Capillary hydrodynamic chromatography reveals temporal profiles of cell aggregates

Ya Ru Tang, Hsin Yi Huang, Jie Bi Hu, Rajesh Rattinam, Chun Hsien Li, Yu-Chie Chen, Pawel L. Urban*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Microbial cells are known to form aggregates. Such aggregates can be found in various matrices; for example, functional drinks. Capillary hydrodynamic chromatography (HDC) enables separation of particles by size using nanoliter-scale volumes of samples. Here we propose an approach based on HDC for characterisation of real samples containing aggregated and non-aggregated bacterial and fungal cells. Separation of cells and cell aggregates in HDC arises from the parabolic flow profile under laminar flow conditions. In the presented protocol, hydrodynamic separation is coupled with different on-line and off-line detectors (light absorption/scattering and microscopy). The method has successfully been applied in the monitoring of dynamic changes in the microbiome of probiotic drinks. Chromatographic profiles of yogurt and kefir samples obtained at different times during fermentation are in a good agreement with microscopic images. Moreover, thanks to the implementation of an area imaging detector, capillary HDC could be multiplexed and used to profile spatial gradients in cell suspensions, which arise in the course of sedimentation of cells and cell aggregates. This result shows compatibility of sedimentation analysis and capillary HDC. We believe that the approach may find applications in the profiling of functional foods and other matrices containing aggregated bioparticles.

Original languageEnglish
Pages (from-to)75-83
Number of pages9
JournalAnalytica Chimica Acta
Volume910
DOIs
StatePublished - 3 Mar 2016

Keywords

  • Aggregation
  • Cell suspension
  • Functional food
  • Hydrodynamic chromatography
  • Microbiome
  • Sedimentation analysis

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