a few more GENERIC GH ASSAYS

[Dr JIM]

I for one am glad @mands and @Dr JIM are doing what your doing. I know you guys dont have an agenda.

Im grateful meso has you.

Oh I have an agenda but apparently mine is the antithesis of those "do drop in" GH ONLY thread "members".

And the agenda is to uncover the safety and quality/quantity of "generic GH" current being sold on the web and elsewhere, with an emphasis on "higher volume" GH products.

Why would I do such a thing?

There are a number of reasons but a short list includes; the fact the involved labs have declined every opportunity to conduct (and cite) such testing themselves, Meso members AND some of my patients are using this stuff and rightfully have questions that need to be answered from an EVIDENCE BASED perspective.

Jim

 

[ronin17]

Oh I have an agenda but apparently mine is the antithesis of those "do drop in" GH ONLY thread "members".

And the agenda is to uncover the safety and quality/quantity of "generic GH" current being sold on the web and elsewhere, with an emphasis on "higher volume" GH products.

Why would I do such a thing?

There are a number of reasons but a short list includes; the fact the involved labs have declined every opportunity to conduct (and cite) such testing themselves, Meso members AND some of my patients are using this stuff and rightfully have questions that need to be answered from an EVIDENCE BASED perspective.

Jim

That's the kind of agenda I can get behind

 

[Eman]

There are a number of reasons but a short list includes; the fact the involved labs have declined every opportunity to conduct (and cite) such testing themselves, Meso members AND some of my patients are using this stuff and rightfully have questions that need to be answered from an EVIDENCE BASED perspective

Plus you want to know which generic GH brand to use, right Doc?

 

[mands]

Plus you want to know which generic GH brand to use, right Doc?

He's a baby. He can't even hardly run 1-2 iu's without complaining about CTS and such. lol

mands

 

[Eman]

He's a baby. He can't even hardly run 1-2 iu's without complaining about CTS and such. lol

mands

And to think he was only getting .5iu since we've proven pharma can be underdosed and generics are clearly superior.

I can feel Jim's blood pressure going up from here...

 

[ronin17]

And to think he was only getting .5iu since we've proven pharma can be underdosed and generics are clearly superior.

I can feel Jim's blood pressure going up from here...

Fucking priceless. Ha ha.

 

[nolimitz]

Was the serostim from a U.S pharmacy? Is it possible there was some mishandling of how the product was transported and stored which caused it to degrade?

 

[Dr JIM]

Plus you want to know which generic GH brand to use, right Doc?

Because the testing conducted on behalf of MANDS and myself will ever approximate that which is required by Pharma GH manufacturers (except for Zero dosing ) that decision is best deferred to those who elect to use any generic GH product.

And its a risk that is real and should NOT be minimized, in part bc much of it is UNKNOWN especially when GH is used as a PED and results in the development at supra physiologic IGF levels.

However that's not to suggest the data will be posted "in raw form" absent an appropriate practical interpretation to ensure a higher degree of evidence based informed consent.

 

[janoshik]

And IF that is the case EMD is under dosing Serostims bc our ASSAYS are ACCURATE, and consequently matters of this nature will require much more than a tele-promted phone call!

And the FIRST order of business to clarify the issue will be; "where was the product purchased".

I know bc I've already spoken with their "distribution operations manager" who is VERY concerned …….. this fat lady has not sung!

This is probably one of the ways the people can get busted. Anyway, it's very interesting, how you still consider the failure at the side of Serono to be more likely than failure to analyse the sample.

Also, mishandled LYOPHILISED HGH forms aggregates and dimers, if I'm correct.

Those would still get picked as HGH by AAA, due to them being composed from the very same exact amino acids - is it not correct?

However that's not to suggest the data will be posted "in raw form" absent an appropriate practical interpretation to ensure a higher degree of evidence based informed consent.

How convenient. That's exactly something you used to scream and shout censorship about.

Also, with you being the censor, uneducated in the field of analytical chemistry, as it had been proven over and over again, I doubt you can judge what information might be useful and which might not be as necessary.


I don't have any illusions about not being ignored by JIM over and over again, when he lacks counterarguments, but I hope I am helping the members to form their own opinions.

 

[Dr JIM]

Let me put it this way, bc generics have a history of being consistently inconsistent with respect their overall QQ, I would be foolish to accept any level of "risk" that could accompany a manufacturer specific "recommendation".

Oh I can hear it now, yea Jim said Greys are the "best" based on their testing but if that's the case why didn't my IGF levels budge at 10IUs QD and why have I developed this skin rash and elephant like ankles!

JIM

 

[trackstar19]

I've used generics off and on for many years - and AAS on (sadly no off, lol) for 12 years now ... I genuinely appreciate what both Dr. Jim, Mands, ProfessorX, etc. are doing on this board and the testing and getting people to 'think', as well as the testing that the guys over at PM have conducted with Jano, etc.

These tests and conversations only help the community and bring light to things, which everyone should be thankful for. I love science but do not have a science background, so i very much enjoy reading all of this and trying to soak in what I can.

So i know you all argue, etc. but i am sure i speak for many lurkers and otherwise who say THANK YOU.

 

[Brolloks]

Agreed with others that this is a really valuable initiative, and hopefully it won't be sunk by egos.

 

[ProfessorX]

AAA is not a gold standard for pharmaceutical analysis, I've called bullshit on JIM too many times on that already... What they use for routine testing is HPLC methods.



Yeah we get it, you are a big guy.

Jano,

I found this: (not sure if they mean for purified samples for research or if this also refers to formulated biosimilars)

AMINO ACID ANALYSIS - THE GOLD STANDARD FOR PROTEIN QUANTIFICATION

There are several good reasons for why you should use AAA when you need quantitative data

1. It is a very robust analysis. It can be performed in the presence of salt and detergents, as it is based on ion-exchange chromatography. The only contaminant that can obstruct the analysis is a chemical containing primary amines. And even then it might still be possible to do an accurate quantification if the calculations can be based on amino acids not affected by the contaminant.
2. You can compare the results with the sequence. This is a very important feature of the analysis. By comparing the quantitative data for each amino acid residue with the amino acid sequence of the protein you will get information about the purity of your sample. If the data does not fit within a few residues, your sample most likely is contaminated by other proteins.
3. It is linear over a broad concentration range. The analysis can measure from 100-10.000 pmol with high linearity. Therefore, it is suitable for samples with unknown concentration.
4. Your data can be qualified by using internal and external references. The most accurate result is obtained by analyzing each sample in duplicate or triplicate. This means that each sample is analyzed as 2 or 3 individual samples all the way from hydrolysis through data calculation. The precision and the accuracy of the test can be determined using standard proteins in each batch analysis. Adding an internal standard amino acid (e.g. Sarcosine or Norleucine) to each sample makes it possible to correct changes in reagent stability and HPLC flow rate.

(When these new samples are complete......)

I'd like both you and Jim to go over the results of the 4 GH samples that are currently being tested

I should be able to provide detailed analysis information including control standard info (NIST BSA), etc

Hopefully these results will be more accurate than the last samples I had tested using a different lab (same testing method)

In the meantime, I'm getting some info on:

Multiple Reaction Monitoring (MRM)

LC-MS/MS and MRM measurements for quantitative analysis for specific proteins in a complex mixture

Thanks

 

[janoshik]

Jano,

I found this: (not sure if they mean for purified samples for research or if this also refers to formulated biosimilars)

AMINO ACID ANALYSIS - THE GOLD STANDARD FOR PROTEIN QUANTIFICATION

There are several good reasons for why you should use AAA when you need quantitative data

1. It is a very robust analysis. It can be performed in the presence of salt and detergents, as it is based on ion-exchange chromatography. The only contaminant that can obstruct the analysis is a chemical containing primary amines. And even then it might still be possible to do an accurate quantification if the calculations can be based on amino acids not affected by the contaminant.
2. You can compare the results with the sequence. This is a very important feature of the analysis. By comparing the quantitative data for each amino acid residue with the amino acid sequence of the protein you will get information about the purity of your sample. If the data does not fit within a few residues, your sample most likely is contaminated by other proteins.
3. It is linear over a broad concentration range. The analysis can measure from 100-10.000 pmol with high linearity. Therefore, it is suitable for samples with unknown concentration.
4. Your data can be qualified by using internal and external references. The most accurate result is obtained by analyzing each sample in duplicate or triplicate. This means that each sample is analyzed as 2 or 3 individual samples all the way from hydrolysis through data calculation. The precision and the accuracy of the test can be determined using standard proteins in each batch analysis. Adding an internal standard amino acid (e.g. Sarcosine or Norleucine) to each sample makes it possible to correct changes in reagent stability and HPLC flow rate.

(When these new samples are complete......)

I'd like both you and Jim to go over the results of the 4 GH samples that are currently being tested

I should be able to provide detailed analysis information including control standard info (NIST BSA), etc

Hopefully these results will be more accurate than the last samples I had tested using a different lab (same testing method)

In the meantime, I'm getting some info on:

Multiple Reaction Monitoring (MRM)

LC-MS/MS and MRM measurements for quantitative analysis for specific proteins in a complex mixture

Thanks

1. Robustness is a strong point of many other techniques.

2. This is the most awesome thing about AAA. However, if there is reference standard available you don't need to have the sort of information to find out, if the unknown sample is the same as the standard. Also, JIM had talked in his last few posts about quantification from single amino acid - to which I responded that it goes against the biggest benefit of AAA. Just as it can be interpolated from the info you had posted right now.

3. That's linear range of two orders of magnitude. Not uncommon.

4. Again, not uncommon.

Again, as I'm saying AAA is awesome - hell, I'm buying equipment to do it myself soon (not for HGH), but unless coupled with other techniques, it has issues that should not be obfuscated.

AAA unless coupled with other separation techniques is useless as soon as foreign proteins appear in the sample. On the other hand, if with the other separation techniques a standard is used, there's no need for AAA.

I'd be happy if you would post the data, for triplicates of the same sample, for example, so we can assess intraassay deviation.

 

[ProfessorX]

1. Robustness is a strong point of many other techniques.

2. This is the most awesome thing about AAA. However, if there is reference standard available you don't need to have the sort of information to find out, if the unknown sample is the same as the standard. Also, JIM had talked in his last few posts about quantification from single amino acid - to which I responded that it goes against the biggest benefit of AAA. Just as it can be interpolated from the info you had posted right now.

3. That's linear range of two orders of magnitude. Not uncommon.

4. Again, not uncommon.

Again, as I'm saying AAA is awesome - hell, I'm buying equipment to do it myself soon (not for HGH), but unless coupled with other techniques, it has issues that should not be obfuscated.

AAA unless coupled with other separation techniques is useless as soon as foreign proteins appear in the sample. On the other hand, if with the other separation techniques a standard is used, there's no need for AAA.

I'd be happy if you would post the data, for triplicates of the same sample, for example, so we can assess intraassay deviation.

Thank You Jano

As it stands now, the 2 samples I had tested using AAA do not seem to be accurate (lab confirms this with the results from the JTPN sample)

Once the other 4 samples are complete....we will have samples from 3 different labs using the same methodology (AAA) to compare

I do see what Jim is saying about different labs and finding one that is testing properly

I'm still confused by the SEROSTIM result but an easy fix is to just start sending in more "known samples" (PHARMA Samples) to "test" the lab itself

I've done this in the past with SIMEC (HPLC) and my US Toxicology Lab (LC/MS) using tested powders and compound pharmacy finished products, etc

Thanks again for the info

It's appreciated

 

[janoshik]

I'm just happy I can be of help.

Regarding SIMEC, to this day I am not sure what exactly is wrong with their assays of HGH, but at least I think the consensus had been reached that those are wrong.

 

[Dr JIM]

PX reviewing the data from Bio Syn I'm not sure which AA were used to quantify your samples.

Using different AA can create remarkable differences in the results depending upon which AA were readily detectable and reliable enough for quantification purposes.

Bc they should already have the data on hand, let me suggest you ask them to calculate their results using ONLY Alanine, as doing so will enable us to compare SIMILAR assay results.

Finally let me put it this way, after the university lab in LA reaches a similar Sero result as yours and ours
did how much more testing is required
before folk understand the issue is not
a "LAB ERROR" but a sample SOURCE issue.


To that end if you want to "test" a lab use U.S. Pharmaceutical GH ONLY, I've done this with HUMATROPE and Gentropin and have NEVER confronted the problems I'm now seeing "Chinese Pharma products.

Jim

 

[janoshik]

I think that with the method and derivatisation technique described all of them are detectable easily and reliably. Fluorescence is truly awesome thing.

Like it is written in 3 posts above yours, JIM, both by me and in the article quoted by Mr. ProfessorX, using a single amino acid undermines the whole purpose of AAA.

It simply is stupid.

If you are quantifying a single amino acid, you might as well be analysing the vials of HGH by spectrophotometer with built in total protein function.

Anybody can get one for about 1000$. Running costs are like... zero. Work required is like... a minute.

There's a reason it's not done that way and it's not hard to figure it out.


Another post to be conveniently ignored by JIM.

 

[Dr JIM]

Find a lab that stands behind their results PX, and IME the only labs willing to do such a thing are UNIVERSITY based research facilities

.

 

[ProfessorX]

Find a lab that stands behind their results PX, and IME the only labs willing to do such a thing are UNIVERSITY based research facilities

.

I think you mentioned to me your concerns with BIOsy

I think the 2 sample results I received from them prove your concerns to be right

Hopefully the 4 sample results from BioP will be more accurate and comparable to the current lab you are using

I sent you a PM with some additional info