uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Development of a capillary zone electrophoresis method to quantify E. coli L-asparaginase and its acidic variants
Univ Ghent, Dept Pharmaceut Anal, Fac Pharmaceut Sci, Drug Qual & Registrat DruQuaR Grp, Ottergemsesteenweg 460, B-9000 Ghent, Belgium.
Univ Ghent, Dept Pharmaceut Anal, Fac Pharmaceut Sci, Drug Qual & Registrat DruQuaR Grp, Ottergemsesteenweg 460, B-9000 Ghent, Belgium.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Analytical Science.ORCID iD: 0000-0001-8409-4443
Univ Ghent, Dept Pharmaceut Anal, Fac Pharmaceut Sci, Drug Qual & Registrat DruQuaR Grp, Ottergemsesteenweg 460, B-9000 Ghent, Belgium.
Show others and affiliations
2018 (English)In: Talanta: The International Journal of Pure and Applied Analytical Chemistry, ISSN 0039-9140, E-ISSN 1873-3573, Vol. 182, p. 83-91Article in journal (Refereed) Published
Abstract [en]

A capillary zone electrophoresis (CZE) method with UV detection was developed for the quantification of the E.coli L-asparaginase (L-ASNase) and its acidic variants. During the initial method development, a variety of experimental conditions were screened. Subsequently, a Design of Experiments (DoE) was used to optimize the pH and concentration of the selected background electrolyte (BGE) containing both TRIS and boric acid. Optimization was performed taking into account both the separation efficiency of L-ASNase and its acidic variants as well as overall method robustness. A repeatable separation between E.coli L-ASNase and its acidic variants was achieved on a bare fused silica capillary in combination with a BGE consisting of both 400 mM TRIS and boric acid. The method was validated for linearity, accuracy, precision, LOD, LOQ and robustness. The recovery for L-ASNase was 97.9-104.4% with a precision RSD of 1.5-3.2%, while the recovery of impurities was 92.1-109.8% with a RSD of 1.7-4.6%. The quantification limit was 1.9% (m/m). Moreover, the CZE-UV method was applied to determine the degradation rate in the presence of ammonium bicarbonate, confirming the suitability of the method. The degraded, partially charged L-ASNase was evaluated for its in-vitro enzymatic activity showing an insignificant different enzyme activity compared to the unmodified sample.

Place, publisher, year, edition, pages
ELSEVIER SCIENCE BV , 2018. Vol. 182, p. 83-91
Keywords [en]
l-asparaginase, Capillary zone electrophoresis, Acidic impurities, Design of experiments (DoE)
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-353099DOI: 10.1016/j.talanta.2018.01.048ISI: 000428229200010PubMedID: 29501203OAI: oai:DiVA.org:uu-353099DiVA, id: diva2:1221576
Available from: 2018-06-20 Created: 2018-06-20 Last updated: 2019-10-18Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textPubMed

Authority records BETA

Sänger - van de Griend, Cari

Search in DiVA

By author/editor
Sänger - van de Griend, CariHaselberg, Rob
By organisation
Analytical Science
In the same journal
Talanta: The International Journal of Pure and Applied Analytical Chemistry
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 180 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf