Journalartikel

Jahr der Veröffentlichung: 2022

Zeitschrift: Journal of Chromatography A

Bandnummer: 1679

ISSN: 0021-9673

eISSN: 1873-3778

DOI Link: https://doi.org/10.1016/j.chroma.2022.463393

Verlag: Elsevier

Abstract: 

The European Union has banned the use of antibiotic growth promoters in animal production, which has led to increased use of probiotic microorganisms. These feed additives result in higher costs for farmers, which is why the demand for a quality control system to quantify probiotics in feeds has increased in recent years. Imaging high-performance thin-layer chromatography (HPTLC) was proven to be a ro-bust method for determining the probiotic Bacillus subtilis DSM 29784 strain based on the production of selective bacterial metabolites and thus characteristic metabolite pattern. However, to quantify the spe-cific probiotic strain in the feed, identification of a strain-specific metabolite not produced by genetically very similar bacteria is necessary. Compared to five bacteria with high genetic similarity, a strain-specific metabolite was formed in the probiotic bacteria by a two-step cultivation procedure. Among others, an-timicrobial properties were found for this metabolite, which indicated probiotic activity. The hyphenation of normal-phase HPTLC with reversed-phase high-performance liquid chromatography diode array detec-tion and high-resolution mass spectrometry allowed the preliminary assignment of this strain-specific metabolite to the molecular formula C35H44N6O2 (580.3527 Da). This metabolite, produced each time via an upstream cultivation process to generate the standard levels, was used for the quantification of pro-biotic active cells in the feed. Data on selectivity, linearity, detection limit, recovery, and precision have shown the good performance of the method. 

Harvard-Zitierstil: Kruse, S., Becker, S., Pierre, F. and Morlock, G. (2022) Strain-specific quantification of probiotic Bacillus subtilis in feed by imaging high-performance thin-layer chromatography, Journal of Chromatography A, 1679, Article 463393. https://doi.org/10.1016/j.chroma.2022.463393

APA-Zitierstil: Kruse, S., Becker, S., Pierre, F., & Morlock, G. (2022). Strain-specific quantification of probiotic Bacillus subtilis in feed by imaging high-performance thin-layer chromatography. Journal of Chromatography A. 1679, Article 463393. https://doi.org/10.1016/j.chroma.2022.463393