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Investigation of robustness for supercritical fluid chromatography separation of peptides: Isocratic vs gradient mode
Karlstads universitet, Fakulteten för hälsa, natur- och teknikvetenskap (from 2013), Institutionen för ingenjörsvetenskap och fysik (from 2013).ORCID-id: 0000-0002-8943-6286
Karlstads universitet, Fakulteten för hälsa, natur- och teknikvetenskap (from 2013), Institutionen för ingenjörs- och kemivetenskaper (from 2013).
Rzeszów University of Technology, Poland.
AstraZeneca.
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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, 2018. Vol. 1568, s. 177-187
Nyckelord [en]
Gramicidin, Method transfer, Peptide, Robustness, SFC, Water, Design of experiments, Effluent treatment, Gas chromatography, Oceanography, Peptides, Robustness (control systems), Supercritical fluids, Coriolis mass flow meter, Experimental system, Gradient separations, Method transfers, Operational conditions, Super critical fluid chromatography, Volumetric flow rate, Solvents
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URN: urn:nbn:se:kau:diva-69063DOI: 10.1016/j.chroma.2018.07.029ISI: 000443669600019OAI: oai:DiVA.org:kau-69063DiVA, id: diva2:1245843
Tillgänglig från: 2018-09-06 Skapad: 2018-09-06 Senast uppdaterad: 2018-11-22Bibliografiskt granskad

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Enmark, MartinGlenne, EmelieSamuelsson, JörgenFornstedt, Torgny

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Enmark, MartinGlenne, EmelieSamuelsson, JörgenFornstedt, Torgny
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Institutionen för ingenjörsvetenskap och fysik (from 2013)Institutionen för ingenjörs- och kemivetenskaper (from 2013)
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