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[159.112.244.3]) by mx.google.com with UTF8SMTPS id af79cd13be357-8d00e60737dsi355527285a.557.2026.03.26.09.51.45 for (version=TLS1_2 cipher=ECDHE-ECDSA-AES128-GCM-SHA256 bits=128/128); Thu, 26 Mar 2026 09:51:45 -0700 (PDT) Received-SPF: pass (google.com: domain of bounce+30b528.39b08c6-tjphuhs=gmail.com@mg-d1.substack.com designates 159.112.244.3 as permitted sender) client-ip=159.112.244.3; Authentication-Results: mx.google.com; dkim=pass header.i=@mg-d1.substack.com header.s=k1 header.b=EvbBwEV8; dkim=pass header.i=@mailgun.org header.s=mg header.b=WkWpGCea; spf=pass (google.com: domain of bounce+30b528.39b08c6-tjphuhs=gmail.com@mg-d1.substack.com designates 159.112.244.3 as permitted sender) smtp.mailfrom="bounce+30b528.39b08c6-tjphuhs=gmail.com@mg-d1.substack.com"; dmarc=pass (p=REJECT sp=REJECT dis=NONE) header.from=substack.com DKIM-Signature: a=rsa-sha256; v=1; c=relaxed/relaxed; d=mg-d1.substack.com; q=dns/txt; s=k1; t=1774543905; x=1774551105; h=List-Unsubscribe-Post: List-Unsubscribe: List-Post: List-Id: List-Archive: List-Owner: Reply-To: In-Reply-To: References: Sender: Sender: Date: Message-Id: To: To: From: From: Subject: Subject: Content-Type: Mime-Version: X-Feedback-Id; bh=G/ClJcaalLrRJwOi+KByS7bfBwSz/uxHX7xvF+xBRvY=; b=EvbBwEV8MJglbuNovM8iMeMdccQbifAVyyK0rm6BNmPYO2XfEKZ+PNUvEB7XgoQKPaC8WSVateb67+JvxgJHfGQz5jhix9U4f1DDqZEmHwRjO88Epil21LmPJ72XgOSZ67n0PtxdVO85tzerE6MS5iTLRkp2ud+s7/HyHEa+U0A= DKIM-Signature: a=rsa-sha256; v=1; c=relaxed/relaxed; d=mailgun.org; q=dns/txt; s=mg; t=1774543905; x=1774551105; h=List-Unsubscribe-Post: List-Unsubscribe: List-Post: List-Id: List-Archive: List-Owner: Reply-To: In-Reply-To: References: Sender: Sender: Date: Message-Id: To: To: From: From: Subject: Subject: Content-Type: Mime-Version: X-Feedback-Id; bh=G/ClJcaalLrRJwOi+KByS7bfBwSz/uxHX7xvF+xBRvY=; b=WkWpGCeaVPruIsZgI2o2AUj436XFhLpGokHKzHSNCdyliH4wFhYoDfebh20oeGqO8fvQQ5aKmjTkaYBiVcXk0n8CzGOTLK9kpokYBuP8aJdzIIbm5tSv1zont7O+x7+wFT6EQ7dNoxVdxMm/rYJlJcKEZvXmCGG4p/E3dX36W0M= X-Mailgun-Sid: WyIxMWU0ZSIsInRqcGh1aHNAZ21haWwuY29tIiwiMzliMDhjNiJd X-Feedback-Id: postmaster@mg-d1.substack.com:post:614a1297c57bbbe25fa40e08:mailgun Received: by 50c736d25c024f216d0e1850e426c186b1b9acfe892fb061219024ef1b763b8e with HTTP id 69c56420f6caf97c2c8cacda; Thu, 26 Mar 2026 16:51:44 GMT X-Mailgun-Sending-Ip: 159.112.244.3 Mime-Version: 1.0 Content-Type: multipart/alternative; boundary="3b2fe90a653273dd6f9a7869024a3de6f84b71e1e1f3c4a2191991cd872d" Subject: Past, Present, and Future: How to Verify a Clean Manufacturing Process From: Derek from Peptide Price To: tjphuhs@gmail.com X-Mailgun-Tag: post X-Mailgun-Track-Clicks: false Message-Id: <20260326165137.3.0dda3d5c6e302ec6@mg-d1.substack.com> Date: Thu, 26 Mar 2026 16:51:37 +0000 Feedback-Id: post-192226265:cat-post:pub-3365367:substack Sender: Derek from Peptide Price References: In-Reply-To: Reply-To: Derek from Peptide Price List-Owner: List-Url: List-Archive: List-Id: List-Post: List-Unsubscribe: List-Unsubscribe-Post: List-Unsubscribe=One-Click X-Mailgun-Variables: {"category": "post", "email_generated_at": "1774543904700", "is_freemail": "true", "post_audience": "everyone", "post_id": "192226265", "post_type": "newsletter", "pub_community_enabled": "true", "publication_id": "3365367", "subdomain": "derekpruski", "user_id": "273622398"} --3b2fe90a653273dd6f9a7869024a3de6f84b71e1e1f3c4a2191991cd872d Content-Type: text/plain; charset="utf-8" Content-Transfer-Encoding: quoted-printable View this post on the web at https://derekpruski.substack.com/p/past-presen= t-and-future-how-to-verify When you look at a certificate of analysis, you=E2=80=99re not looking at o= ne thing. You=E2=80=99re looking at a snapshot across three different windo= ws of time. Each test answers a completely different question =E2=80=94 and= once you understand that, a COA goes from a piece of paper with check mark= s on it to an actual quality story. The Past Test =E2=80=94 USP 85 (Bacterial Endotoxins Test) In plain English: =E2=80=9CWas the manufacturing process clean and controll= ed before it got here?=E2=80=9D USP 85 is the Bacterial Endotoxins Test (BET). It checks for endotoxins =E2= =80=94 toxic remnants left behind by certain gram-negative bacteria during = the manufacturing process. Here=E2=80=99s the key thing most people miss: t= he bacteria themselves don=E2=80=99t have to be alive or present anymore. E= ven dead bacterial cells leave behind lipopolysaccharides (LPS) that can tr= igger serious inflammatory responses in a research subject. A passing USP 85 result tells you the facility that made this compound main= tained controlled, clean manufacturing conditions. It=E2=80=99s a look back= ward =E2=80=94 at the production environment, not what=E2=80=99s in the via= l right now. The Present Test =E2=80=94 HPLC-MS (Purity & Identity) In plain English: =E2=80=9CIs this exactly what it=E2=80=99s supposed to be= , and how pure is it right now?=E2=80=9D High-performance liquid chromatography-mass spectrometry (HPLC-MS) is the g= old standard identity and purity test. It confirms the compound=E2=80=99s m= olecular weight, composition, and sequence, and quantifies exactly how much= of the actual compound is present versus impurities or degradation product= s. This is the test that tells you what=E2=80=99s in the vial today. A compoun= d can come from a clean facility (pass USP 85) and still be mislabeled, mis= dosed, or degraded. HPLC-MS catches all of that. It=E2=80=99s the most crit= ical test on a COA, and it=E2=80=99s the one you should always look for fir= st. The Future Test =E2=80=94 USP 61 (Microbial Enumeration) In plain English: =E2=80=9CIs anything in here that could grow or cause pro= blems over time?=E2=80=9D USP 61 tests for the presence and count of live microbial contamination =E2= =80=94 bacteria, yeast, and mold =E2=80=94 that could proliferate over time= =2E Where USP 85 looks at manufacturing= byproducts, USP 61 looks at living or= ganisms that could be introduced during filling, storage, or handling. This is the forward-looking test. A product can be clean at the time of man= ufacture and still pick up contamination later. USP 61 is the standard micr= obial quality test used broadly across non-sterile pharmaceutical products = and is what most research vendors use. The Sterility Test =E2=80=94 USP 71 In plain English: =E2=80=9CCan we confirm there are absolutely zero viable = microorganisms present?=E2=80=9D USP 71 is the sterility test =E2=80=94 it goes a step further than USP 61 b= y attempting to confirm the complete absence of viable microorganisms. This= is the standard applied to products intended for injectable research and i= s used for injectable research compounds, biologics, and sterile preparatio= ns. The methodology involves incubating samples in growth media over 14 days to= detect even a single colony-forming unit. If something grows, the batch fa= ils. It=E2=80=99s a more rigorous standard, which is exactly why it carries= different regulatory implications =E2=80=94 and why you don=E2=80=99t see = it universally across research vendors. Quick Reference USP 85 =E2=80=94 Past =E2=86=92 Was manufacturing clean? HPLC-MS =E2=80=94 Present =E2=86=92 Is it correct and pure right now? USP 61 =E2=80=94 Future =E2=86=92 Will anything grow over time? USP 71 =E2=80=94 Future+ =E2=86=92 Confirmed sterility (injectable research= standard) Why Some Companies Opt for USP 61 Instead of USP 71 =E2=80=94 Something to = Keep in Mind USP 61 and USP 71 are often treated as interchangeable in the research spac= e =E2=80=94 and from a pure microbiology standpoint, they=E2=80=99re measur= ing similar things. Both look for live microbial contamination. But there= =E2=80=99s a reason some companies deliberately choose USP 61 over USP 71, = and it has nothing to do with cutting corners. USP 71 is the sterility standard for injectable research. Performing USP 71= testing signals =E2=80=94 or can be interpreted as signaling =E2=80=94 tha= t a product is intended for injectable research. In a space where everythin= g is labeled research use only, not for human consumption, that distinction= carries real legal weight. I=E2=80=99ve spoken directly with research comp= anies whose legal teams have explicitly prohibited them from conducting USP= 71 testing for exactly this reason =E2=80=94 not because the test itself i= s problematic, but because it can be used to imply an injectable research c= lassification that exposes the business to regulatory risk. It=E2=80=99s a fine line, and the research space is still very undefined. E= very company is going to have a different risk tolerance and operate accord= ingly. Some will opt for USP 71 and accept that tradeoff. Others will stick= with USP 61 on the advice of legal counsel. Neither position is inherently= wrong =E2=80=94 it=E2=80=99s a business decision made in a space where the= rules aren=E2=80=99t fully written yet. What=E2=80=99s worth knowing: the cost difference between the two tests is = essentially zero. So when you see USP 61 on a COA, it=E2=80=99s not a budge= t decision. It=E2=80=99s likely a legal one. Keep this in the back of your mind when evaluating COAs. The presence of an= y microbial testing is a positive signal =E2=80=94 and the gap between a ve= ndor that does microbial testing and one that doesn=E2=80=99t is far more m= eaningful than whether they chose 61 or 71. Next time you pull up a COA, run through the three windows mentally. USP 85= =E2=80=94 was it made cleanly? HPLC-MS =E2=80=94 is it what it says it is = right now? USP 61 or 71 =E2=80=94 is anything alive in there? When a vendor= hits all three, that=E2=80=99s a genuinely comprehensive quality picture. = Whether they chose 61 or 71 often comes down to how they approach the legal= side of things =E2=80=94 both signal that they care about sterility and ba= cterial growth. When a vendor is missing one of the three windows entirely,= you now know exactly what question isn=E2=80=99t being answered. Unsubscribe https://substack.com/redirect/2/eyJlIjoiaHR0cHM6Ly9kZXJla3BydXN= raS5zdWJzdGFjay5jb20vYWN0aW9uL2Rpc2FibGVfZW1haWw_dG9rZW49ZXlKMWMyVnlYMmxrSW= pveU56TTJNakl6T1Rnc0luQnZjM1JmYVdRaU9qRTVNakl5TmpJMk5Td2lhV0YwSWpveE56YzBOV= FF6T1RBMExDSmxlSEFpT2pFNE1EWXdOems1TURRc0ltbHpjeUk2SW5CMVlpMHpNelkxTXpZM0lp= d2ljM1ZpSWpvaVpHbHpZV0pzWlY5bGJXRnBiQ0o5Lmc0ZlA3VFZjaTdtRjhqNTZWaEFCSmJKVnp= CUTY5b1NsM2tUeWxPWlQyRkEiLCJwIjoxOTIyMjYyNjUsInMiOjMzNjUzNjcsImYiOnRydWUsIn= UiOjI3MzYyMjM5OCwiaWF0IjoxNzc0NTQzOTA0LCJleHAiOjIwOTAxMTk5MDQsImlzcyI6InB1Y= i0wIiwic3ViIjoibGluay1yZWRpcmVjdCJ9.3i3AStpwCH2TV0BIP0PC1PyZkjp0ueaA5KcwrRj= DuHo? --3b2fe90a653273dd6f9a7869024a3de6f84b71e1e1f3c4a2191991cd872d Content-Type: text/html; charset="utf-8" Content-Transfer-Encoding: quoted-printable Past, Present, and Futur= e: How to Verify a Clean Manufacturing Process3D""
When you look at a certificate of analysis, you’re not lookin= g at one thing.
͏     ­͏     ­&#= 847;     ­͏     ­͏    = ­͏     ­͏     ­͏  = ;   ­͏     ­͏     ­= 47;     ­͏     ­͏     = ­͏     ­͏     ­͏  =   ­͏     ­͏     ­T= 7;     ­͏     ­͏     &= #173;͏     ­͏     ­͏   =   ­͏     ­͏     ­͏= ;     ­͏     ­͏     &#= 173;͏     ­͏     ­͏   &= #8199; ­͏     ­͏     ­͏=     ­͏     ­͏     = 73;͏     ­͏     ­͏   &#= 8199; ­͏     ­͏     ­͏ =     ­͏     ­͏     = 3;͏     ­͏     ­͏   = 199; ­͏     ­͏     ­͏ &= nbsp;   ­͏     ­͏     ­= ;͏     ­͏     ­͏   Q= 99; ­͏     ­͏     ­͏ &n= bsp;   ­͏     ­͏     ­= ͏     ­͏     ­͏   ̳= 9; ­͏     ­͏     ­͏ &nb= sp;   ­͏     ­͏     ­&= #847;     ­͏     ­͏    = ; ­͏     ­͏     ­͏ &nbs= p;   ­͏     ­͏     ­&#= 847;     ­͏     ­͏    = ­͏     ­͏     ­͏  = ;   ­͏     ­͏     ­= 47;     ­͏     ­͏     = ­͏     ­͏     ­͏  =   ­͏     ­͏     ­T= 7;     ­͏     ­͏     &= #173;͏     ­͏     ­͏   =   ­͏     ­͏     ­͏= ;     ­͏     ­͏     &#= 173;͏     ­͏     ­͏   &= #8199; ­͏     ­͏     ­͏=     ­͏     ­͏     = 73;͏     ­͏     ­͏   &#= 8199; ­͏     ­͏     ­͏ =     ­͏     ­͏     = 3;͏     ­͏     ­͏   = 199; ­͏     ­͏     ­͏ &= nbsp;   ­͏     ­͏     ­= ;͏     ­͏     ­͏   Q= 99; ­͏     ­͏     ­͏ &n= bsp;   ­͏     ­͏     ­= ͏     ­͏     ­͏   ̳= 9; ­͏     ­͏     ­͏ &nb= sp;   ­͏     ­͏     ­&= #847;     ­͏     ­͏    = ; ­͏     ­͏     ­͏ &nbs= p;   ­͏     ­͏     ­&#= 847;     ­͏     ­͏    = ­͏     ­͏     ­͏  = ;   ­͏     ­͏     ­= 47;     ­͏     ­͏     = ­͏     ­͏     ­͏  =   ­͏     ­͏     ­T= 7;     ­͏     ­͏     &= #173;͏     ­͏     ­͏   =   ­͏     ­͏     ­͏= ;     ­͏     ­͏     &#= 173;͏     ­͏     ­͏   &= #8199; ­͏     ­͏     ­͏=     ­͏     ­͏     = 73;͏     ­͏     ­͏   &#= 8199; ­͏     ­͏     ­͏ =     ­͏     ­͏     = 3;͏     ­͏     ­͏   = 199; ­͏     ­͏     ­
=
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Past, Present, an= d Future: How to Verify a Clean Manufacturing Process

Derek
 = ;
3D""
= 3D""
3D""
3D""
READ = IN APP3D""
=
 

When you look at a certificate of analysis, you’re not looking= at one thing. You’re looking at a snapshot across three different wi= ndows of time. Each test answers a completely different question — an= d once you understand that, a COA goes from a piece of paper with check mar= ks on it to an actual quality story.

<= tbody>
3D""
<= /figure>

The Past Test — USP 85 (B= acterial Endotoxins Test)

In plain English: “= ;Was the manufacturing process clean and controlled before it got here?R= 21;

USP 85 is the Bacterial Endotoxins Test (BET).= It checks for endotoxins — toxic remnants left behind by certain gra= m-negative bacteria during the manufacturing process. Here’s the key = thing most people miss: the bacteria themselves don’t have to be aliv= e or present anymore. Even dead bacterial cells leave behind lipopolysaccha= rides (LPS) that can trigger serious inflammatory responses in a research s= ubject.

A passing USP 85 result tells you the facility that= made this compound maintained controlled, clean manufacturing conditions. = It’s a look backward — at the production environment, not what&= #8217;s in the vial right now.

The Present Test — HPL= C-MS (Purity & Identity)

In plain English: = 220;Is this exactly what it’s supposed to be, and how pure is it righ= t now?”

High-performance liquid chromatograp= hy-mass spectrometry (HPLC-MS) is the gold standard identity and purity tes= t. It confirms the compound’s molecular weight, composition, and sequ= ence, and quantifies exactly how much of the actual compound is present ver= sus impurities or degradation products.

This is the test th= at tells you what’s in the vial today. A compound can come from a cle= an facility (pass USP 85) and still be mislabeled, misdosed, or degraded. H= PLC-MS catches all of that. It’s the most critical test on a COA, and= it’s the one you should always look for first.

The F= uture Test — USP 61 (Microbial Enumeration)

= In plain English: “Is anything in here that could grow or cause probl= ems over time?”

USP 61 tests for the presenc= e and count of live microbial contamination — bacteria, yeast, and mo= ld — that could proliferate over time. Where USP 85 looks at manufact= uring byproducts, USP 61 looks at living organisms that could be introduced= during filling, storage, or handling.

This is the forward-= looking test. A product can be clean at the time of manufacture and still p= ick up contamination later. USP 61 is the standard microbial quality test u= sed broadly across non-sterile pharmaceutical products and is what most res= earch vendors use.

The Sterility Test — USP 71

In plain English: “Can we confirm there are absolu= tely zero viable microorganisms present?”

US= P 71 is the sterility test — it goes a step further than USP 61 by at= tempting to confirm the complete absence of viable microorganisms. This is = the standard applied to products intended for injectable research and is us= ed for injectable research compounds, biologics, and sterile preparations.<= /p>

The methodology involves incubating samples in growth media= over 14 days to detect even a single colony-forming unit. If something gro= ws, the batch fails. It’s a more rigorous standard, which is exactly = why it carries different regulatory implications — and why you don= 217;t see it universally across research vendors.

Quick Ref= erence

  • USP 85 — Past →= ; Was manufacturing clean?

  • HPLC-MS — Present → Is = it correct and pure right now?

  • USP 61 — Future → W= ill anything grow over time?

  • USP 71 — Future+ → Co= nfirmed sterility (injectable research standard)

Why Some Companies Opt for USP 61 Instead of USP 71 — Somethin= g to Keep in Mind

USP 61 and USP 71 are often treated as i= nterchangeable in the research space — and from a pure microbiology s= tandpoint, they’re measuring similar things. Both look for live micro= bial contamination. But there’s a reason some companies deliberately = choose USP 61 over USP 71, and it has nothing to do with cutting corners.

USP 71 is the sterility standard for injectable research. Pe= rforming USP 71 testing signals — or can be interpreted as signaling = — that a product is intended for injectable research. In a space wher= e everything is labeled research use only, not for human consumption, that = distinction carries real legal weight. I’ve spoken directly with rese= arch companies whose legal teams have explicitly prohibited them from condu= cting USP 71 testing for exactly this reason — not because the test i= tself is problematic, but because it can be used to imply an injectable res= earch classification that exposes the business to regulatory risk.

It’s a fine line, and the research space is still very undefi= ned. Every company is going to have a different risk tolerance and operate = accordingly. Some will opt for USP 71 and accept that tradeoff. Others will= stick with USP 61 on the advice of legal counsel. Neither position is inhe= rently wrong — it’s a business decision made in a space where t= he rules aren’t fully written yet.

What’s worth= knowing: the cost difference between the two tests is essentially zero. So= when you see USP 61 on a COA, it’s not a budget decision. It’s= likely a legal one.

Keep this in the back of your mind whe= n evaluating COAs. The presence of any microbial testing is a positive sign= al — and the gap between a vendor that does microbial testing and one= that doesn’t is far more meaningful than whether they chose 61 or 71= =2E

Next time you pull up a COA, run through the three windows = mentally. USP 85 — was it made cleanly? HPLC-MS — is it what it= says it is right now? USP 61 or 71 — is anything alive in there? Whe= n a vendor hits all three, that’s a genuinely comprehensive quality p= icture. Whether they chose 61 or 71 often comes down to how they approach t= he legal side of things — both signal that they care about sterility = and bacterial growth. When a vendor is missing one of the three windows ent= irely, you now know exactly what question isn’t being answered.

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