Automated Sequential Derivatization for Gas Chromatography-[Orbitrap] Mass Spectrometry-based Metabolite Profiling of Human Blood-based Samples

• Authors: JBEBLI Akrem; COUFALÍKOVÁ Kateřina; ZANABONI Moira; BERGNA Manuela; PICENONI Renzo; KLÁNOVÁ Jana; PRICE Elliott James
• Year of publication: 2025
• Type: Article in Periodical
• Magazine / Source: Bio-protocol
• MU Faculty or unit: Faculty of Science
• web: https://bio-protocol.org/en/bpdetail?id=5196&type=0
• Doi: https://doi.org/10.21769/BioProtoc.5196
• Keywords: Automation; Derivatization; Thermo ScientificTM TriPlusTM RSH; Metabolite profiling; Gas chromatography-mass spectrometry (GC-MS)

Attached files

• 2488609.pdf

• Description: Many small molecules require derivatization to increase their volatility and to be amenable to gas chromatographic (GC) separation. Derivatization is usually time-consuming, and typical batch-wise procedures increase sample variability. Sequential automation of derivatization via robotic liquid handling enables the overlapping of sample preparation and analysis, maximizing time efficiency and minimizing variability. Herein, a protocol for the fully automated, two-stage derivatization of human blood-based samples in line with GC- [Orbitrap] mass spectrometry (MS)-based metabolomics is described. The protocol delivers a sample-to-sample runtime of 31 min, being suitable for better throughput routine metabolomic analysis.

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