What about the system used? Separation method used? How it was calibrated?


And no, it is not the most precise method. For quantification it can range up to 15% inaccuracy.

Also please mind that AAA is DEPENDENT on HPLC. HPLC is part of the AAA and AAA can be only as precise as the HPLC separatation used. If there is no separation (how I suspect it was in this case!) beforehands, the results become even less precise.

So in ideal case you have to run 2 HPLCs to get AAA data. So any inaccuracies MULTIPLY.

All this info can be provided. I will speak to Jim about it and see if we can link it in the hyperlink on the new thread.

I know this info is all hard data and it would have to be all scanned a posted because it is not in PDF format like the analysis.

Here is what I found going back looking at my notes:

"Amino-acid analysis has a long history in the characterization of protein-based products, since it provides information on the product concentration without referring to an external protein standard and it is independent from the shape and the charge of the protein. In addition, the determined amino-acid composition can confirm sample identity and gives a measure of sample purity. Furthermore, when combined with absorbance measurements, it allows the determination of extinction coefficients under various conditions.1 For protein conjugates, where the synthetic counterpart modifies the protein absorption properties, amino-acid analysis may be required as the only reliable quantification method.

However, in spite of these features, few laboratories can perform such analysis in a reliable and quantitative way, due to the need for specialized equipment and skills. Usually, techniques based on ion-exchange separation coupled with post-column derivatization (e.g., with nin-hydrin, the “classical” method) are considered more precise1 than those based on pre-column derivatization and reversed-phase high-performance liquid chromatography (RP-HPLC), because the latter techniques imply extensive sample manipulation before analysis and are affected by the limited stability of the preformed derivatives.2 However, such RP-HPLC-based methods have the advantage of being accessible to most analytical laboratories, since they do not require expensive dedicated instruments. In addition, manufacturing of dedicated instruments is being halted, making the availability of validated pre-column methods even more important."

mands

 

[mands]

Thank you Mr. Mands, I have read it - is it all the raw data available? What about the system used? Separation method used? How it was calibrated?


And no, it is not the most precise method. For quantification it can range up to 15% inaccuracy.

Also please mind that AAA is DEPENDENT on HPLC. HPLC is part of the AAA and AAA can be only as precise as the HPLC separatation used. If there is no separation (how I suspect it was in this case!) beforehands, the results become even less precise.

So in ideal case you have to run 2 HPLCs to get AAA data. So any inaccuracies MULTIPLY.

Can you post that study up with that inaccuracy range please?

Thanks,

mands

 

[janoshik]

All this info can be provided. I will speak to Jim about it and see if we can link it in the hyperlink on the new thread.

I know this info is all hard data and it would have to be all scanned a posted because it is not in PDF format like the analysis.

Here is what I found going back looking at my notes:

"Amino-acid analysis has a long history in the characterization of protein-based products, since it provides information on the product concentration without referring to an external protein standard and it is independent from the shape and the charge of the protein. In addition, the determined amino-acid composition can confirm sample identity and gives a measure of sample purity. Furthermore, when combined with absorbance measurements, it allows the determination of extinction coefficients under various conditions.1 For protein conjugates, where the synthetic counterpart modifies the protein absorption properties, amino-acid analysis may be required as the only reliable quantification method.

However, in spite of these features, few laboratories can perform such analysis in a reliable and quantitative way, due to the need for specialized equipment and skills. Usually, techniques based on ion-exchange separation coupled with post-column derivatization (e.g., with nin-hydrin, the “classical” method) are considered more precise1 than those based on pre-column derivatization and reversed-phase high-performance liquid chromatography (RP-HPLC), because the latter techniques imply extensive sample manipulation before analysis and are affected by the limited stability of the preformed derivatives.2 However, such RP-HPLC-based methods have the advantage of being accessible to most analytical laboratories, since they do not require expensive dedicated instruments. In addition, manufacturing of dedicated instruments is being halted, making the availability of validated pre-column methods even more important."

mands

Thank you Mr. Mands, I understand it can be a lot of hard work, but I am sure it would help the community. It doesn't hurry at all and I'm all up to interpreting the data and helping how I can.

I totally agree with you that AAA is INVALUABLE at protein characterisation.

But you only have to characterise an UNKNOWN protein. For known proteins characterisation is not necessary at all.

However, when the protein is known, it's far easier and accurate to use the external standard (which is not available with unknown proteins), but readily available with human growth hormone. That's why all pharmaceutical testing methods use it.

As it's written in your notes (which are absolutely correct in everything!!).

Please note, that in your notes it is mentioned that the AAA is independent of protein SHAPE and CHARGE. Shape being ABSOLUTELY necessary for HGH to work. So AAA cannot confirm the shape of the molecule, which actually makes it less useful than most of the HPLC methods.

It is then mentioned that AAA *can* confirm the sample identity, but not that it necessarily does so. Regarding purity AAA is absolutely useless compared to separation methods like HPLC so it has to be used together with them to evaluate purity - which interests much of the community.

Regarding the accuracy of the metod your own notes speak about it.

"few laboratories can perform such analysis in a reliable and quantitative way"

I don't have studies regarding this (and doubt they have been conducted - I can look for them, though), but I have quite some personal experience.

Your notes are mentioning the issue with derivatisation regarding HPLC but his really is not a problem at all with reliable UV detection and standards (which remove the need for derivatisation at all!). So the biggest con of HPLC (in your notes) is simly not a thing anymore in these days.

Also there are many other methods on HPLC, not only RP-HPLC, but SEC-HPLC for example and many others like ion exchange etc.

Can you post that study up with that inaccuracy range please?

Thanks,

mands

See up, Mr. Mands.

It is a pleasure talking to you, Mr. Mands.
You seem to be most reasonable and you have good manners, which I am very thankful for as it makes it easier for me to be as helpful as possible.

I'll be very happy to answer any other questions you might have.


I am not bashing AAA testing at all - au contraire, I'm saying it's invaluable tool.

What I am saying is that Dr. JIM had many statements which were either extremely far stretched or simply untrue regarding the testing method.

 

[Dr JIM]

Do you realise that two separations would be necessary?



.

Two separations is all thats necessary to remove at least 99% of the contaminants from the E-coli supernatant, which is required for Pharmaceutical GH, really!

And you know this but are unaware of how an AAA compares and contrasts with HPLC.

Ok let me get you started, an AAA also involves the use of HPLC. You wanna venture to guess how? This YOU SHOULD KNOW already fella!

 

[Dr JIM]

What I am saying is that Dr. JIM had many statements which were either extremely far stretched or simply untrue regarding the testing method.

POST THEM ALL !

 

[janoshik]

Two separations is all thats necessary to remove at least 99% of the contaminants from the E-coli supernatant, which is required for Pharmaceutical GH, really!

And you know this but are unaware of how an AAA compares and contrasts with HPLC.

Ok let me get you started, an AAA also involves the use of HPLC. You wanna venture to guess how?

You are not too good at reading comprehension, are you?

Two separations are necessary for conduction of AAA analysis.

1st separation is necessary for separating of the protein analysed.

(For members uneducated about the subject - if you mix egg white with HGH and you do AAA analysis straight away, if will give you absolute bullshit for results so you have to separate it first - to get separate egg white and HGH again. if you don't do this, you get bullshit for results - WHICH I BELIEVE MIGHT BE THE CASE OF THIS TESTING! THE RESULTS MIGHT NOT BE AS INACCURATE AS IN CASE OF 'MIXING THE EGG WHITE IN' BUT YOU GET THE POINT).

The 2nd separation is necessary after you isolate the hgh, you subject its 191 amino acids to hydrolysis into separate amino acids. So you have a mixture of amino acids - so how can you find out how many of each amino acid is there? Easiest way is another SEPARATION BY HPLC! So you get each amino acid separate and can 'count it' by absorbance usually.


I know all too well the difference between AAA and HPLC Dr. JIM, you seem to be the uneducated one.

 

[Dr JIM]

Thank you Mr. Mands, I have read it - is it all the raw data available? What about the system used? Separation method used? How it was calibrated?


And no, it is not the most precise method. For quantification it can range up to 15% inaccuracy.

Also please mind that AAA is DEPENDENT on HPLC. HPLC is part of the AAA and AAA can be only as precise as the HPLC separatation used. If there is no separation (how I suspect it was in this case!) beforehands, the results become even less precise.

So in ideal case you have to run 2 HPLCs to get AAA data. So any inaccuracies MULTIPLY.

YES that info IS AVAILABLE but what is your interest in reviewing it, as I'm not willing to jump thru hoops for some UGL detective from Pharmacon (I suspect) until they post their
assays proving what they are producing has been undergone formal analytical testing.

Lets see you cite that info FIRST fella!

 

[PrideofNYC123]

YES that info IS AVAILABLE but what is your interest in reviewing it, as I'm not willing to jump thru hoops for some UGL detective from Pharmacon (I suspect) until they post their
assays proving what they are producing has been undergone formal analytical testing.

Lets see you cite that info FIRST fella!

He's not involved with Pharmacom and isn't the only one interested in the raw data.


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