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. 2020 Oct 14;5(42):27050-27056.
doi: 10.1021/acsomega.0c02135. eCollection 2020 Oct 27.

Terbium Excitation Spectroscopy as a Detection Method for Chromatographic Separation of Lanthanide-Binding Biomolecules

Wojciech Jurkowski  1 , Marcus Heilmann  2 , Anna M Becker  3 , Rainer Buchholz  3 , Thomas B Brück  1
Affiliations

Terbium Excitation Spectroscopy as a Detection Method for Chromatographic Separation of Lanthanide-Binding Biomolecules

Wojciech Jurkowski et al. ACS Omega. .

Abstract

Studies of biosorption and bioaccumulation of heavy metals deal mostly with challenging, inhomogeneous, and complex materials. Therefore, most reports describe only application studies, while fundamental research is limited to indirect methods and speculations on the binding mechanisms. In this study, we describe a method for detecting and isolating heavy metal-binding biomolecules directly from crude extracts. The underlying principle is terbium sensitization and fluorescence excitation spectroscopy used offline after a chromatographic run. Compounds interacting with metal ions inevitably change the coordination sphere of terbium, which is reflected in the excitation spectrum leading to metal-specific luminescence. Main advantages of our approach include simple, fast, and inexpensive experiment design, nondestructive measurements, and detection limits far below 1 mg. Here, we have applied our method for three promising biosorbents (green algae, moss, and cyanobacterium) and obtained first information on the character of active compounds isolated from each species.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
General workflow of the method.
Figure 2
Figure 2
Chromatogram from raw extract of C. brevissima with fractional spectroscopy separated on the SEC column (Superdex-75 10/300 GL). Injection volume: 500 μL, F = 0.5 mL/min, 10 °C. UV detection—absorbance at 190, 230, 270, and 400 nm. Spectra from left: absorbance in the UV region, background excitation (1) without terbium, excitation (2) with added terbium, background subtracted excitation with terbium. All excitation spectra collected at 545 nm emission using the dual monochromator with xenon arc lamp source.
Figure 3
Figure 3
Luminescence excitation for 545 nm (solid line) and emission spectra for 264 nm (dashed line). Fraction 17 refers to deionized water used as a reference for the Tb signal.
Figure 4
Figure 4
Luminescence excitation spectra for 545 nm for complexes of terbium with different metal chelators.
See this image and copyright information in PMC

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