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Investigation of robustness for supercritical fluid chromatography separation of peptides: Isocratic vs gradient mode
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Farmaceutiska fakulteten, Institutionen för läkemedelskemi, Farmakognosi. Karlstad Univ, Dept Engn & Chem Sci, SE-65188 Karlstad, Sweden.
Karlstad Univ, Dept Engn & Chem Sci, SE-65188 Karlstad, Sweden.
Karlstad Univ, Dept Engn & Chem Sci, SE-65188 Karlstad, Sweden;Rzeszow Univ Technol, Dept Chem & Proc Engn, PL-35959 Rzeszow, Poland.
AstraZeneca, IMED Biotech Unit, Early Product Dev, Pharmaceut Sci, Gothenburg, Sweden.
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2018 (Engelska)Ingår i: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1568, s. 177-187Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

We investigated and compared the robustness of supercritical fluid chromatography (SFC) separations of the peptide gramicidin, using either isocratic or gradient elution. This was done using design of experiments in a design space of co-solvent fraction, water mass fraction in co-solvent, pressure, and temperature. The density of the eluent (CO2-MeOH-H2O) was experimentally determined using a Coriolis mass flow meter to calculate the volumetric flow rate required by the design. For both retention models, the most important factor was the total co-solvent fraction and water mass fraction in co-solvent. Comparing the elution modes, we found that gradient elution was more than three times more robust than isocratic elution. We also observed a relationship between the sensitivity to changes and the gradient steepness and used this to draw general conclusions beyond the studied experimental system. To test the robustness in a practical context, both the isocratic and gradient separations were transferred to another laboratory. The gradient elution was highly reproducible between laboratories, whereas the isocratic system was not. Using measurements of the actual operational conditions (not the set system conditions), the isocratic deviation was quantitatively explained using the retention model. The findings indicate the benefits of using gradient elution in SFC as well as the importance of measuring the actual operational conditions to be able to explain observed differences between laboratories when conducting method transfer.

Ort, förlag, år, upplaga, sidor
ELSEVIER SCIENCE BV , 2018. Vol. 1568, s. 177-187
Nyckelord [en]
SFC, Peptide, Gramicidin, Robustness, Method transfer, Water
Nationell ämneskategori
Analytisk kemi
Identifikatorer
URN: urn:nbn:se:uu:diva-364233DOI: 10.1016/j.chroma.2018.07.029ISI: 000443669600019PubMedID: 30072233OAI: oai:DiVA.org:uu-364233DiVA, id: diva2:1258458
Forskningsfinansiär
Vetenskapsrådet, 2015-04627ÅForsk (Ångpanneföreningens Forskningsstiftelse), 17/500Tillgänglig från: 2018-10-24 Skapad: 2018-10-24 Senast uppdaterad: 2018-10-24Bibliografiskt granskad

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Enmark, Martin

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Enmark, MartinSamuelsson, JorgenFornstedt, Torgny
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