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Retatrutide Purification

Retatrutide Purification Report

BBN-GLP1-RET-001 March 2023

 Primary Purification

Primary Chromatogram

Primary Purification Method

Column BS C18 10μm 120Å
Injection 14g/L crude
Detection 210nm / 254nm
Mobile Phase A: 3% HAC | B: Preparative ACN
Flow Rate 3mL/min
Gradient 0min:30%B → 100min:45%B

Primary Purification Results

82% → 98.54%
Purity Improvement
86.85%
Yield
0.5%
Max Single Impurity

 Secondary Purification

Secondary Chromatogram

Secondary Purification Method

Column BS C18 10μm 120Å
Injection Primary purified fraction
Detection 280nm / 254nm
Mobile Phase A 50mM NH4Ac + 50mM NH4Cl pH6.5
Mobile Phase B Preparative ACN
Flow Rate 3mL/min
Gradient 0min:30%B → 100min:40%B

Secondary Purification Results

99.5%
Final Purity
0.25%
Max Single Impurity
89.9%
Yield

Conclusion

The two-step purification process achieves exceptional retatrutide quality:

Primary Purification

  • 82% crude → 98.54% purity (+16.54% absolute improvement)
  • 86.85% yield from crude material
  • Maximum single impurity: 0.5%
  • BS C18 column with HAC/ACN system
  • 210nm primary detection wavelength

Secondary Purification

  • Final purity 99.5% (ultra-pure grade)
  • Maximum single impurity: 0.25% (50% reduction)
  • 89.9% yield from primary fraction
  • Ammonium acetate/chloride buffer system (pH6.5)
  • 280nm primary detection for final QC

Overall Process Performance

Combined Yield: 78.08% (86.85% × 89.9%)
Total Purity Gain: 82% → 99.5% (+17.5%)
Impurity Reduction: From 18% → 0.5% → 0.25%

Note: The BS C18 column demonstrates excellent performance for both purification steps, with buffer system modification and wavelength optimization for final polishing. The process delivers ultra-pure retatrutide suitable for pharmaceutical applications.

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