Quality Control & Certificates of Analysis

Quick Navigation

• Understanding Purity
• Target vs Overall Purity
• Analytical Methods
• COA Components
• Authentic vs Fake COAs
• Red Flags
• Verification Checklist
• Example Analysis

Understanding Peptide Purity

Peptide purity is a critical quality parameter that indicates the percentage of the desired peptide in a sample relative to impurities. However, "purity" is not a single, simple measurement - it encompasses several distinct concepts that are often confused or misrepresented.

Target Purity vs Overall Purity

The Three Pillars of Peptide Purity

Analytical Methods for Peptide Characterization

1. High-Performance Liquid Chromatography (HPLC)

2. Mass Spectrometry (MS/MALDI-TOF)

3. Amino Acid Analysis (AAA)

4. Karl Fischer Titration

Certificate of Analysis (COA) Components

A comprehensive Certificate of Analysis should contain the following essential information:

1. Product Identification

• Peptide name (full name and abbreviation)
• Catalog/Lot number
• Sequence (single-letter or three-letter code)
• Molecular formula and theoretical molecular weight
• CAS number (if applicable)

2. Supplier Information

• Company name and full address
• Contact information (phone, email)
• Manufacturing/testing date
• Expiration/retest date

3. Analytical Data

• HPLC purity: Chromatogram showing peak area percentage
• Mass spectrometry: Observed mass vs calculated mass
• Peptide content (AAA): Percentage by weight (critical but often omitted)
• Counter-ion: Type (TFA, acetate, HCl) and percentage
• Water content: Percentage (Karl Fischer)
• Appearance: Physical description (white powder, etc.)

4. Chromatogram Requirements (HPLC)

• X-axis: Retention time (minutes)
• Y-axis: Absorbance (mAU) at specified wavelength
• Clear baseline and peak resolution
• Main peak clearly labeled with retention time and area %
• Column specifications (type, dimensions, particle size)
• Mobile phase composition and gradient program
• Flow rate and injection volume
• Temperature

5. Mass Spectrum Requirements

• Observed m/z peaks
• Calculated theoretical mass
• Mass accuracy (Δ ppm)
• Ionization mode (ESI, MALDI)
• Adduct ions identified ([M+H]+, [M+2H]2+, [M+Na]+)

6. Quality Assurance

• Analyst name/initials and signature
• QC approval signature
• Laboratory accreditation information (if applicable)
• Reference to analytical methods/SOPs

Authentic vs Fraudulent Certificates of Analysis

The peptide market is rife with counterfeit products and fraudulent COAs. Understanding the differences between authentic and fake documentation is essential for ensuring product quality and safety.

Common Fraudulent Practices

COA Verification Checklist

Use this checklist to evaluate any Certificate of Analysis:

Level 1: Critical Must-Haves (Non-negotiable)

• Complete supplier contact information (name, address, phone)
• Specific lot/batch number matching your product
• HPLC chromatogram (full graph, not just percentage)
• Mass spectrometry data confirming molecular weight
• Testing date within reasonable timeframe

Level 2: Important Quality Indicators

• Peptide content by AAA or equivalent method
• Counter-ion type and percentage
• Water content (Karl Fischer)
• Detailed HPLC method parameters
• Signed by identifiable personnel
• Impurity information if applicable

Level 3: Professional Standards

• Multiple orthogonal analytical methods
• Reference to internal SOPs or USP methods
• Clear distinction between target and overall purity
• Accreditation information (ISO 17025, GMP, etc.)
• Appropriate language and technical terminology
• High-quality formatting and presentation

Immediate Red Flags (Walk Away)

• No batch number or doesn't match product label
• Only purity percentage - no raw analytical data
• Seller refuses to provide COA or only provides after payment
• COA is clearly a screenshot or low-quality scan
• Claims of "99.9%" or "100%" purity
• No mass spectrometry data
• Anonymous or generic sender ("Lab Director", "QC Team")
• Spelling errors, poor grammar, unprofessional format
• Missing company name or unverifiable address
• COA date is years old

Example COA Analysis

Example 1: Authentic COA (BPC-157)

═══════════════════════════════════════════════════════════════
                CERTIFICATE OF ANALYSIS
═══════════════════════════════════════════════════════════════

Peptide Sciences Inc.
123 Research Park Drive
Biotechnology Plaza, Suite 400
San Diego, CA 92121
Phone: +1-858-555-0100 | Email: qc@peptidesciences.com

Product Name:      BPC-157 (Body Protection Compound 157)
Catalog Number:    PS-BPC-001
Lot Number:        BPC157-2024-Q2-0847
Manufacturing Date: April 15, 2024
Expiration Date:   April 15, 2026
Storage:           -20°C, protect from light

───────────────────────────────────────────────────────────────
PEPTIDE INFORMATION
───────────────────────────────────────────────────────────────
Sequence:          Gly-Glu-Pro-Pro-Pro-Gly-Lys-Pro-Ala-Asp-Asp-Ala-Gly-Leu-Val
                   (GEPPP GKPAD DAGLV)
Molecular Formula: C62H98N16O22
Molecular Weight:  1419.53 Da (calculated)
Counter-ion:       Acetate salt

───────────────────────────────────────────────────────────────
QUALITY CONTROL DATA
───────────────────────────────────────────────────────────────

1. TARGET PURITY (HPLC)
   Method:         Reverse-Phase HPLC
   Column:         Phenomenex Luna C18(2), 250 x 4.6 mm, 5 μm
   Mobile Phase A: 0.1% TFA in water
   Mobile Phase B: 0.1% TFA in acetonitrile
   Gradient:       5-95% B over 30 min
   Flow Rate:      1.0 mL/min
   Detection:      UV 220 nm
   Injection:      20 μL
   Temperature:    25°C

   Result: 96.8% (Area under curve)
   Main Peak RT:   18.45 min (96.8%)
   Impurities:     RT 17.2 min (1.4%)
                   RT 19.8 min (1.2%)
                   Other peaks: <0.6%

2. IDENTITY CONFIRMATION (Mass Spectrometry)
   Method:         ESI-MS (Electrospray Ionization)
   Instrument:     Agilent 6545 Q-TOF

   Calculated Mass: 1419.53 Da
   Observed Mass:  1419.48 Da
   Δ Mass:         -0.05 Da (-35 ppm)

   Major Ions Detected:
   [M+H]+:     m/z 1420.50
   [M+2H]2+:   m/z 710.75
   [M+3H]3+:   m/z 474.17

   ✓ Molecular identity CONFIRMED

3. PEPTIDE CONTENT (Amino Acid Analysis)
   Method:         AAA after 6M HCl hydrolysis (24h, 110°C)

   Expected Ratio  Observed Ratio  %Recovery
   Gly:    3.0     2.98           99.3%
   Pro:    4.0     3.92           98.0%
   Glu:    1.0     0.97           97.0%
   Lys:    1.0     1.02           102.0%
   Ala:    2.0     1.95           97.5%
   Asp:    2.0     2.04           102.0%
   Leu:    1.0     0.99           99.0%
   Val:    1.0     0.98           98.0%

   Total Peptide Content: 78.3% (w/w)

   Breakdown of Sample Composition:
   - BPC-157 peptide:    78.3%
   - Acetate counter-ion: 12.1%
   - Water (Karl Fischer): 8.4%
   - Other:               1.2%

4. PHYSICAL PROPERTIES
   Appearance:     White to off-white lyophilized powder
   Solubility:     Soluble in water, saline, or sterile PBS
   pH (1% solution): 4.5-5.5

───────────────────────────────────────────────────────────────
SPECIFICATIONS
───────────────────────────────────────────────────────────────
Target Purity (HPLC):    ≥ 95%           PASS (96.8%)
Peptide Content (AAA):   ≥ 75%           PASS (78.3%)
Mass Accuracy:           ±0.1%           PASS (-0.05 Da)
Water Content:           ≤ 10%           PASS (8.4%)

═══════════════════════════════════════════════════════════════

Analyzed by:    Dr. Sarah Chen, PhD        Date: April 18, 2024
                Senior Analytical Chemist

Approved by:    Dr. Michael Torres, PhD    Date: April 19, 2024
                Director of Quality Control

This Certificate of Analysis is issued in accordance with ISO
17025:2017 standards. Peptide Sciences is GMP compliant.

═══════════════════════════════════════════════════════════════

Analysis of Authentic COA:

• Complete contact information: Full company name, address, phone, email
• Specific lot number: BPC157-2024-Q2-0847 (unique and traceable)
• Comprehensive data: HPLC + MS + AAA (three orthogonal methods)
• Realistic values: 96.8% HPLC purity (not "99.9%")
• Both purities reported: Target purity (96.8%) AND peptide content (78.3%)
• Detailed methods: Complete HPLC parameters, column specs, gradient program
• Impurity disclosure: Shows minor impurity peaks (realistic)
• Mass accuracy: Within acceptable range (±35 ppm)
• Counter-ion specified: Acetate salt with percentage
• Water content: Measured by Karl Fischer (8.4%)
• Named personnel: Signed by specific individuals with credentials
• Professional presentation: Well-formatted, technical terminology
• Quality standards: References ISO 17025 and GMP compliance

Conclusion: This COA provides comprehensive, verifiable data and meets professional standards for peptide quality documentation.

Example 2: Fraudulent COA (Red Flags)

═══════════════════════════════════════════════════════════════
                  CERTIFICATE OF ANALYSIS
═══════════════════════════════════════════════════════════════

Global Peptides Ltd
China                           ← ⚠️ No specific address

Product:     BPC-157
Batch:       00123             ← ⚠️ Generic, non-specific lot number
Date:        2023-01-05        ← ⚠️ Over 1 year old

───────────────────────────────────────────────────────────────
ANALYSIS RESULTS
───────────────────────────────────────────────────────────────

Purity: 99.9%        ← ⚠️ Suspiciously perfect purity
                          ⚠️ No method specified (HPLC? MS?)
                          ⚠️ No raw data provided

Molecular Weight: Confirmed  ← ⚠️ No actual mass values
                                  ⚠️ No mass spectrum provided

Appearance: White Powder

Quality: High Qualty Product  ← ⚠️ Spelling error ("Qualty")
                                    ⚠️ Vague, non-technical language

───────────────────────────────────────────────────────────────

Tested by: QC Department   ← ⚠️ Anonymous, no named personnel
Approved: Manager           ← ⚠️ No credentials or qualifications

═══════════════════════════════════════════════════════════════

Critical Failures in Fraudulent COA:

• No complete address: Just "China" - untraceable
• Generic batch number: "00123" - not unique or informative
• Old date: Over 1 year old - questionable relevance
• No analytical methods: Doesn't specify HPLC, MS, or any technique
• Unrealistic purity: "99.9%" is suspiciously perfect
• No raw data: No chromatogram, no mass spectrum
• Vague mass confirmation: No actual observed/calculated masses
• No peptide content: Only "purity" - likely misleading
• Spelling error: "High Qualty" indicates unprofessional documentation
• Anonymous signatories: "QC Department" and "Manager" with no names
• No methodology details: Missing all technical parameters
• No impurity data: Unrealistic to have exactly 99.9% purity

Best Practices for Consumers

1. Before Purchase

• Request COA before ordering
• Verify supplier credentials and reputation
• Check for third-party testing or accreditation
• Compare prices - suspiciously low prices indicate lower quality
• Research supplier reviews on independent forums

2. Upon Receipt

• Verify lot number on vial matches COA
• Check physical appearance matches COA description
• Store immediately per COA recommendations
• Consider third-party testing for expensive or critical peptides

3. For Dosing

• Use peptide content (AAA), not HPLC purity, for dose calculations
• Account for water content when weighing
• Factor in counter-ion mass
• If peptide content not provided, assume 70-75% for acetate salts, 60-65% for TFA salts

4. Red Flags to Avoid

• Sellers unwilling to provide COA before purchase
• COAs that are clearly generic or recycled
• Prices significantly below market average
• Lack of contact information or verifiable address
• Pressure tactics or "limited time" sales
• Claims of pharmaceutical-grade without evidence

Educational Disclaimer