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Vidjay Christinat.pdf
Bachelor thesis

Investigation of Kdo derivates in endotoxin standards

  • Sion : Haute Ecole d'Ingénierie, HES-SO Valais-Wallis, 2022

Bachelor of Science (BSc) HES-SO ingénieur·e en Technologies du vivant: Haute Ecole d'Ingénierie, HES-SO Valais-Wallis, 2022

English Endotoxins (ETs) are frequent and dangerous contaminants in pharmaceutical and biotechnological products. They originate from the surface of Gram-negative bacteria where they are non-covalently attached. ETs cause severe health issues as fever, organ failures or deaths in humans when they enter the blood stream.
Today ET analysis is mainly done with biological assays as the rabbit pyrogen test (RPT), the monocyte activation test (MAT) or the Limulus Amoebocyte Lysate (LAL) assay. All these tests show serious setbacks e.g., they either do not convey to animal-welfare guidelines or they suffer from severe matrix effects commonly known as low endotoxin recovery (LER). That can lead to a 100 % masking of ETs in pharmaceutical preparations and with that give false negative test results what can endanger human health.
At HES-SO Valais-Wallis, a chemical quantitative ET assay (KDO-DMB-LC assay) was developed that overcomes the substantial drawbacks of the today commonly used tests. It is based on an ET specific sugar acid (KDO). One to four KDOs are present in the inner core of each ET molecule. In the KDO-DMB-LC assay KDO is liberated by mild acidic hydrolysis and subsequently derivatized with a fluorescent dye (DMB), allowing its sensitive detection. After LC separation, the KDO-DMB analyte is detected via a fluorescence detector.
The ET specific sugar acid KDO can bear several functional groups such as pyrophosphates, phosphates or phosphoethanolamines. Different ET types show different modification patterns. The different, until now known KDO derivates are separated in the current isocratic KDO-DMB assay. Until now only the free, not functionalized KDO is used to calculate the ET content of a sample.
Language
  • English
Classification
Analytical chemistry
Notes
  • Haute Ecole d'Ingénierie Valais
  • Technologies du vivant - Life Technologies
  • Chimie analytique
  • hesso:heivs
Persistent URL
https://sonar.rero.ch/hesso/documents/324726
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