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- Title
Ternary eluent compositions in supercritical fluid chromatography improved fingerprinting of therapeutic peptides.
- Authors
Neumann, Jonas; Schmidtsdorff, Sebastian; Schmidt, Alexander H.; Parr, Maria K.
- Abstract
Currently, little information has been published on the application of ternary eluent compositions in supercritical fluid chromatography for separating peptides. This work investigates the benefits of adding acetonitrile to methanol as the modifier. Three cyclic antibiotic peptides (bacitracin, colistin, and daptomycin) ranging between 1000 and 2000 Da were chosen as model substances. The ternary mixture of carbon dioxide, methanol, and acetonitrile is optimized to increase the resolution of the peptide's fingerprint. In addition, varying compositions of methanol and acetonitrile were found to change the elution order of the analytes, which is a valuable tool during method development. An individual gradient method using two Torus 2‐PIC columns (each 100 × 3.0 mm, 1.7 μm), carbon dioxide, and a modifier consisting of acetonitrile/methanol/water/methanesulfonic acid (60:40:2:0.1, v:v:v:v) was optimized for each of the peptides. Subsequently, a generic method development protocol applicable to polypeptides is proposed.
- Subjects
CYCLIC peptides; PEPTIDES; SUPERCRITICAL fluid chromatography; PEPTIDE mass fingerprinting; CARBON dioxide; POLYPEPTIDES; COLISTIN
- Publication
Journal of Separation Science, 2023, Vol 46, Issue 5, p1
- ISSN
1615-9306
- Publication type
Article
- DOI
10.1002/jssc.202201007