1. Analytical Method & Instrumentation
Method: Reversed-Phase HPLC per USP <621>
Instrument: Agilent 1260 Infinity II HPLC System (SN: DE75555)
Column: Agilent ZORBAX Eclipse Plus C18, 250 x 4.6 mm, 5 μm (Lot: B92718)
Mobile Phase A: 0.1% Trifluoroacetic acid (TFA) in Milli-Q Water (18.2 MΩ·cm)
Mobile Phase B: 0.1% TFA in Acetonitrile (HPLC Grade, Fisher Scientific)
Gradient Program: 0 min → 5%B, 5 min → 20%B, 20 min → 60%B, 28 min → 95%B, 30 min → 95%B
Flow Rate / Temp: 1.0 mL/min / 30.0 ± 0.5°C (thermostatted column compartment)
Detection: UV-DAD at 220 nm (bandwidth 4 nm, ref. 360 nm)
Injection: 20 μL, sample conc. 0.5 mg/mL in 0.1% TFA/H₂O
Data Processing: Agilent OpenLAB CDS ChemStation Rev. C.01.10
2. System Suitability Test (SST)
Parameter Acceptance Criteria Result Status
Theoretical Plates (N) ≥ 2000 15363 PASS Tailing Factor (T) 0.8 – 1.5 1.07 PASS Repeatability RSD (n=6) ≤ 1.0% 0.24% PASS Resolution (Rs) ≥ 1.5 2.8 PASS
3. Purity & Assay Results
Test Item Specification Result Status
Appearance White to off-white lyophilized powder Conforms PASS Identification (FTIR) Consistent with reference spectrum Consistent PASS Purity (RP-HPLC, area%) ≥98% 98.2% PASS Assay (Potentiometric Titration) 98.0% – 102.0% 99.9% PASS Moisture (Karl Fischer, V-30S) ≤ 1.0% 0.45% PASS Largest Single Impurity ≤ 0.50% 0.21% PASS Total Impurities ≤ 2.0% 1.80% PASS Specific Rotation [α]²⁰D Literature value ± 2.0° +5.9° (c=1, DMF) PASS Residual Solvents (GC-FID/HS, per ICH Q3C R8) DMF (Class 2) ≤ 880 ppm 203 ppm PASS DCM (Class 2) ≤ 600 ppm 94 ppm PASS Methanol (Class 2) ≤ 3000 ppm 236 ppm PASS Acetonitrile (Class 2) ≤ 410 ppm 32 ppm PASS Ethyl Acetate (Class 3) ≤ 5000 ppm 105 ppm PASS
4. HPLC Chromatogram
Fig. 1: RP-HPLC chromatogram of Winstrol Water (Stanozolol) (Batch: COA-SW50-3679). Main peak RT = — min, Area% = 98.2%. UV detection at 220 nm.
1. Analytical Method & Instrumentation
Standard: ICP-MS per ICH Q3D(R2) / USP <232> <233>
Instrument: Agilent 7900 ICP-MS (SN: JP34714) with He collision mode
Sample Preparation: Closed-vessel microwave digestion (CEM MARS 6): 48.8 mg + 5 mL HNO₃ (suprapure) + 1 mL H₂O₂ (30%)
Digestion Program: Ramp 20 min → 200°C, hold 15 min, cool to RT, dilute to 50.0 mL (Milli-Q)
Calibration: Multi-element std (NIST SRM 3100 series), 5-point external cal. (r² ≥ 0.999)
Internal Standards: ⁴⁵Sc, ⁷²Ge, ¹⁰³Rh, ¹⁸⁵Re, ²⁰⁹Bi — 10 ppb each, online addition
Quality Control: Method blank, CRM (NIST 1577c), spike recovery (90–110%), duplicate analysis
2. Elemental Impurity Results
Element ICH Class PDE Oral(μg/day) Limit(ppm) Result(ppm) Status
Lead (Pb) — m/z 208 1 5 ≤ 0.5 0.021 PASS Cadmium (Cd) — m/z 111 1 5 ≤ 0.5 0.054 PASS Arsenic (As) — m/z 75 1 15 ≤ 1.5 0.257 PASS Mercury (Hg) — m/z 202 1 30 ≤ 3.0 0.023 PASS Cobalt (Co) — m/z 59 2A 50 ≤ 5.0 0.126 PASS Vanadium (V) — m/z 51 2A 100 ≤ 10 0.023 PASS Nickel (Ni) — m/z 60 2A 200 ≤ 20 1.345 PASS Palladium (Pd) — m/z 105 2B 100 ≤ 10 0.195 PASS Platinum (Pt) — m/z 195 2B 100 ≤ 10 < LOD PASS Silver (Ag) — m/z 107 2B 150 ≤ 15 0.127 PASS Copper (Cu) — m/z 63 3 3000 ≤ 300 1.33 PASS Iron (Fe) — m/z 56 3 — ≤ 100 3.86 PASS Nickel (Ni) — m/z 60 (low-tox) 3 — ≤ 200 0.24 PASS Zinc (Zn) — m/z 66 3 13000 ≤ 1300 0.75 PASS Chromium (Cr) — m/z 52 3 11000 ≤ 1100 0.463 PASS Manganese (Mn) — m/z 55 3 2500 ≤ 250 0.95 PASS
3. Method Validation Parameters
Parameter Criteria Result Status
LOD (Class 1 Elements) ≤ J/10 of limit As: 0.001, Cd: 0.0005, Hg: 0.0003, Pb: 0.001 ppm PASS LOQ (Class 1 Elements) ≤ J/2 of limit As: 0.005, Cd: 0.002, Hg: 0.001, Pb: 0.005 ppm PASS Spike Recovery 70–150% (per USP <233>) 100.8% – 103.4% PASS Repeatability (n=3) RSD ≤ 20% 6.3% PASS Method Blank < LOQ for all elements All < LOQ PASS CRM Recovery (NIST 1577c) 80–120% certified values 102.1% PASS
1. Test Method & Conditions
Standard: USP <71> Sterility Tests / EP 2.6.1
Method: Membrane Filtration (0.45 μm cellulose nitrate, Millipore HAWP)
Rinse Solution: 0.1% Peptone Water (Fluid A per USP), 3 × 100 mL per membrane
Sample Size: 48.8 mg dissolved in 10 mL sterile diluent, filtered through 2 membranes
Media: FTM (Fluid Thioglycollate Medium) + SCDM (Soybean-Casein Digest Medium), BD Difco
Environment: ISO Class 5 (Grade A) / ISO Class 7 background, ΔP ≥ 15 Pa
Incubators: Validated Memmert IN110 ± 0.5°C, IQ/OQ/PQ qualified, continuous temperature logging
2. Growth Promotion Test (Media Sensitivity Verification)
Challenge Organism ATCC No. Medium Temp. Inoculum Growth Status
Staphylococcus aureus ATCC 6538 FTM 32.5°C < 100 CFU Turbid < 72h PASS Bacillus subtilis ATCC 6633 FTM 32.5°C < 100 CFU Turbid < 72h PASS Clostridium sporogenes ATCC 19404 FTM 32.5°C < 100 CFU Turbid < 72h PASS Candida albicans ATCC 10231 SCDM 22.5°C < 100 CFU Turbid < 5d PASS Aspergillus brasiliensis ATCC 16404 SCDM 22.5°C < 100 CFU Turbid < 5d PASS
3. Sterility Test Results
Test Medium Temp. Period Day 7 Day 14 Status
Aerobic Bacteria FTM (2 tubes) 30–35°C 14 days Clear Clear PASS Anaerobic Bacteria FTM (2 tubes) 30–35°C 14 days Clear Clear PASS Fungi SCDM (2 tubes) 20–25°C 14 days Clear Clear PASS Yeasts SCDM (2 tubes) 20–25°C 14 days Clear Clear PASS Negative Control FTM + SCDM As above 14 days Clear Clear PASS
4. Microbial Enumeration (USP <61> / <62>)
Test Item Method Limit Result Status
Total Aerobic Microbial Count (TAMC) Pour plate / TSA / 32.5°C / 5d ≤ 10² CFU/g < 10 CFU/g PASS Total Yeast & Mold Count (TYMC) Pour plate / SDA / 22.5°C / 5d ≤ 10¹ CFU/g < 10 CFU/g PASS Escherichia coli Enrichment + MacConkey Absent / 1 g Not Detected PASS Salmonella spp.Enrichment + XLD/HE agar Absent / 10 g Not Detected PASS Staphylococcus aureus Enrichment + Baird-Parker Absent / 1 g Not Detected PASS Pseudomonas aeruginosa Enrichment + Cetrimide Absent / 1 g Not Detected PASS Bile-Tolerant Gram-Negative (BTGNB) Enrichment + VRBGA ≤ 10² CFU/g Not Detected PASS
1. Test Method & Instrumentation
Standard: USP <85> Bacterial Endotoxins Test / EP 2.6.14
Method: Kinetic Turbidimetric LAL Assay
Instrument: Lonza PyroGene™ rFC Detection System + BioTek ELx808 microplate reader
LAL Reagent: Lonza PYROGENT™ Plus, λ = 0.03 EU/mL (Lot: L7872, Exp: 2026-09)
Reference Std. (RSE): USP Reference Standard Endotoxin (E. coli O55:B5), Potency: 10,000 EU/vial
LAL Reagent Water: Lonza LAL Water, tested < 0.005 EU/mL (Lot: W9532)
Sample Preparation: 48.8 mg in 1.0 mL LAL water → vortex 30s → serial dilution to MVD
2. Endotoxin Limit Calculation
Parameter Value
Maximum Daily Dose (M) 10 mg/kg (assumed max parenteral) Threshold Pyrogenic Dose (K) 5.0 EU/kg/hr (parenteral, non-intrathecal) Endotoxin Limit = K/M 0.50 EU/mg Maximum Valid Dilution (MVD) 1:64 Actual Test Concentration 0.15 mg/mL (diluted to MVD)
3. System Suitability Validation
Validation Item Acceptance Criteria Result Status
Standard Curve |r| Correlation |r| ≥ 0.980 0.9979 PASS Standard Curve Slope −1.0 ± 0.5 (log-log) −1.103 PASS Standard CV (each conc.) ≤ 25% Max: 13.0% PASS Positive Product Control (PPC) Recovery 50–200% 111.5% PASS Negative Control < λ (0.03 EU/mL) < 0.005 EU/mL PASS Interference Test Recovery 50–200% 96.5% PASS
4. Standard Curve Data
Conc. (EU/mL) Reaction Time 1 (s) Reaction Time 2 (s) Mean (s) log(Mean) CV%
5 208 194 201 2.303 7.0% 0.5 203 223 213 2.328 9.4% 0.05 291 260 276 2.441 11.2% 0.005 1017 1004 1011 3.005 1.3%
5. Sample Endotoxin Result
Test Item Limit Replicate 1 Replicate 2 Mean Status
Bacterial Endotoxin (EU/mg) ≤ 0.50 0.022 0.056 0.039 PASS
1. Stability Study Protocol
Guideline: ICH Q1A(R2) Stability Testing, ICH Q1B Photostability
Protocol No.: STAB-SW50-2026-01
Batch No.: COA-SW50-3679
Packaging: Amber glass vial, PTFE-lined cap, N₂ backfill, secondary PE bag + desiccant
Accelerated Conditions: 40°C ± 2°C / 75% RH ± 5% RH (ICH Zone IVb)
Long-term Conditions: 25°C ± 2°C / 60% RH ± 5% RH
Photostability: ICH Option 2: ≥ 1.2 million lux·hours + ≥ 200 W·h/m² near-UV
Parameters Monitored: Appearance, Assay (HPLC), Impurities, Moisture (KF), Sterility
Timepoints: 0, 1M, 2M, 3M, 6M (accel.); 0, 3M, 6M, 9M, 12M, 18M, 24M (long-term)
2. Accelerated Stability Data (40°C / 75% RH)
Parameter Specification T=0 1 Month 2 Months 3 Months 6 Months
Appearance White powder Conforms Conforms Conforms Conforms Conforms Assay (%) 95.0 – 105.0 98.2 98.1 98.0 97.9 97.7 Max Single Impurity (%) ≤ 0.50 0.13 0.15 0.06 0.09 0.13 Total Impurities (%) ≤ 2.0 1.8 1.9 2.0 2.1 2.3 Moisture (%) ≤ 1.0 0.24 0.42 0.27 0.38 0.55
3. Long-term Stability Data (25°C / 60% RH)
Parameter Specification T=0 3 Months 6 Months 12 Months 24 Months
Appearance White powder Conforms Conforms Conforms Conforms Conforms Assay (%) 95.0 – 105.0 98.2 98.2 98.1 98.0 97.9 Max Single Impurity (%) ≤ 0.50 0.08 0.09 0.12 0.08 0.13 Moisture (%) ≤ 1.0 0.25 0.32 0.33 0.18 0.40
4. Photostability (ICH Q1B)
Parameter Before Exposure After Exposure Change Status
Appearance White lyophilized powder No significant change — PASS Assay (%) 98.2 98.1 −0.1% PASS Degradation Products (%) 1.80 1.90 +0.1% PASS
Stability Conclusion: This product remains within all specifications under both accelerated conditions (40°C/75% RH) and long-term conditions (25°C/60% RH). Recommended shelf life: 24 months when stored at -20°C, desiccated, protected from light.
GMP
Good Manufacturing Practice Certificate
Certificate of GMP Compliance
Cert. No.: CN-GMP-HN-710333-2026
This is to certify that
Hunan Aslsen Technology Co., Ltd.
Address: High-Tech Industrial Development Zone, Changsha, Hunan Province, 410205, P.R. China
Following inspection, the above facility is certified to be in compliance with the Good Manufacturing Practice for Pharmaceutical Products (2010 Revision) for the manufacture of active pharmaceutical ingredients (APIs).
Scope: Active Pharmaceutical Ingredients (Synthetic Peptides, Peptide APIs, Research-Grade Peptides, Chemical Synthesis Intermediates)
Issue Date: 2024-03-15
Valid Until: 2029-03-14
Audit Body: National Medical Products Administration (NMPA)
Audit Ref.: NMPA-GMP-7414-2024
Dr. J. Chen
Lead Auditor, NMPA
Auditor No.: NMPA-AUD-2849
ISO
SGS
QUALITY MANAGEMENT SYSTEM
ISO 9001:2015
Certified by SGS United Kingdom Ltd.
UKAS
Certificate of Registration
Certificate No.: GB24/093441
The management system of
Hunan Aslsen Technology Co., Ltd. (Homopeptide)
has been assessed and certified as meeting the requirements of ISO 9001:2015
for the following scope:
Research, development, manufacture and supply of synthetic peptides, peptide APIs, pharmaceutical raw materials, and research-grade compounds for pharmaceutical and biotech industries.
Initial Certification: 2019-06-22
Current Certification: 2025-06-22
Expiry Date: 2028-06-21
Audit Reference: SGS-QMS-175069
SGS Systems & Services
✦
Certification Body
J. Richardson
General Manager, SGS UK
UKAS Accreditation No. 014
CNAS
CHINA NATIONAL ACCREDITATION SERVICE
ISO/IEC 17025:2017
Laboratory Accreditation Certificate
Accreditation No.: CNAS L16249
Hunan Aslsen Technology Co., Ltd. — Quality Control Center
The above laboratory has been assessed by CNAS and found to comply with the requirements of ISO/IEC 17025:2017 — General requirements for the competence of testing and calibration laboratories.
Accredited Fields:
Accreditation Date: 2023-09-01
Valid Until: 2027-08-31
Dr. L. Wang
CNAS Accreditation Assessor
Assessor No.: CNAS-A2765
Certificate Verification Scan QR code or visit https://verify.homopeptide.net/COA-SW50-3679 SHA-256: ce1b0f6a70644b83... Certificate archived via blockchain — legally binding
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