Semaglutide (Fermentation) Purification Report
Primary Purification
Primary Chromatogram
Primary Purification Method
| Column | BS C18 10-120 |
| Injection | 14g/L crude |
| Detection | 210nm / 254nm |
| Mobile Phase | A: 3% HAC | B: ACN |
| Flow Rate | 60mL/min |
| Gradient | 0min:30%B → 100min:40%B |
Primary Purification Results
80.1% → 98.45%
Purity Improvement
89%
Yield
Secondary Purification
Secondary Chromatogram
Secondary Purification Method
| Column | BSC18 10-120 |
| Injection | Primary purified fraction |
| Detection | 210nm / 254nm |
| Mobile Phase A | 50mM NH4Ac + 50mM NH4Cl pH7.5 |
| Mobile Phase B | ACN |
| Flow Rate | 3mL/min |
| Gradient | 0min:25%B → 100min:40%B |
Secondary Purification Results
99.60%
Final Purity
<0.15%
Single Impurity
83.17%
Yield
Conclusion
The two-step fermentation purification process achieves exceptional semaglutide quality:
Primary Purification
- 80.1% crude → 98.45% purity (18.35% absolute improvement)
- 89% yield from crude material
- BS C18 column with HAC/ACN system
- High-flow purification (60mL/min)
Secondary Purification
- Final purity 99.60% (pharmaceutical grade)
- Single impurity <0.15% (meets strict specifications)
- 83.17% yield from primary fraction
- BSC18 column with ammonium buffer system
- Precise low-flow polishing (3mL/min)
Overall Process Performance
Combined Yield: 74.02% (89% × 83.17%)
Total Purity Gain: 80.1% → 99.6% (+19.5%)