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Capillary electrophoresis with preparative isoelectric focusing preconcentration for sensitive determination of amphotericin B in human blood serum
J Salplachta, M Horka, K Slais
Jazyk angličtina Země Nizozemsko
Grantová podpora
NV16-29916A
MZ0
CEP - Centrální evidence projektů
- MeSH
- amfotericin B * izolace a purifikace krev MeSH
- biochemická analýza krve * metody MeSH
- elektroforéza kapilární * metody MeSH
- isoelektrická fokusace MeSH
- lidé MeSH
- limita detekce * MeSH
- Check Tag
- lidé MeSH
Amphotericin B (AmB) is still, despite its severe nephrotoxicity, the first-line agent in the management of serious systemic fungal infections. A sensitive and reliable method is therefore required to control AmB concentration in body fluids of a patient. This study demonstrates the potential of the off-line combination of preparative isoelectric focusing (IEF) with capillary isoelectric focusing (CIEF) or capillary zone electrophoresis (CZE) in the determination of AmB in human blood serum. The required value of the isoelectric point of AmB was determined to be 6.1 using the CIEF technique. Preparative IEF served as a pre-separation and concentration technique. The pH gradient was traced by colored low molecular pI markers. The collected fraction with AmB was easily processed and then analyzed by CIEF and CZE. Tens of picograms of AmB in human blood serum sample can be determined by a combination of preparative IEF with CZE. The method was linear in the AmB concentration range of 0.3-600ngmL
Citace poskytuje Crossref.org
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- $a Capillary electrophoresis with preparative isoelectric focusing preconcentration for sensitive determination of amphotericin B in human blood serum / $c J Salplachta, M Horka, K Slais
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- $a Amphotericin B (AmB) is still, despite its severe nephrotoxicity, the first-line agent in the management of serious systemic fungal infections. A sensitive and reliable method is therefore required to control AmB concentration in body fluids of a patient. This study demonstrates the potential of the off-line combination of preparative isoelectric focusing (IEF) with capillary isoelectric focusing (CIEF) or capillary zone electrophoresis (CZE) in the determination of AmB in human blood serum. The required value of the isoelectric point of AmB was determined to be 6.1 using the CIEF technique. Preparative IEF served as a pre-separation and concentration technique. The pH gradient was traced by colored low molecular pI markers. The collected fraction with AmB was easily processed and then analyzed by CIEF and CZE. Tens of picograms of AmB in human blood serum sample can be determined by a combination of preparative IEF with CZE. The method was linear in the AmB concentration range of 0.3-600ngmL<ovid:sup>-1</ovid:sup>. The recovery ranged from 93% to 98%.<ovid:br/><ovid:br/> Copyright © 2018 Elsevier B.V. All rights reserved.
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